WO2015160271A1 - Composition lipidique utilisée pour la construction d'un support de médicament génétique liposomique ciblé par des anticorps, et son utilisation - Google Patents

Composition lipidique utilisée pour la construction d'un support de médicament génétique liposomique ciblé par des anticorps, et son utilisation Download PDF

Info

Publication number
WO2015160271A1
WO2015160271A1 PCT/PL2015/050011 PL2015050011W WO2015160271A1 WO 2015160271 A1 WO2015160271 A1 WO 2015160271A1 PL 2015050011 W PL2015050011 W PL 2015050011W WO 2015160271 A1 WO2015160271 A1 WO 2015160271A1
Authority
WO
WIPO (PCT)
Prior art keywords
mass
dspe
antibodies
peg
liposomes
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/PL2015/050011
Other languages
English (en)
Inventor
Monika TOPORKIEWICZ
Justyna MEISSNER
Kazimierz Kuliczkowski
Aleksander SIKORSKI
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Wroclawskie Centrum Badan EIT Sp zoo
Original Assignee
Wroclawskie Centrum Badan EIT Sp zoo
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 Wroclawskie Centrum Badan EIT Sp zoo filed Critical Wroclawskie Centrum Badan EIT Sp zoo
Publication of WO2015160271A1 publication Critical patent/WO2015160271A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/88—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation using microencapsulation, e.g. using amphiphile liposome vesicle
    • 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
    • A61K47/6905—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion
    • A61K47/6911—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion the form being a liposome
    • A61K47/6913—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion the form being a liposome the liposome being modified on its surface by an antibody
    • 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
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
    • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers
    • A61K9/1272—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers comprising non-phosphatidyl surfactants as bilayer-forming substances, e.g. cationic lipids or non-phosphatidyl liposomes coated or grafted with polymers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • 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
    • A61K47/186—Quaternary ammonium compounds, e.g. benzalkonium chloride or cetrimide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner

Definitions

  • Lipid composition used for construction of liposomal genetic drug carrier targeted with antibodies, and use thereof
  • the present invention relates to a lipid composition used for formation of genetic drug liposome carrier targeted with antibodies being used in particular in treatment of chronic lymphocytic leukemia.
  • Research was carried out at the University of Wroclaw, Faculty of Biotechnology, Department of Cytobiochemistry and at the Wroclaw Medical University in the Hematology Blood Neoplasms and Bone Marrow Transplantation Clinic.
  • Leukemia comprises a vast heterologous group of diseases of hematologic origin.
  • pluripotent bone marrow stem cells are subject to proliferation and differentiation into two main cell lines: myeloid and lymphoid.
  • myeloid cells mature into erythrocytes, monocytes, granulocytes, and blood platelets.
  • Lymphoid cells differentiate into B and T lymphocytes. Both the lymphoid and the myeloid line of stem cells can proliferate excessively on each stage of the differentiation process, resulting in an acute or chronic form of leukemia.
  • Acute leukemia is a rapidly progressing disease, wherein the altered cells lose their natural properties.
  • Chronic leukemia is characterized by slower development. Acute forms constitute ca. 50-60% of all leukemia cases, 20-30% are chronic lymphocytic leukemia forms, and 1 5-20% are chronic bone-marrow leukemia forms.
  • Chronic lymphocytic leukemia is one of the most common leukemia forms in adults in Europe and North America.
  • the most common leukemia type is the acute bone-marrow leukemia (AML, about 1 600 cases annually) and chronic lymphocytic leukemia CLL, appearing with a similar frequency, constituting about 1300 cases annually.
  • AML acute bone-marrow leukemia
  • CLL chronic lymphocytic leukemia
  • the standard method of treatment for these leukemia types is chemotherapy, however, about 20% of cases remain resistant to the effects of cytostatics.
  • Significant overexpression of BCL-2 gene has been found in leukemia cells, and the relation between the high level of BCL-2 gene expression and low effectiveness of chemotherapy was discovered.
  • a genetic drug is a DNA or RNA molecule, that is, principally, not able to penetrate the cell membrane barrier.
  • a significant problem is the sensitivity of "naked" nucleic acids molecules to nucleolytic enzymes present in the plasma and other tissue fluids. Due to its structure, a nucleic acid molecule also carries a high resultant negative charge promoting opsonization and removal from the bloodstream by reticuloendothelial system macrophages. Therefore, it is necessary to apply an appropriate carrier that will deliver the genetic drug into the target cell overcoming the above mentioned barriers.
  • a lipid composition for producing liposomes which contains natural and synthetic phospholipids, consisting of 38.7% by weight of egg yolk phosphatidylcholine, 1 3.5% by weight of dioleoylphosphatidylcholine, 5.4% by weight of DC-Cholesterol , 15.3% by weight of di-stearoylphosphoethanolamine derivative of polyethylene glycol (PEG2000), and 27% by weight of a trimethylammoniurn salt of 1 ,2-dioleoylpropane (DOTAP).
  • the composition due to the lack of an appropriate modification enabling ligand attachment, is not intended for selective delivery of genetic drug to the target cells.
  • a lipid composition which is intended to be used as an effective carrier of genetic drugs, containing EDOPC, DC-Cholesterol, DPhPE, and a liposome-nucleic acid complex.
  • the composition referred to demonstrates lower "capacity" of the genetic drug in relation to the present solution, being 2 to 20 ⁇ g of siRNA/ODN per 1 ⁇ of liposome casing lipids, compared to 1 1 to 32 ⁇ g of ODN per 1 ⁇ of lipids in the case of proposed below solution.
  • the mentioned above carrier is characterized by a very broad range of particle diameter values, 30-450 nm, where particularly such a large particles as > 100 nm are not desired during intravenous application of the carrier. Additionally, there are no experiments confirming effective genetic drug delivery by means of this carrier to target cells. The authors of the report tested only the effectiveness of a non-targeted variation of the carrier upon direct injection into the tumor..
  • the carrier must present protective properties towards the genetic drug encapsulated therein and at the same time, it must not change its therapeutic properties during long-term storage.
  • the application of the invention will enable selective and effective delivery of drugs to target cells, will reduce the expression level of selected genes, e.g. anti-apoptotic ones, and will sensitize the cells changed by the disease to the effects of low doses of cytotoxic drugs.
  • the object of the present invention is also obtaining a carrier of low nonspecific cytotoxic and hemolytic activity. Unexpectedly, the technical problems mentioned above have been solved by the present invention.
  • the first object of the present invention is a lipid composition used for construction of liposome carrier targeted with antibodies for genetic drug in the form of asODN, siRNA, miRNA, ribozyme RNA, or DNAzyme, characterized in that it contains from 35.2% to 45.7% by mass of phosphatidylcholine (PC), from 12.3 to 1 9.1 % by mass of dioleoylphosphatidylethanolamine (DOPE), from 4.9% to 6.4% by mass of 3p-[N-(Dimethylethane)carbamoyl]cholesterol (DC-CHOL), from 1 0.7% to 1 6.6% by mass of DSPE-PEG from 4.6% to 6.2% by mass of maleimide DSPE-PEG derivative (DSPE-PEG-Mal), and from 10.6% to 25.7% by mass of a trimethylammonium salt of 1 ,2-dioleoylpropane (DOTAP).
  • PC phosphatidylcholine
  • DOPE dioleoy
  • the composition according to the present invention is characterized in that it contains 42.5% by mass of PC, 14.8% by mass of DOPE, 5.9% by mass of DC-CHOL, 1 1 .95% by mass of DSPE-PEG , 5.05% by mass of DSPE-PEG-Mal, and 1 9.8% by mass of DOTAP.
  • the second object of the present invention is an use of the lipid composition specified in first object of the invention for production of a liposome carrier for genetic drugs for treating leukemia.
  • the liposome structure created this way provides protection to the genetic drug against degradation.
  • the genetic drug carrier enables effective and selective drug delivery to the disease changed cell, which makes it possible to minimize the side effects of the therapy. This condition can be satisfied by attaching the ligand targeting the carrier to the target cell.
  • the characteristic contents of maleimide groups of the modified DSPE-PEG-Mal lipid enables attachment of thiolated proteins and peptides, here - antibodies providing guiding the genetic drug to the target cells.
  • Effective attachment of antibodies to the surface of liposomes of the composition according to the present invention is subject to the presence of thiol groups in the antibody molecule (obtained in amine group thiolation with Traut's reagent) and the appropriate mole ratio of IgG to maleimide groups in the bi-layer of the carrier. What is also significant, is the fact that the obtained liposomes do not form aggregates in the suspension. The obtained liposome preparations do not show hemolytic activity towards human erythrocytes.
  • the targeted liposome carrier of the lipid composition according to the present invention demonstrates protective properties towards the genetic drug encapsulated therein.
  • fig. 1 represents a graph illustrating the liposome molecules diameter size distribution in the original preparation and after 12 months of storage in the form of suspension at the temperature of 4° C
  • fig. 2 represents a graph illustrating the liposomes zeta potential value distribution after 12 months of storage in the form of suspension at the temperature of +4°C
  • fig. 3 represents a diagram illustrating fluctuations in the nucleic acid content in liposomes during 12 months of storage in the form of suspension at the temperature of +4° C
  • fig. 4 represents a diagram illustrating the hemolytic properties of liposomes towards human erythrocytes, fig .
  • fig. 5 represents the results of electrophoretic analysis of the plasmid DNA encapsulated in the carrier
  • fig. 6 is a diagram illustrating the nonspecific cytotoxicity of the liposomes
  • fig. 7 represents a diagram illustrating the effectiveness of attaching antibodies to liposomes
  • fig. 8 represents graphs illustrating the selectivity and effectiveness of immuno-liposomes towards target and nontarget selected cell lines: L121 0, Jurkat T and Daudi
  • fig. 9 illustrates the penetration route and selectivity of immuno- liposomes
  • fig. 10 illustrates the effectiveness of BCL -2 gene expression silencing by immuno-liposomes in Raji (CD20+) leukemic cell line, fig.
  • fig. 12 represents a chart illustrating the effectiveness of treatment with immuno-liposomes containing asODN against BCL -2 gene in SCID mouse with tumors induced via subcutaneous implantation of Daudi CD20+ cells.
  • oligonucleotides (asODN, scODN, miRNA) were suspended in 1 50 ⁇ of deionized water, ddH20. This solution was supplemented with 300 ⁇ of chloroform . In order to obtain one phase, 300 ⁇ of methanol were added, and then it was titrated by drop, adding methanol until obtaining clear solution( ⁇ 100 ⁇ additionally). 150 ⁇ of DOTAP chloroform solution containing 2 mg of the lipid was added to the clear mixture. It was stirred gently to obtain a single phase. The mixture was left to rest for 30-40 minutes in room temperature.
  • the upper phase was removed, and the lower phase was added to the chloroform mixture of lipid solutions: 4.3 mg PC, 1 .5 mg DOPE, 0.6 mg DC-CHOL, 1 .1 9 mg DSPE-PEG , 0.51 mg DSPE-PEG-MAL, after which 250 ⁇ of PBS buffer was added (137 mM NaCI, 2,7 mM KCI, 10 mM Na 2 HP0 4 2 mM KH 2 P0 4 ). The mixture was subjected to sonication for 1 minute with concurrent cooling the sample in the ice bath.
  • the thiolation reaction of free amine groups was carried out using 1 .95 mg of antibodies (here: RituximabR) dissolved in 195 ⁇ of citrate buffer (C 6 H 5 0 7 Na 3 '2H 2 0, NaCI, NaOH, PolysorbateSO (Polyoxyethylene sorbitan monolaurate), and 0.036 mg of Traut's reagent in the presence of EDTA (20 mM) at pH 7.4, and then the whole reaction mixture was incubated for 4h in temperature of +4° C, stirring gently.
  • citrate buffer C 6 H 5 0 7 Na 3 '2H 2 0, NaCI, NaOH, PolysorbateSO (Polyoxyethylene sorbitan monolaurate)
  • Traut's reagent Polyoxyethylene sorbitan monolaurate
  • Unbound antibodies were separated from the immuno-liposomes by gel filtration on Sepharose 4B (column measuring 25 x 1 ,2 cm equilibrated with PBS buffer (137 mM NaCI, 2.7 mM KCI, 1 0 mM Na 2 HP0 4 , 2 mM KH 2 P0 4 ) at the temperature of +4° C.
  • PBS buffer 137 mM NaCI, 2.7 mM KCI, 1 0 mM Na 2 HP0 4 , 2 mM KH 2 P0 4
  • NOD/SCI D mice were subcutaneously implanted with Daudi (CD20+) line leukemic cells. Suspension of cells coming from in vitro culture was used for subcutaneous transplantation, in the amount of 1 *1 0 6 /mouse. After formation of subcutaneous tumor, the mice were treated with drug solutions in the form of liposome suspension according to the invention and water solution of mitoxantrone. Groups of 8 animals each were used for the experiment. Among the analyzed groups, 3 were control groups in order to exclude the therapeutic effects of separately acting low mitoxantrone dose of 0.1 mg/kg body mass, immuno-liposomes according to the invention, but containing the "scrambled" sequence, and PBS buffer.
  • the solutions of the drugs were prepared before use.
  • the preparations were administered in the volumes of 200-300 ⁇ (10 ⁇ /1 g body mass), wherein the dose each time contained, for immuno-liposome preparation : 800 Mg/kg body mass of ODN, 88 mg/kg body mass of lipids, 650 Mg/kg body mass Rituximab antibodies or 0.1 mg/kg body mass of mitoxantrone.
  • the solutions were injected into the lateral tail vein after immobilizing the animal.
  • the effectiveness of the treatment is presented in the chart illustrating the dependency of tumor growth from the time of treatment for each preparation administered as shown in fig.
  • lipid composition for production of liposome carrier for genetic drug in the form of asODN, siRNA or miRNA guarantees obtaining liposomes of stable size, with the diameter of about 1 10+/-5 nm, low growth dynamics during a year of storage in the form of suspension or freeze-dried, as illustrated by the chart presented in fig. 1 , size of which was measured by dynamic light scattering technique.
  • the obtained liposomes do not form aggregates in the suspension.
  • the obtained liposome preparation demonstrates stability in zeta potential of ca. 23 mV (fig.
  • the obtained liposome preparations do not show hemolytic activity towards human erythrocytes. Obtained hemolysis values of up to 7% remain within the error limits of the method (Fig. 4).
  • the hemolytic activity of the carrier was evaluated by examining the effects of hemoglobin from erythrocytes isolated from human blood after incubation with liposomes. Erythrocytes in the amount of 7*10 7 were incubated with the increasing amount of 0.5 - 4 Mg pDNA/ml encapsulated in the liposome carrier according to the present invention.
  • the blind sample consisted of erythrocytes suspended in PBS and incubated exactly asexamined samples. The total hemolysis was assumed to be 7*1 0 7 of cells suspended in deionized water and incubated as above. Hemoglobin leakage from liposome erythrocytes was the indicator of liposomes' hemolytic activity. Hemoglobin absorbance was measured at the wave length of 570 nm after 2 hours of incubation at 37 ° C.
  • the targeted liposome carrier of the lipid composition according to the present invention demonstrates protective properties towards the genetic drug encased therein.
  • Liposome carrier containing plasmid DNA was incubated with DNAse for 2 h at the temperature of +37° C, after which the protective properties of the liposomes were examined by means of electrophoretic analysis of the plasmid DNA encapsulated in the carrier (fig. 5).
  • liposome preparations In order to check the nonspecific cytotoxic activity of liposome preparations, the latter were tested against a broad spectrum of cell lines (Jurkat T lymphoblastic leukemia model, HL 60 myeloid leukemia, and HEL cells belonging to the erythroid line).
  • the liposome preparations of composition according to the present invention demonstrate low nonspecific cytotoxic activity (Fig. 6).
  • the liposomes cytotoxic activity was measured by means of MTT test (MTT Thiazolyl Blue Tetrazolium Bromide, Sigma Aldrich).
  • MTT test MTT Thiazolyl Blue Tetrazolium Bromide, Sigma Aldrich
  • the method uses the redox activity of mitochondria. In the effect of mitochondrial dehydrogenase, the yellow, water soluble tetrazolium salt is transformed into purple, insoluble in water crystals of formazan.
  • the amount of produced formazan crystals is proportional to the count of living cells.
  • the obtained immuno-liposome preparation (targeted with antibodies) demonstrates high selectivity towards target cells exposing surface CD20 antigen.
  • the selectivity of immuno-liposomes prepared according to the present invention was tested in vitro against L1210 (CD20), Jurkat T (CD20-), HeLa (CD20-), Daudi (CD20+), and RPMI 8226 (CD20+) cells, based on MTT toxicity tests (Fig. 8), and fluorescence imaging coming from liposomes marked with DiD (1 ,1 '-Dioctadecyl-3,3,3',3'- Tetramethylindodicarbocyanine, Invitrogen) (Fig.
  • the immuno-liposomes selectively reach only CD20+ cells, where the accumulation of immuno-liposomes fluorescently stained with DiD is visible in the area of the cell membrane of CD20+ cells (Fig. 9b).
  • the lipid composition of immuno-liposomes according to the present invention effectively reduces BCL-2 gene expression in Raji (CD20+) cells, which was evaluated on the basis of Bcl-2 protein level analysis in control cells and in those treated with immuno-liposomes, with Western Blot technique (Fig. 1 0).
  • the liposome preparation of the composition according to the present invention is also intended for application in genetic therapy combined with cytostatics, and therefore the effect of BCL-2 gene expression reduction upon survivability of Daud CD20+ cells and upon the effectiveness of combined therapy was also examined.
  • Reduction of BCL-2 gene expression by means of asODN in the immuno- liposomes according to the present invention significantly improves the effectiveness of the cytostatic, here: mitoxantrone, in the doses corresponding to IC 50 1 .25 nM (Fig. 1 1 ).
  • an appropriate lipid composition particularly increasing the content of DSPE-PEG, using maleimide-modified DSPE-PEG, selection of charges ratio between the cationic lipid and the genetic drug in the asODN form, and attachment of "targeting" antibodies, which in case of the tested system were humanized RituximabR (MabThera®) antibodies, enables selective and effective delivery of a genetic drug to leukemic cells containing CD20 surface marker.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Dispersion Chemistry (AREA)
  • Biochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Epidemiology (AREA)
  • Microbiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Hematology (AREA)
  • Immunology (AREA)
  • Oncology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Preparation (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention concerne une composition lipidique utilisée pour la création d'un support liposomique ciblé par des anticorps pour des médicaments génétiques sous forme d'ADN, de siARN, de miARN, d'ARN de ribozyme, et de DNAzyme, caractérisée en ce qu'elle contient de 35,2 % à 45,7 % en poids de phosphatidylcholine (PC), de 12,3 à 9,1 % en poids de dioléoylphosphatidyléthanolamine (DOPE), de 4,9 % à 6,4 % en poids de 3β-[N-(diméthyléthane)carbamoyl]cholestérol (DC-CHOL), de 10,7 % à 16,6 % en poids de DSPE-PEG, de 4,6 % à 6,2 % en masse de dérivé maléimide de DSPE-PEG (DSPE-PEG-Mal) et de 10,6 % à 25,7 % en masse d'un sel triméthylammonium de 1,2-dioléoylpropane (DOTAP), et son utilisation.
PCT/PL2015/050011 2014-04-18 2015-04-18 Composition lipidique utilisée pour la construction d'un support de médicament génétique liposomique ciblé par des anticorps, et son utilisation Ceased WO2015160271A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
PL407949A PL233741B1 (pl) 2014-04-18 2014-04-18 Kompozycja lipidowa sluzaca do wytworzenia kierowanego za pomoca przeciwcial liposomowego nosnika lekow genetycznych oraz jej zastosowanie
PLPL407949 2014-04-18

Publications (1)

Publication Number Publication Date
WO2015160271A1 true WO2015160271A1 (fr) 2015-10-22

Family

ID=54324351

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/PL2015/050011 Ceased WO2015160271A1 (fr) 2014-04-18 2015-04-18 Composition lipidique utilisée pour la construction d'un support de médicament génétique liposomique ciblé par des anticorps, et son utilisation

Country Status (2)

Country Link
PL (1) PL233741B1 (fr)
WO (1) WO2015160271A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020107088A1 (fr) * 2018-11-27 2020-06-04 Universidade Estadual De Campinas Procédé microfluidique d'obtention de liposomes stealth cationiques
CN111904934A (zh) * 2020-09-21 2020-11-10 中国医学科学院医药生物技术研究所 miRNA185抑制剂的脂质体及其制法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008120914A1 (fr) 2007-03-29 2008-10-09 Korea University Industrial & Academic Collaboration Foundation Composition de nanoparticules de phospholipides cationiques utilisée pour la livraison efficace d'acides nucléiques
WO2009031911A1 (fr) 2007-09-06 2009-03-12 Uniwersytet Wroclawski Vecteur de liposome cationique pour médicaments génériques
WO2009120247A2 (fr) 2007-12-27 2009-10-01 The Ohio State University Research Foundation Compositions de nanoparticules de lipides et procédés de préparation et d’utilisation de celles-ci
US20130202687A1 (en) * 2010-10-18 2013-08-08 Istituto Giannina Gaslini Lipid vectors delivering gene silencers

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008120914A1 (fr) 2007-03-29 2008-10-09 Korea University Industrial & Academic Collaboration Foundation Composition de nanoparticules de phospholipides cationiques utilisée pour la livraison efficace d'acides nucléiques
WO2009031911A1 (fr) 2007-09-06 2009-03-12 Uniwersytet Wroclawski Vecteur de liposome cationique pour médicaments génériques
WO2009120247A2 (fr) 2007-12-27 2009-10-01 The Ohio State University Research Foundation Compositions de nanoparticules de lipides et procédés de préparation et d’utilisation de celles-ci
US20130202687A1 (en) * 2010-10-18 2013-08-08 Istituto Giannina Gaslini Lipid vectors delivering gene silencers

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
DENG LI ET AL: "Comparison of anti-EGFR-Fab' conjugated immunoliposomes modified with two different conjugation linkers for siRNA delivery in SMMC-7721 cells", INTERNATIONAL JOURNAL OF NANOMEDICINE, vol. 8, 2013, pages 3271 - 3283, XP002743335 *
KIM K S ET AL: "Targeted gene therapy of LS174T human colon carcinoma by anti-TAG-72 immunoliposomes", CANCER GENE THERAPY, vol. 15, no. 5, May 2008 (2008-05-01), pages 331 - 340, XP002743336, ISSN: 0929-1903 *
ROTHDIENER M ET AL: "Targeted delivery of SiRNA to CD33-positive tumor cells with liposomal carrier systems", JOURNAL OF CONTROLLED RELEASE, vol. 144, no. 2, June 2010 (2010-06-01), ELSEVIER NLD, pages 251 - 258, XP002743334, DOI: 10.1016/J.JCONREL.2010.02.020 *
YU BO ET AL.: "Supplementary Material for Targeted nanoparticle delivery overcomes off-target immunostimulatory effects of oligonucleotides and improves therapeutic efficacy in chronic lymphocytic leukemia", BLOOD, January 2013 (2013-01-01), pages 1 - 21, XP002743506 *
YU BO ET AL: "Targeted nanoparticle delivery overcomes off-target immunostimulatory effects of oligonucleotides and improves therapeutic efficacy in chronic lymphocytic leukemia", BLOOD, vol. 121, no. 1, January 2013 (2013-01-01), pages 136 - 147, XP002743337 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020107088A1 (fr) * 2018-11-27 2020-06-04 Universidade Estadual De Campinas Procédé microfluidique d'obtention de liposomes stealth cationiques
CN111904934A (zh) * 2020-09-21 2020-11-10 中国医学科学院医药生物技术研究所 miRNA185抑制剂的脂质体及其制法
CN111904934B (zh) * 2020-09-21 2022-02-25 中国医学科学院医药生物技术研究所 miRNA185抑制剂的脂质体及其制法

Also Published As

Publication number Publication date
PL407949A1 (pl) 2015-10-26
PL233741B1 (pl) 2019-11-29

Similar Documents

Publication Publication Date Title
Du et al. Extracellular vesicles: a rising star for therapeutics and drug delivery
JP7600208B2 (ja) 拘束された脂質を含むナノ材料およびその使用
Yang et al. Functional exosome-mimic for delivery of siRNA to cancer: in vitro and in vivo evaluation
ES3031941T3 (en) Targeted lipid particles for systemic delivery of nucleic acid molecules to leukocytes
Xu et al. Osteoclast-targeted delivery of anti-miRNA oligonucleotides by red blood cell extracellular vesicles
CN114099533A (zh) 核酸药物递送系统、制备方法、药物组合物和应用
KR101085203B1 (ko) 의약 전달용 인지질 나노입자
Meissner et al. Novel antisense therapeutics delivery systems: In vitro and in vivo studies of liposomes targeted with anti-CD20 antibody
JP2017520549A (ja) 核酸の送達のためのリポソーム製剤
Safaei et al. Enzyme-sensitive nanoparticles, smart TAT and cetuximab conjugated immunoliposomes to overcome multidrug resistance in breast cancer cells
Shin et al. M1-polarized macrophage-derived cellular nanovesicle-coated lipid nanoparticles for enhanced cancer treatment through hybridization of gene therapy and cancer immunotherapy
US20220287969A1 (en) Multilamellar rna nanoparticles
Sun et al. Monitoring the in vivo siRNA release from lipid nanoparticles based on the fluorescence resonance energy transfer principle
Liu et al. Study on biomimetic nano tumor targeted delivery system for chemotherapy-laser immunotherapy
WO2015160271A1 (fr) Composition lipidique utilisée pour la construction d'un support de médicament génétique liposomique ciblé par des anticorps, et son utilisation
CN110898231A (zh) 一种功能化拉洛他赛脂质体及其制备方法与应用
WO2020232701A1 (fr) Système d'administration de médicament à nanoliposome marqué par un monosaccharide, sa méthode de préparation et son utilisation en tant que vecteur d'administration de ciblage pour un médicament
CN104922067A (zh) 一种载药纳米脂质体、制备方法及用途
WO2015160270A1 (fr) Forme liposomale ciblée de complexe d'oligonucléotide-polyéthylèneimine, son utilisation et son procédé d'obtention
EP4359542A1 (fr) Procédés et matériaux pour le traitement du cancer
US20120244210A1 (en) Composition for suppressing expression of target gene
US10639372B2 (en) Nucleic acid, medical nanoparticle, and pharmaceutical composition thereof
US20250339539A1 (en) Phosphate Membrane Nanodiscs Conjugated to Therapeutic Agents and Medical Uses Thereof
TWI734987B (zh) 單醣標記之奈米脂質體藥物遞送系統，其製法及其作為藥物靶定遞送載體之應用
Crow et al. The potential of small extracellular vesicles for pancreatic cancer therapy

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: 15724385

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: 15724385

Country of ref document: EP

Kind code of ref document: A1