WO2019170059A1 - Procédé d'enrobage d'un médicament dans une ferritine et produit associé - Google Patents

Procédé d'enrobage d'un médicament dans une ferritine et produit associé Download PDF

Info

Publication number
WO2019170059A1
WO2019170059A1 PCT/CN2019/076895 CN2019076895W WO2019170059A1 WO 2019170059 A1 WO2019170059 A1 WO 2019170059A1 CN 2019076895 W CN2019076895 W CN 2019076895W WO 2019170059 A1 WO2019170059 A1 WO 2019170059A1
Authority
WO
WIPO (PCT)
Prior art keywords
ferritin
drug
guanidine hydrochloride
case
hfn
Prior art date
Application number
PCT/CN2019/076895
Other languages
English (en)
Chinese (zh)
Inventor
阎锡蕴
范克龙
朱京东
Original Assignee
昆山新蕴达生物科技有限公司
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 昆山新蕴达生物科技有限公司 filed Critical 昆山新蕴达生物科技有限公司
Publication of WO2019170059A1 publication Critical patent/WO2019170059A1/fr

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
    • 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
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • 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
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/20—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing sulfur, e.g. dimethyl sulfoxide [DMSO], docusate, sodium lauryl sulfate or aminosulfonic acids
    • 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
    • A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • 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
    • A61K47/6949—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 inclusion complexes, e.g. clathrates, cavitates or fullerenes
    • 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
    • A61K9/5123—Organic compounds, e.g. fats, sugars
    • 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
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals

Definitions

  • the present invention relates to a method of ferritin-loaded drug and a product thereof.
  • the present invention relates to methods and products for depolymerization using SDS or guanidine hydrochloride or urea, renatured ferritin such as whole human ferritin, and drugs such as doxorubicin.
  • Ferritin is an important functional protein involved in and maintain iron metabolism balance. It is a kind of protein containing high iron content widely distributed in animals, plants and microbial cells. From bacteria to humans, although the ferritin amino acid sequences of different organisms are greatly different, their structures are similar.
  • the typical ferritin structure is composed of a protein shell and an iron core.
  • the protein shell is a cage structure formed by self-assembly of 24 subunits (outer diameter 12 nm, inner diameter 8 nm).
  • the main component of the iron core is ferrihydrite. (5Fe 2 O 3 ⁇ 9H 2 O).
  • the ferritin shell is usually composed of two protein subunits (H and L). In different tissues and organs of the body, the proportion of H and L subunits in the ferritin molecule is different.
  • Human H ferritin refers to ferritin formed by self-assembly of the H subunit of human iron eggs.
  • the H subunit of human ferritin can self-assemble to form a cage protein, and the number of H subunits of human ferritin is usually 24.
  • the full length amino acid sequence of human H ferritin is shown in SEQ ID NO: 1.
  • Human L ferritin refers to ferritin formed by self-assembly of the L subunit of human iron eggs.
  • the L subunit of human ferritin can self-assemble to form a cage protein, and the number of L subunits of human ferritin is usually 24.
  • the full length amino acid sequence of human L ferritin is shown in SEQ ID NO: 2.
  • Pyrococcus furiosus ferritin is a ferritin formed by self-assembly of the fibrinogen ferritin subunit.
  • the fibrinogen ferritin ferritin subunit can self-assemble to form a cage protein, and the number of subunits of the human Pyrococcus ferulil ferritin is usually 24.
  • the full-length amino acid sequence of Pyrococcus furiosus ferritin is shown in SEQ ID NO: 3.
  • the drug loading of ferritin from different sources depends mainly on two ways: 1.
  • the drug enters the inside of the protein shell through ion channels or hydrophobic channels on ferritin under specific treatment conditions; 2.
  • a high concentration of urea the protein shell is depolymerized, and the drug to be entrapped is added during the subsequent recombination process, thereby realizing the loading of the drug inside the protein shell.
  • HFn entrapped doxorubicin is particularly efficient under high-concentration urea conditions, and 33 molecules of doxorubicin are loaded per molecule of HFn; the brief procedure of the method is: adding human HFn to a final concentration greater than 6M In the urea, react at room temperature for 30 minutes, then add appropriate amount of doxorubicin reagent, and protect from light for 10 minutes, then use dialysis method to gradually reduce the urea concentration in the reaction system to 0, thus achieving doxorubicin Envelope in HFn.
  • drugs that need to be entrapped in the prior art. These drugs can either avoid toxic side effects after encapsulation, or avoid low solubility to improve utilization, or avoid degradation, and so on.
  • the poorly soluble drug docetaxel can avoid the determination that its solubility is low and difficult to apply after encapsulation.
  • Doxorubicin is an anti-tumor antibiotic that inhibits the synthesis of RNA and DNA. It has the strongest inhibitory effect on RNA, has a broad anti-tumor spectrum, and has a role in various tumors. It is a non-specific drug for various growth. Periodic tumor cells have a killing effect. It is mainly used for acute leukemia and is effective for acute lymphoblastic leukemia and granulocyte leukemia. It is generally used as a second-line drug, that is, it can be considered when the drug is the drug of choice. For malignant lymphoma, it can be used as the drug of choice for alternate use. It has a certain effect on breast cancer, sarcoma, lung cancer, bladder cancer and other various cancers, and is often used in combination with other anticancer drugs.
  • doxorubicin when used directly in the injection, doxorubicin has a wide range of biochemical effects on the body due to its diffuse distribution throughout the blood, and has a strong cytotoxic effect.
  • the main toxic reactions are: white blood cells and thrombocytopenia, about 60% to 80% of patients can occur; 100% of patients have varying degrees of hair loss, can resume growth after stopping the drug; cardiotoxicity, manifested as arrhythmia, ST- T changes, often appear 1 to 6 months after withdrawal; nausea, loss of appetite; drug spillage outside the blood vessels can cause tissue ulcers and necrosis.
  • urine can appear red after administration.
  • a first aspect of the invention relates to a method of ferritin-loaded medicament comprising the steps of:
  • the denaturing agent is a substance which can weaken or break the hydrogen bond interaction between the ferritin subunits, thereby depolymerizing the ferritin.
  • the denaturing agent means an anionic surfactant or guanidine hydrochloride or urea, preferably an anion.
  • the surfactant is selected from the group consisting of SDS, stearic acid and/or sodium dodecylbenzene sulfonate; wherein a certain concentration in the case of an anionic surfactant means a weight ratio of about 1% to 10%, in the case of guanidine hydrochloride In the case of 4M to 8M, in the case of urea, it is 3 to 10M; in the case of anionic surfactant, it means 10 to 120 minutes, and in the case of guanidine hydrochloride, it is 30 to 300 minutes.
  • a certain temperature means 30 ° C to 100 ° C in the case of an anionic surfactant, 25 ° C to 70 ° C in the case of guanidine hydrochloride, and 15 in the case of urea. °C ⁇ 75 °C.
  • the ferritin is selected from the group consisting of natural ferritin or recombinant ferritin that can form a cage structure and variants thereof, preferably, ferritin is a fully human ferritin, and more preferably, ferritin is a full human heavy chain Ferritin.
  • the denaturant is selected from the group consisting of SDS or guanidine hydrochloride.
  • the drug is selected from an antitumor drug or a non-antitumor drug, preferably, the antitumor drug is selected from the group consisting of a tumor antibiotic, a natural source antineoplastic agent, a metal compound, a radioisotope, an alkylating agent, an antimetabolite resistant
  • the oncology drug, the hormone antitumor drug preferably, the drug is selected from the group consisting of a tumor antibiotic, a natural source antitumor drug, a radioisotope or an alkylating agent, and more preferably, the drug is selected from a tumor antibiotic, and more preferably, the drug is selected from the group consisting of Neomycin, daunorubicin, choline, curcumin, nimustine, carmustine, lomustine, cyclophosphamide, ifosfamide, glycophosphorus, fluoroferron, doxine Acridine, 5-fluorouracil, guanidine, thioguan
  • the agonist drug is selected from the group consisting of muscarinic, pilocarpine, gamma Inhibitor is selected from bis-fluoro ketones, the antioxidant is selected from melatonin, anesthetic selected from anthracene amine.
  • the denaturant in the removal solution is carried out by desalting, preferably using a desalting column.
  • the mass ratio of ferritin-incorporated drug in the case of an anionic surfactant is from 5% to 60%, preferably at least 8%, 10%, 15%, 20%, 25%, 30 %, 35%, 40%, 45%, 50%, 55%, 60% or higher, more preferably at least 20%, 25%, 30% or 35%; ferritin in the case of a guanidine hydrochloride denaturing agent
  • the amount of the entrapped drug is 5%-60%, preferably at least 8%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60% % or higher, more preferably, at least 20%, 25%, 30% or 35%; in the case of a urea denaturant, the amount of ferritin-encapsulated drug is 5% - 60%, preferably at least 8%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60% or higher, more preferably at least 20%, 25%, 30% or 35%.
  • the ferritin content of the solution ranges from 0.1 to 100 mg/ml, preferably from 0.2 to 10 mg/ml, more preferably from 0.3 to 5 mg/ml, more preferably from 0.5 to 2 mg/ml, more preferably , 0.7 to 1.4 mg/ml; and/or, the content of the drug to be contained is in the range of 0.1 to 50 mg/ml, preferably 0.2 to 10 mg/ml, more preferably 0.3 to 5 mg/ml, more preferably 0.5 to 2 mg/ml, more preferably 0.7 to 1.4 mg/ml.
  • a certain concentration refers to a weight ratio of about 5% to 10% in the case of an anionic surfactant, 6M to 8M in the case of guanidine hydrochloride, and 5 to 5 in the case of guanidine hydrochloride.
  • anionic surfactant means 10 ⁇ 30min
  • guanidine hydrochloride means 30min ⁇ 120min
  • urea means 20 ⁇ 200min
  • a certain temperature in anionic surfactant In the case of 65 ° C to 75 ° C, in the case of guanidine hydrochloride, it means 40 ° C to 60 ° C, and in the case of urea, it is 20 ° C to 50 ° C.
  • a certain concentration refers to a weight ratio of about 6.5% to 8% in the case of an anionic surfactant, and 6.5M to 7.5M in the case of hydrazine hydrochloride, and in the case of hydrazine hydrochloride.
  • a certain time in the case of anionic surfactant means 10-20min, in the case of guanidine hydrochloride means 50min ⁇ 72min, in the case of guanidine hydrochloride means 25 ⁇ 100min; a certain temperature in anion In the case of a surfactant, it is 68 to 72 ° C, in the case of barium hydrochloride, it is 45 to 55 ° C, and in the case of barium hydrochloride, it is 22 to 35 ° C.
  • the solution contains an additive that promotes dissolution of the drug and/or aggregation of ferritin, preferably selected from DMA, DMF, DMSO, or mixtures thereof.
  • the mass ratio of ferritin to the entrapped drug in the solution is from 5:1 to 1:5.
  • the buffer system of the solution is Tris-HCl buffer, phosphate buffer (PBS), carbonate buffer, glycine buffer or citrate buffer, and the pH of the buffer is between 3 and 10.
  • the concentration of the buffer is between 20 and 500 mM.
  • a second aspect of the invention relates to a drug-loaded ferritin obtained by the method of ferritin-incorporated drug according to the above first aspect.
  • the method for encapsulating a drug with ferritin of the invention can greatly increase the drug entrapment amount, and the clinical administration dose will be greatly reduced under the premise of achieving the same therapeutic effect.
  • FIG. 1 Chromatographic peak position of HFn undepolymerized after treatment with 4M guanidine hydrochloride at 25 ° C for 60 min (mobile phase containing 4 M guanidine hydrochloride).
  • FIG. 20 Chromatographic peak position of HFn undepolymerized after treatment with 4M guanidine hydrochloride at 25 ° C for 120 min (mobile phase containing 4 M guanidine hydrochloride).
  • FIG. 21 Chromatographic peak position of HFn 14.2% depolymerization after treatment with 5.5 M guanidine hydrochloride at 25 ° C for 60 min (mobile phase containing 5.5 M guanidine hydrochloride).
  • FIG. 22 Chromatographic peak position of HFn 17.2% depolymerization after treatment with 5.5 M guanidine hydrochloride at 25 ° C for 90 min (mobile phase containing 5.5 M guanidine hydrochloride).
  • FIG. 23 Chromatographic peak position of 18.7% depolymerization of HFn after treatment with 5.5 M guanidine hydrochloride at 25 ° C for 120 min (mobile phase containing 5.5 M guanidine hydrochloride).
  • FIG. 24 Chromatographic peak position of 24.6% depolymerization of HFn after treatment with 6M guanidine hydrochloride at 25 ° C for 60 min (mobile phase containing 6 M guanidine hydrochloride).
  • FIG. 25 Chromatographic peak position of HFn 33.8% depolymerization after treatment with 6M guanidine hydrochloride at 25 ° C for 90 min (mobile phase containing 6 M guanidine hydrochloride).
  • FIG. 26 Chromatographic peak position of 34.3% depolymerization of HFn after treatment with 6M guanidine hydrochloride at 25 ° C for 120 min (mobile phase containing 6 M guanidine hydrochloride).
  • FIG. 27 Chromatographic peak position of 6.5 n% depolymerization of 6.5 n HCl hydrochloride at 25 ° C for 60 min (mobile phase containing 6.5 M guanidine hydrochloride).
  • FIG. 28 Chromatographic peak position of 6.5 nm depolymerization of HFn after treatment with 6.5 M guanidine hydrochloride at 25 ° C for 90 min (mobile phase containing 6.5 M guanidine hydrochloride).
  • FIG. 29 Chromatographic peak position of 4.6 degN 2.44% depolymerization after 6.5 M guanidine hydrochloride, 25 ° C, 120 min treatment (mobile phase containing 6.5 M guanidine hydrochloride).
  • FIG. 30 Chromatographic peak position of 53.2% depolymerization of HFn after treatment with 7M guanidine hydrochloride at 25 ° C for 60 min (mobile phase containing 7 M guanidine hydrochloride).
  • FIG. 31 Chromatographic peak position of 57.9% depolymerization of HFn after treatment with 7M guanidine hydrochloride at 25 ° C for 90 min (mobile phase containing 7 M guanidine hydrochloride).
  • FIG. 32 Chromatographic peak position of 60.9% depolymerization of HFn after treatment with 7M guanidine hydrochloride at 25 ° C for 120 min (mobile phase containing 7 M guanidine hydrochloride).
  • FIG. 33 Chromatographic peak position of 63.7% depolymerization of HFn after treatment with 7M guanidine hydrochloride at 25 ° C for 150 min (mobile phase containing 7 M guanidine hydrochloride).
  • FIG. 34 Chromatographic peak position of 65.8% depolymerization of HFn after treatment with 7M guanidine hydrochloride at 25 ° C for 180 min (mobile phase containing 7 M guanidine hydrochloride).
  • FIG. 35 Chromatographic peak position of 67.9% depolymerization of HFn after treatment with 7M guanidine hydrochloride at 25 ° C for 210 min (mobile phase containing 7 M guanidine hydrochloride).
  • FIG. 36 Chromatographic peak position of HFN 66% depolymerization after treatment with 7.5 M guanidine hydrochloride at 25 ° C for 60 min (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG. 37 Chromatographic peak position of 74.7% depolymerization of HFn after treatment with 7.5 M guanidine hydrochloride at 25 ° C for 90 min (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG. 38 Chromatographic peak position of 79.8% depolymerization of HFn after treatment with 7.5 M guanidine hydrochloride at 25 ° C for 120 min (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG 39 Chromatographic peak position of HFn 86.1% depolymerization after treatment with 7.5 M guanidine hydrochloride at 25 ° C for 150 min (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG. 40 Chromatographic peak position of HFN 88.9% depolymerization after treatment with 7.5 M guanidine hydrochloride at 25 ° C for 180 min (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG 41 Chromatographic peak position of 95.8% depolymerization of HFn after treatment with 7.5 M guanidine hydrochloride at 25 ° C for 210 min (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG. 42 Chromatographic peak position of 96.6% depolymerization of HFn after treatment with 7.5 M guanidine hydrochloride at 25 ° C for 240 min (mobile phase containing 7.5 M guanidine hydrochloride).
  • Figure 43 Peak position of the 7.4 depolymerization of HFn 97.4% after 7.5 M guanidine hydrochloride, 25 ° C, 270 min treatment (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG 44 Chromatographic peak position of HFn 100% depolymerization after treatment with 7.5 M guanidine hydrochloride at 25 ° C for 300 min (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG. 45 Chromatographic peak position of HFn 100% depolymerization after treatment with 7.5 M guanidine hydrochloride at 25 ° C for 330 min (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG. 46 Chromatographic peak position of HFn 100% depolymerization after 7.5 M guanidine hydrochloride, 25 ° C, 360 min treatment (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG. 47 Chromatographic peak position of 93.9% depolymerization of HFn after treatment with 8M guanidine hydrochloride at 25 ° C for 60 min (mobile phase containing 8 M guanidine hydrochloride).
  • FIG. 48 Chromatographic peak position of 98.1% depolymerization of HFn after treatment with 8M guanidine hydrochloride at 25 ° C for 90 min (mobile phase containing 8 M guanidine hydrochloride).
  • FIG. 49 Chromatographic peak position of 98.6% depolymerization of HFn after treatment with 8M guanidine hydrochloride at 25 ° C for 120 min (mobile phase containing 8 M guanidine hydrochloride).
  • Figure 50 Chromatographic peak position of HFn 100% depolymerization after treatment with 8 M guanidine hydrochloride at 25 ° C for 150 min (mobile phase containing 8 M guanidine hydrochloride).
  • Figure 51 Chromatographic peak position of 39.5% depolymerization of HFn after treatment with 4M guanidine hydrochloride at 70 ° C for 30 min (mobile phase containing 4 M guanidine hydrochloride).
  • FIG. 52 Chromatographic peak position of HFn 53.3% depolymerization after treatment with 4M guanidine hydrochloride at 70 ° C for 60 min (mobile phase containing 4 M guanidine hydrochloride).
  • FIG. 53 Chromatographic peak position of 82.6% depolymerization of HFn after treatment with 4M guanidine hydrochloride at 70 ° C for 120 min (mobile phase containing 4 M guanidine hydrochloride).
  • FIG. 54 Chromatographic peak position of 91.8% depolymerization of HFn after treatment with 4M guanidine hydrochloride at 70 ° C for 180 min (mobile phase containing 4 M guanidine hydrochloride).
  • FIG. 55 Chromatographic peak position of 64.1% depolymerization of HFn after treatment with 5M guanidine hydrochloride at 70 ° C for 30 min (mobile phase containing 5 M guanidine hydrochloride).
  • FIG. 56 Chromatographic peak position of 92.7% depolymerization of HFn after treatment with 5M guanidine hydrochloride at 70 ° C for 60 min (mobile phase containing 5 M guanidine hydrochloride).
  • FIG. 57 Chromatographic peak position of HFn 100% depolymerization after treatment with 5M guanidine hydrochloride at 70 ° C for 120 min (mobile phase containing 5 M guanidine hydrochloride).
  • Figure 58 Chromatographic peak position of 41.5% depolymerization of HFn after treatment with 6M guanidine hydrochloride at 37 ° C for 120 min (mobile phase containing 6 M guanidine hydrochloride).
  • FIG. 59 Chromatographic peak position of 63.45% depolymerization of HFn after treatment with 6M guanidine hydrochloride at 50 ° C for 30 min (mobile phase containing 6 M guanidine hydrochloride).
  • Figure 60 Chromatographic peak position of 44.6% depolymerization of HFn after treatment with 6M guanidine hydrochloride at 50 ° C for 60 min (mobile phase containing 6 M guanidine hydrochloride).
  • FIG. 61 Chromatographic peak position of HFn 61.2% depolymerization after treatment with 6M guanidine hydrochloride at 50 ° C for 120 min (mobile phase containing 6 M guanidine hydrochloride).
  • FIG. 62 Chromatographic peak position of 67.8% depolymerization of HFn after treatment with 6M guanidine hydrochloride at 50 ° C for 180 min (mobile phase containing 6 M guanidine hydrochloride).
  • FIG. 63 Chromatographic peak position of 76.6% depolymerization of HFn after treatment with 6M guanidine hydrochloride at 70 ° C for 10 min (mobile phase containing 6 M guanidine hydrochloride).
  • FIG. 64 Chromatographic peak position of HFn 100% depolymerization after treatment with 6M guanidine hydrochloride at 70 ° C for 30 min (mobile phase containing 6 M guanidine hydrochloride).
  • Figure 65 Chromatographic peak position of HFn 100% depolymerization after treatment with 6M guanidine hydrochloride at 70 ° C for 60 min (mobile phase containing 6 M guanidine hydrochloride).
  • FIG. 66 Chromatographic peak position of 67.8% depolymerization of HFn after treatment with 7M guanidine hydrochloride at 37 ° C for 60 min (mobile phase containing 7 M guanidine hydrochloride).
  • FIG. 67 Chromatographic peak position of 71.3% depolymerization of HFn after treatment with 7M guanidine hydrochloride at 37 ° C for 90 min (mobile phase containing 7 M guanidine hydrochloride).
  • FIG. 68 Chromatographic peak position of 75.4% depolymerization of HFn after treatment with 7M guanidine hydrochloride at 37 ° C for 120 min (mobile phase containing 7 M guanidine hydrochloride).
  • FIG. 69 Chromatographic peak position of 70.8% depolymerization of HFn after treatment with 7M guanidine hydrochloride at 37 ° C for 150 min (mobile phase containing 7 M guanidine hydrochloride).
  • Figure 70 Chromatographic peak position of 72.3% depolymerization of HFn after treatment with 7M guanidine hydrochloride at 37 ° C for 180 min (mobile phase containing 7 M guanidine hydrochloride).
  • FIG 71 Chromatographic peak position of HFn 100% depolymerization after treatment with 7M guanidine hydrochloride at 50 ° C for 60 min (mobile phase containing 7 M guanidine hydrochloride).
  • Figure 72 Chromatographic peak position of HFn 100% depolymerization after treatment with 7.5 M guanidine hydrochloride at 37 ° C for 60 min (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG. 73 Chromatographic peak position of HFn 100% depolymerization after treatment with 7.5 M guanidine hydrochloride at 37 ° C for 90 min (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG. 74 Chromatographic peak position of HFn 100% depolymerization after treatment with 7.5 M guanidine hydrochloride at 37 ° C for 120 min (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG. 75 Chromatographic peak position of HFn 100% depolymerization after treatment with 7.5 M guanidine hydrochloride at 37 ° C for 150 min (mobile phase containing 7.5 M guanidine hydrochloride).
  • FIG 77 The peak position of the HFn complex after the removal of guanidine hydrochloride (mobile phase 50 mM Tris).
  • the ferritin of the present invention refers to any ferritin which can form a cage structure, which may be a naturally-derived ferritin or a recombinantly expressed ferritin, or a mutant thereof, which may be derived from prokaryotes, protists, A fungus, plant or animal, for example derived from bacteria, fungi, insects, reptiles, birds, amphibians, fish, mammals, for example from rodents, ruminants, non-human primates or humans, eg small Rat, rat, guinea pig, dog, cat, cow, horse, sheep, monkey, gorilla, human.
  • the ferritin of the invention is human ferritin, and in some embodiments, the ferritin of the invention is a genetically engineered fully human heavy chain ferritin having the amino acid sequence set forth in SEQ ID NO. In other embodiments, the amino acid sequence of the ferritin of the invention is set forth in SEQ ID No. 2 or 3.
  • the denaturing agent of the present invention refers to a substance which depolymerizes ferritin, and is an anthraquinone hydrochloride or an anionic surfactant typified by SDS, or urea.
  • An anionic surfactant is an amphiphilic molecule containing both a hydrophilic group and a hydrophobic group, and can dissociate an anion in a solution state, including, for example, SDS, stearic acid, sodium dodecylbenzenesulfonate Wait.
  • the depolymerization of the present invention means that the tight closed spherical structure of the ferritin natural 24-mer is opened, and the visual representation is that the peak time on the gel exclusion column is significantly delayed, so that the depolymerized protein peak and the unagglomerated protein peak occur. Separation.
  • the depolymerization ratio is at least 50%, such as at least 55%, 60%, 65%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, up to achievable 100%.
  • Repolymerization means that after the substance causing depolymerization is removed or reduced to a sufficiently low concentration, the depolymerized ferritin returns to the tightly closed globular structure of the natural 24-mer, which is visually represented as a gel-exhaust column.
  • the peak time is reduced to a peak time similar to that of natural ferritin.
  • the polycondensation ratio is at least 50%, such as 55%, 60%, 65%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, up to 100%.
  • the drug or the carrier drug of the present invention means any drug which can be contained in ferritin as long as the molecular size of the drug is less than 8 nm.
  • the medicament or the encapsulating medicament of the present invention is selected from the group consisting of a tumor antibiotic, a natural source antineoplastic, a metal compound, a radioisotope, an alkylating agent, an antimetabolite antineoplastic, and a hormonal antitumor drug.
  • the antibiotic antineoplastic agent is selected from the group consisting of doxorubicin (doxorubicin hydrochloride), zorubicin hydrochloride, valrubicin, bleomycin sulfate, mitomycin, epirubicin hydrochloride, and edetate hydrochloride Bixing, actinomycin D, phoroxin, daunorubicin, pirarubicin, epirubicin, idarubicin, aclaramicin, bleomycin A5, chromomycin A3, hydrochloric acid Bleomycin, palmitomycin, pingyangmycin hydrochloride, daunorubicin, arubicin hydrochloride, nitrozamycin, piramycin, pirarubicin hydrochloride, actinomycin C; natural source
  • the antineoplastic agent is selected from the group consisting of topotecan hydrochloride, 10-hydroxycamptothecin, 7-ethyl-10-hydroxy
  • the drug-loaded ferritin of the present invention can be targeted to a tumor, and after binding to the tumor, the encapsulated drug is released, and the drug acts on the tumor to achieve prevention and/or treatment of the tumor.
  • human ferritin may specifically target human solid tumors and hematological malignant cells such as lung cancer, breast cancer, prostate cancer, cervical cancer, and colorectal by binding to its receptor transferrin receptor (Tferrol Receptor 1, TfR1). Cancer, ovarian cancer, esophageal cancer, gastric cancer, thymic cancer, T lymphocytic leukemia, erythrocyte leukemia, and the like.
  • the medicament or the entrapped drug of the invention is selected from a drug other than an antineoplastic drug, ie, a non-antitumor drug.
  • a drug may be a drug that does not require targeting, such as a systemically administered drug, such as a drug that is sparingly soluble, unstable, and/or susceptible to interaction and failure.
  • such a drug is selected from the group consisting of amphotericin B, glatiramer acetate, complex sodium gluconate, rapamycin, sevelamer sulfate binding agent, verteporfin for injection, sucrose Iron, peginterferon alfa-2a/2b, fenofibrate, pegetastatin, rifafil statin, amikacin, fentanyl, cyclosporine, cetirizine , capsaicin, ceramide, etc.
  • the non-antitumor drug of the invention is selected from the group consisting of a radiopharmaceutical, a neurotransmitter, a dopamine receptor agonist, a neurological central anticholinergic, a choline receptor agonist, a gamma secretase inhibitor, an antibiotic Oxygen or anesthetic
  • the radiopharmaceutical is selected from the group consisting of 64 Cu, 235 U
  • the neurotransmitter is selected from the group consisting of carbachol, atropine, scopolamine, dopamine and derivatives thereof
  • the dopamine receptor agonist is selected from the group consisting of bromocryption Ergolide derivatives such as kiosk, pergolide, and apomorphine, and non-ergoline derivatives
  • the central nervous anticholinergic agent is selected from the group consisting of trihexyphenidate, benzaltropine and propandidine
  • the choline receptor agonist is selected.
  • the ⁇ -secretase inhibitor is selected from the group consisting of difluoroketones
  • the antioxidant is selected from melatonin
  • the anesthetic is selected from the group consisting of guanamine.
  • an additive is added to the incubation solution when encapsulating the drug into ferritin to facilitate dissolution of the drug and/or aggregation of ferritin to achieve and/or improve drug loading.
  • a platinum-based drug is involved, in which case DMA, DMF, DMSO, or a mixture thereof, is required to be added to the incubation solution.
  • the additives that need to be added may also be different.
  • the mass ratio of ferritin to entrapped drug in the incubation solution is from 5:1 to 1:5, for example, 5:1 to 1:2, 2:1 to 1:4, 4:1 to 1: 1,8:2.5 to 8:3.5, 1:1 to 1:3, 1:1.5 to 1:2.5.
  • the mass ratio of ferritin to the drug may also vary.
  • the buffer solution of the incubation solution is Tris-HCl buffer, phosphate buffered saline (PBS), carbonate buffer, glycine buffer or citrate buffer, and the pH of the buffer is between 3 and 3 10, for example 4-9, 5-8.5, the concentration of the buffer is between 20 and 500 mM. Buffering capacity, pH range, desired concentration, etc. may need to be adjusted depending on the buffer used.
  • the upper limit of the drug-loaded drug in the ferritin of the present invention is the maximum amount of drug that can be contained in the internal volume of the ferritin cage structure, i.e., the amount of drug that can be contained in a volume of about 268 cubic nanometers.
  • the drug loading is 5 to 60%, preferably 10 to 50%, 20 to 40%, 25 to 35%, such as at least 8%, 10%, 15%, in terms of mass ratio. 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60% or higher.
  • 50 to 500 doxorubicin molecules or similarly sized molecules are entrapped per molecule of ferritin, eg, 60, 80, 90, 100, 110, 120, 130, 140, 150, 170, 190, 200 220, 240, 260, 280, 240, 260, 280, 300, 350, 400, 450, 500 or more.
  • ferritin eg, 60, 80, 90, 100, 110, 120, 130, 140, 150, 170, 190, 200 220, 240, 260, 280, 240, 260, 280, 300, 350, 400, 450, 500 or more.
  • the method for encapsulating a drug with ferritin of the invention can greatly increase the drug entrapment amount, and the clinical administration dose will be greatly reduced under the premise of achieving the same therapeutic effect.
  • the drug-loaded ferritin of the present invention may be provided in the form of a pharmaceutical composition, that is, in addition to the drug-loaded ferritin, which further contains a pharmaceutically acceptable carrier.
  • a pharmaceutical composition may be formulated according to conventional techniques using pharmaceutically acceptable carriers or diluents, as well as any other known excipients, such as those disclosed in Remington: The Science and Practice of Pharmacy, 22nd edition, edited by Gennaro, Mack Publishing Co., 2013.
  • the drug-loaded ferritin obtained by the method of the present invention can be used to treat and/or prevent a disease or condition in a subject.
  • the disease or condition in which the drug-loaded ferritin of the present invention can treat and/or prevent depends on the drug to be contained.
  • the disease or condition that the doxorubicin-coated HFn of the present invention can treat and/or prevent depends on the doxorubicin entrapped, ie, known in the art.
  • the disease or condition that can be treated and/or prevented by doxorubicin can be treated and/or prevented by the doxorubicin-encapsulated HFn of the invention.
  • the doxorubicin-encapsulated HFn of the present invention can be targeted to a tumor to release the entrained doxorubicin at the tumor site after administration to achieve prevention and/or treatment of the tumor.
  • the disease or condition is selected from the group consisting of acute leukemia (lymphocytic and granulocyte), malignant lymphoma, breast cancer, bronchial lung cancer (undifferentiated small cell and non-small cell), ovarian cancer, Soft tissue sarcoma, osteosarcoma, rhabdomyosarcoma, Ewing sarcoma, blastoma, neuroblastoma, bladder cancer, thyroid cancer, prostate cancer, head and neck squamous cell carcinoma, testicular cancer, stomach cancer, liver cancer.
  • acute leukemia lymphocytic and granulocyte
  • malignant lymphoma breast cancer
  • bronchial lung cancer undifferentiated small cell and non-small cell
  • ovarian cancer Soft tissue sarcoma, osteo
  • the disease or state in which the platinum-containing drug-containing HFn of the present invention can treat and/or prevent depends on the platinum-based drug to be contained, that is, for example, in the prior art. It is known that diseases or conditions that can be treated and/or prevented by DDP can be treated and/or prevented by the DDP-encapsulating HFn of the present invention. Without being bound by any theory, the platinum-loaded HFn of the present invention can be targeted to a tumor, thereby releasing the encapsulated platinum-based drug at the tumor site after administration to achieve prevention and/or treatment of the tumor.
  • the disease or condition is selected from the group consisting of small cell and non-small cell lung cancer, testicular cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, bladder cancer, melanoma, osteosarcoma, reticulum Cell sarcoma, seminoma, nasopharyngeal carcinoma, esophageal cancer, thyroid cancer, head and neck cancer, squamous cell carcinoma, and malignant lymphoma.
  • the disease or condition that can be treated and/or prevented by CBP can be treated and/or prevented by the HFn of the packaged CBP of the present invention, such as ovarian cancer, lung cancer, Head and neck cancer, germ cell tumor, thyroid cancer, cervical cancer, bladder cancer, etc.
  • compositions of this invention may be administered by any suitable route and mode, including, without limitation, intravenous or subcutaneous injection or infusion.
  • the pharmaceutical composition of the present invention may also be used in combination with other drugs, for example, other antitumor drugs different from the drug to be coated, such as vincristine, cyclophosphamide, 5-fluorouracil, or in combination with radiation therapy, for example ABVD, CAF, CAOP, FAM, AC, AOP, ACP, CY-VA-DIC, MACC.
  • other drugs for example, other antitumor drugs different from the drug to be coated, such as vincristine, cyclophosphamide, 5-fluorouracil, or in combination with radiation therapy, for example ABVD, CAF, CAOP, FAM, AC, AOP, ACP, CY-VA-DIC, MACC.
  • other drugs for example, other antitumor drugs different from the drug to be coated, such as vincristine, cyclophosphamide, 5-fluorouracil, or in combination with radiation therapy, for example ABVD, CAF, CAOP, FAM, AC, AOP, ACP
  • HFn was treated at different temperatures by using different concentrations of SDS (subunit sequence as shown in SEQ ID NO. 1) (specific conditions are shown in Table 1), and liquid phase (Agilent 1260 HPLC system, TSKgel G4000 gel exclusion efficiency is highly efficient) Liquid chromatography column) Observe whether this condition depolymerizes HFn.
  • the HFn was treated with 1% SDS, the liquid phase mobile phase was 1% SDS, the pH was 7.0, and the peak time was as shown in Table 1 and Figures 1-3. Because the SDS-PAGE gel electrophoresis sample was treated in a 2% SDS solution at 100 ° C for 10 min to depolymerize the protein sample, the depolymerization reference for this experiment was HFn treated in a 2% SDS solution at 100 ° C. 10min.
  • Peak time 1 Peak time 2 Depolymerization Depolymerization rate 2.0% 100 ° C 10min 18.838 Yes 100% 1.0% 30 ° C 40min 17.126 no 1.0% 30 ° C 80min 17.131 no 1.0% 50 ° C 30min 17.086 no 1.0% 50 ° C 60min 17.076 no 1.0% 70 ° C 15min 17.090 18.653 Yes 30% 1.0% 70 ° C 30min 17.093 18.675 Yes 30%
  • the peak time is 18.838min
  • the peak of 24.365min minutes through the ultraviolet 260/280 ratio to find the main component non-protein substances. Therefore, when the liquid phase mobile phase is 1% SDS, the peak of complete depolymerization is at 18.838min.
  • the HFn was treated with 2% SDS, the liquid phase mobile phase was 2% SDS, the pH was 7.0, and the peak time was as shown in Table 2 and Figure 4-6. Because the SDS-PAGE gel electrophoresis treatment of the sample is carried out in a 2% SDS solution at 100 ° C for 10 min, the protein sample can be depolymerized, so the depolymerization reference for this experiment is HFn in a 2% SDS solution at 100 ° C for 10 min. .
  • the peak time is 19.663min
  • the HFn was treated with 5% SDS, the liquid phase mobile phase was 5% SDS, the pH was 7.0, and the peak time was as shown in Table 3 and Figures 7-9. Because the SDS-PAGE gel electrophoresis treatment of the sample is carried out in a 2% SDS solution at 100 ° C for 10 min, the protein sample can be depolymerized, so the depolymerization reference for this experiment is HFn in a 2% SDS solution at 100 ° C for 10 min. .
  • Figure 9 can be seen 5% SDS, 30 ° C, 60 min; 5% SDS, 30 ° C, 120 min; 5% SDS, 50 ° C, 30 min; 5% SDS, 50 ° C, 60 min; 5% SDS HFn can be partially depolymerized by treatment of HFn at 5% SDS, 70 ° C, and 30 min.
  • the HFn depolymerization is about 60% at 30 ° C and about 80% at 70 ° C. It shows that the increase of temperature has a stronger effect on the depolymerization of HFn than time.
  • the HFn was treated with 7.5% SDS, the liquid phase mobile phase was 7.5% SDS, the pH was 7.0, and the peak time was as shown in Table 4 and Figures 10-12. Because the SDS-PAGE gel electrophoresis treatment of the sample is carried out in a 2% SDS solution at 100 ° C for 10 min, the protein sample can be depolymerized, so the depolymerization reference for this experiment is HFn in a 2% SDS solution at 100 ° C for 10 min. .
  • the HFn was treated with 10% SDS, the liquid phase mobile phase was 10% SDS, the pH was 7.0, and the peak time was as shown in Table 5 and Figures 13-15. Because the SDS-PAGE gel electrophoresis treatment of the sample is carried out in a 2% SDS solution at 100 ° C for 10 min, the protein sample can be depolymerized, so the depolymerization reference for this experiment is HFn in a 2% SDS solution at 100 ° C for 10 min. .
  • the optimal conditions for SDS to depolymerize HFn are 7.5% SDS, 70 ° C, and 15 min.
  • the main conditions affecting the depolymerization of HFn by SDS are temperature and SDS concentration.
  • the HFn was treated at 7.5% SDS, 70 ° C, 15 min, and then the treated sample was desalted using a G75 desalting column (or dialysis or ultrafiltration repolymerization), the desalted sample was collected, and the recombination was observed using a liquid phase.
  • the liquid phase mobile phase was 20 mM PB (20 mM Na 2 HPO 4 + 20 mM NaH 2 PO 4 ), pH 7.0.
  • HFn was formulated into a solution containing 1 mg/mL HFn, 7.5% SDS, pH 7.0, then water bath at 70 ° C for 15 min, then DOX was added to make the solution system contain 1 mg/mL HFn, 1 mg/mL DOX, 7.5% SDS, After incubating the solution at 4 ° C for 4 h at pH 7.0, the excess DOX and the SDS in the solution were removed using a G75 column, and the HFn sample was collected. (Buffer is 50mM Tris pH 7.2)
  • the molar ratio of DOX:HFn was 300:1, and the mass ratio was 30.20%.
  • the DOX:HFn molar ratio in the prior art was only 33:1, and the mass ratio was 3.7%.
  • HFn was treated with different concentrations of guanidine hydrochloride at room temperature (25 ° C) at a protein concentration of 1 mg/ml.
  • the percentage data in the table indicates the ratio of depolymerization.
  • the results show that the number of packages is 300 units of DOX per unit HFn.
  • the mass ratio is 30.20%, compared to the prior art, the DOX:HFn molar ratio is only 33:1, and the mass ratio is 3.7%.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Engineering & Computer Science (AREA)
  • Epidemiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • Nanotechnology (AREA)
  • Optics & Photonics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Physics & Mathematics (AREA)
  • Biomedical Technology (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Toxicology (AREA)
  • Inorganic Chemistry (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Medicinal Preparation (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne un procédé d'enrobage d'un médicament dans une ferritine et son produit associé. L'invention explore spécifiquement l'impact de la dépolymérisation et de la condensation en présence de ferritine, telle qu'une ferritine totalement humaine, lors de l'utilisation de différentes concentrations de SDS, d'hydrochlorure de guanidine ou d'urée impliquant différentes températures et/ou durées de traitement. Elle explore plus spécifiquement les conditions d'utilisation optimales de la ferritine pour l'enrobage d'un médicament, tel que la doxorubicine. Grâce aux conditions d'enrobage ainsi obtenues, l'enrobage du médicament dans la ferritine peut être rapide, convenable, efficace et suffisant.
PCT/CN2019/076895 2018-03-07 2019-03-04 Procédé d'enrobage d'un médicament dans une ferritine et produit associé WO2019170059A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201810188050.5 2018-03-07
CN201810188050.5A CN110237047B (zh) 2018-03-07 2018-03-07 铁蛋白包载药物的方法及其产物

Publications (1)

Publication Number Publication Date
WO2019170059A1 true WO2019170059A1 (fr) 2019-09-12

Family

ID=67846816

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2019/076895 WO2019170059A1 (fr) 2018-03-07 2019-03-04 Procédé d'enrobage d'un médicament dans une ferritine et produit associé

Country Status (2)

Country Link
CN (1) CN110237047B (fr)
WO (1) WO2019170059A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114259563A (zh) * 2021-12-27 2022-04-01 大连大学 一种硼递送剂及其制备方法
CN115998708A (zh) * 2022-07-04 2023-04-25 大连工业大学 一种可逆自组装蛋白包埋活性分子的方法

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111411120B (zh) * 2020-04-01 2022-04-08 中国农业大学 一种锌离子诱导铁蛋白自组装包埋活性小分子的方法
CN113332261B (zh) * 2021-05-18 2022-11-15 昆山新蕴达生物科技有限公司 铁蛋白包载药物及其制备方法
CN113476588B (zh) * 2021-07-06 2022-11-22 武汉大学 靶向乳腺癌的多功能纳米颗粒、其制备方法及其应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103212093A (zh) * 2012-01-19 2013-07-24 中国科学院地质与地球物理研究所 一种具有细胞靶向性的磁性纳米材料及其生物医学应用
CN104013599A (zh) * 2014-05-28 2014-09-03 中国科学院生物物理研究所 一种肿瘤特异性靶向给药的药物载体及其应用
CN104977404A (zh) * 2015-07-07 2015-10-14 宁波瑞源生物科技有限公司 抑制类风湿因子干扰的胶乳增强免疫比浊试剂
CN106110333A (zh) * 2016-07-11 2016-11-16 中国科学院过程工程研究所 一种以铁蛋白为载体的抗肿瘤药物及其制备方法

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7160855B2 (en) * 2002-03-14 2007-01-09 Children's Hospital & Research Center At Oakland Enhancement of iron chelation therapy
CN104667289B (zh) * 2014-01-28 2018-08-14 暨南大学 一种抗肿瘤药物载体及其使用方法

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103212093A (zh) * 2012-01-19 2013-07-24 中国科学院地质与地球物理研究所 一种具有细胞靶向性的磁性纳米材料及其生物医学应用
CN104013599A (zh) * 2014-05-28 2014-09-03 中国科学院生物物理研究所 一种肿瘤特异性靶向给药的药物载体及其应用
CN104977404A (zh) * 2015-07-07 2015-10-14 宁波瑞源生物科技有限公司 抑制类风湿因子干扰的胶乳增强免疫比浊试剂
CN106110333A (zh) * 2016-07-11 2016-11-16 中国科学院过程工程研究所 一种以铁蛋白为载体的抗肿瘤药物及其制备方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
LIANG, MINMIN ET AL.: "H-Ferritin-Nanocaged Doxorubicin Nanoparticles Specifically Target and Kill Tumors With a Single-dose Injection", PNAS, vol. 111, no. 41, 14 October 2014 (2014-10-14), pages 14900 - 14905, XP055202963, doi:10.1073/pnas.1407808111 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114259563A (zh) * 2021-12-27 2022-04-01 大连大学 一种硼递送剂及其制备方法
CN115998708A (zh) * 2022-07-04 2023-04-25 大连工业大学 一种可逆自组装蛋白包埋活性分子的方法

Also Published As

Publication number Publication date
CN110237047A (zh) 2019-09-17
CN110237047B (zh) 2021-12-21

Similar Documents

Publication Publication Date Title
WO2019170059A1 (fr) Procédé d'enrobage d'un médicament dans une ferritine et produit associé
Zhang et al. Convection enhanced delivery of cisplatin-loaded brain penetrating nanoparticles cures malignant glioma in rats
JP2750138B2 (ja) 細胞毒性薬との組合せにおける血管収縮性物質
JP2018522021A (ja) 治療用化合物の脳への一方向送達のためのインプラント組成物
JP5642892B2 (ja) 多形膠芽腫の治療のためのマシテンタンを含有する組み合わせ剤
KR20130092617A (ko) 암 치료에 사용하기 위한 베무라페닙 및 인터페론을 포함하는 병용 요법
EP1100475A1 (fr) Utilisation de nanoparticules chargees de medicament dans le traitement anticancereux
US8664268B2 (en) Gallium compositions for the treatment of liver cancer and methods of use
JPH11503460A (ja) ウィルスおよび癌の増殖を阻止するｎ−クロロフェニルカルバメートおよびｎ−クロロフェニルチオカルバメート含有医薬組成物
CN111658778B (zh) 药物组合物及其制备方法与应用
He et al. Toxicities and therapeutic effect of 5-fluorouracil controlled release implant on tumor-bearing rats
KR20180014042A (ko) 약물을 전달하기 위한 가교된 히알루론산 및 이를 이용한 약학적 제제
WO2022206451A1 (fr) Nanosupport de type à cage de ferritine à co-chargement ciblé de médicaments hydrophiles/hydrophobes et son utilisation
CN114699537B (zh) 一种改善缺氧增敏pd-1抗体疗效的ros响应抗癌药物
CN108135939A (zh) 用于治疗癌症的抗驱走性试剂和免疫治疗剂联合疗法和组合物
Thakur et al. Dual stimuli-responsive and sustained drug delivery NanoSensoGel formulation for prevention of cisplatin-induced ototoxicity
KR20220125213A (ko) (s)-이소프로필 2-((s)-2-아세트아미도-3-(1h-인돌-3-일)프로판아미도)-6-디아조-5-옥소헥사노에이트를 포함하는 정맥내 투여용 동결건조 조성물 및 이의 용도
CN101185629A (zh) 一种治疗实体肿瘤的地西他滨缓释剂
RU2535002C2 (ru) Средство коррекции отдаленных последствий нарушений сперматогенеза, вызванных цитостатическим воздействием
CN114569545B (zh) 一种稳定的米托蒽醌制剂
US20050266076A1 (en) Resorbable delivery systems for the treatment of cancer
CN110237262B (zh) HFn包载铂类药物的方法及其产物
CN108136021A (zh) 用于治疗癌症的抗驱走性试剂和抗癌剂联合疗法和组合物
Ferrari Un nuovo profarmaco a base di platino: il suo effetto antitumorale e nuovi approcci in vitro per comprendere nuovi target per il trattamento di tumori del sistema nervoso
US8835453B2 (en) Sustained release implant for granisetron

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: 19764293

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: 19764293

Country of ref document: EP

Kind code of ref document: A1