US20050164963A1 - Components for producing amphoteric liposomes - Google Patents

Components for producing amphoteric liposomes Download PDF

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US20050164963A1
US20050164963A1 US10/505,093 US50509305A US2005164963A1 US 20050164963 A1 US20050164963 A1 US 20050164963A1 US 50509305 A US50509305 A US 50509305A US 2005164963 A1 US2005164963 A1 US 2005164963A1
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acid
liposomes
lipid
amphoteric
substance
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Frank Essler
Steffen Panzner
Gerold Endert
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Marina Biotech Inc
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Novosom AG
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Priority to US13/291,650 priority Critical patent/US8580297B2/en
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Priority to US14/052,215 priority patent/US9668973B2/en
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    • 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—Liposomes
    • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
    • 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
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    • A61K9/127—Liposomes
    • 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
    • A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
    • 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
    • A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
    • 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—Liposomes
    • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
    • A61K9/1272—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/06—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton
    • C07C229/10—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings
    • C07C229/16—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings to carbon atoms of hydrocarbon radicals substituted by amino or carboxyl groups, e.g. ethylenediamine-tetra-acetic acid, iminodiacetic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/24—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having more than one carboxyl group bound to the carbon skeleton, e.g. aspartic acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/64—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with substituted hydrocarbon radicals attached to ring carbon atoms, e.g. histidine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/645—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having two nitrogen atoms as the only ring hetero atoms
    • C07F9/6503—Five-membered rings
    • C07F9/6506—Five-membered rings having the nitrogen atoms in positions 1 and 3
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06191—Dipeptides containing heteroatoms different from O, S, or N
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0827—Tripeptides containing heteroatoms different from O, S, or N
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    • 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
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T428/00—Stock material or miscellaneous articles
    • Y10T428/29—Coated or structually defined flake, particle, cell, strand, strand portion, rod, filament, macroscopic fiber or mass thereof
    • Y10T428/2982—Particulate matter [e.g., sphere, flake, etc.]
    • Y10T428/2991—Coated

Definitions

  • the invention relates to amphoteric compounds based on amphiphilic molecules, the head groups of which being substituted with one or more amphoteric groups having an isoelectric point between 4 and 9, to liposomes containing such compounds, and to the use thereof.
  • lipids summarizes three classes of natural materials which can be isolated from biological membranes: phospholipids, sphingolipids, and cholesterol, including its derivatives. Industrially produced compounds with similar properties are diacylglycerols or N,N-dialkylamines.
  • liposomes can be used as containers for active substances in pharmaceutical preparations.
  • efficient and stable packaging of the cargo and controllable release of the contents are desirable. Both of these requirements are not easy to combine: the more stable and compact the packaging, the more difficult the release of the entrapped active substance therefrom.
  • liposomes changing their properties in response to an external stimulus have been developed.
  • Thermosensitive and pH-sensitive liposomes are well-known.
  • the pH-sensitive liposomes are of special interest, because this parameter undergoes changes even under physiological conditions, e.g. during endocytotic reception of a liposome in a cell, or during passage of the gastrointestinal tract.
  • pH-sensitive liposomes particularly comprise cholesterol hemisuccinate (CHEMS).
  • Cholesterol hemisuccinate in mixture with phosphatidyl ethanolamine, is used to produce pH-sensitive liposomes (Tachibana et al. (1998); BBRC 251, 538-544, U.S. Pat. No. 4,891,208).
  • Such liposomes can enter cells by endocytosis and are capable of transporting cargo molecules into the interior of cells on this route, without doing damage to the integrity of the cellular membrane.
  • CHEMS CHEMS anionic character. Liposomes produced using same have a negative overall charge, being taken up by cells with low efficiency. Despite the transfer mechanism described above, they are barely suitable for the transport of macromolecules into cells. Furthermore, packaging of macromolecular active substances such as DNA in such liposomes is not possible.
  • cationic liposomes having a preferably high and constant surface charge.
  • the positive overall charge of such particles leads to electrostatic adherence to cells and subsequently to efficient transport into same.
  • the use of these compounds and of liposomes produced using same remains restricted to in vitro or ex vivo applications, because such positively charged liposomes result in uncontrolled formation of aggregates with serum components.
  • WO 00/59474 the prior art describes compounds having a membrane anchor and a cationic and an anionic head group on the same molecule, the anionic group being linked to the basic structure via a disulfide bridge.
  • the disulfide bridge can be reduced under physiological conditions, e.g. by contact with the cytosol, the anionic head group then is liberated, and the overall molecule assumes a positive charge, thereby enabling fusion with the cell membrane.
  • the toxicity profile and storage stability of the compounds disclosed in WO 00/59474 are disadvantageous, because cleavage of the disulfide bridges results in free cationic lipids. Disadvantageously, these compounds are known to have a cytotoxic effect.
  • lipids and liposomes comprising same are disadvantageous in that the amounts of proteins, DNA and/or RNA which can be bound or entrapped by same are below average.
  • liposomes When modifying the liposomes so as to make them bind or entrap higher amounts of cargo, they will become cytotoxic or have low compatibility with serum or blood, or will be unstable in blood or serum because they would be attacked by particular components of the blood or serum, such as complement or perforin.
  • Well-known liposomes and lipids therefore have only limited suitability for use in living organisms and, in addition, exhibit low efficiency in conveying ingredients and/or binding substances.
  • the invention solves the above technical problem by means of liposomes comprising amphiphilic compounds according to general formula (I), which compounds have an isoelectric point between 4.5 and 8.5 and are capable of reversibly changing their state of charge from cationic to anionic within a pH range of 1-2 units: Amphoteric substance-Y-spacer-amphiphilic substance (I) wherein
  • the invention therefore relates to the teaching that conjugation of amphoteric groups via a spacer to an amphiphilic substance which can be incorporated in liposomal membranes allows providing structures which, in particular, can be used in the production of liposomes suitable for use in a living organism and capable of binding and/or conveying large amounts of proteins, peptides, carbohydrates, DNA and/or RNA.
  • the new compounds are suitable in the production of vesicles or liposomes which do not aggregate in blood or serum, are not attacked by complement components, are not cytotoxic, and are stable in blood or serum for several hours, the permeability of the liposomes depending on the pH value and thus on the state of charge of the compounds. That is, the release of active substances by these non-aggregating, stable, non-cytotoxic liposomes depends on the pH value of the medium.
  • Said one or two long-chain alkyls or acyls present in this molecular component comprise between 8 and 30 C atoms. They are preferably linear or slightly branched and may have 0, 1, or 2 ethylenically unsaturated bonds. Particularly preferred are substituents as found in natural lipids, i.e., straight-chain fatty acids or alcohols having 12 to 20 C atoms, with zero, one or two unsaturated bonds. Still more preferred are lauroyl, myristoyl, palmitoyl, stearoyl, oleoyl, and linoleoyl residues or the corresponding fatty alcohols thereof.
  • Diacylglycerols, dialkylglycerols, phosphoglycerols, acylated or alkylated 3-amino-1,2-propanediols, as well as N,N-dialkylamines are preferably employed as amphiphilic substances, because these compounds, in particular, are available at low cost, involve ordinary chemistry, and allow incorporation in membranes in high amounts without increasing the permeability thereof or even completely destroying their membrane character.
  • dicarboxylic acids are used as polar head group of the amphiphilic substance, conveniently allowing coupling of the actually charge-bearing substituents via additional functional groups.
  • amphiphilic substances derived therefrom are preferably long-chain esters of 1,4- or 1,5-dicarboxylic acids such as aspartic acid, glutamic acid, malic acid, tartaric acid, citric acid, aconitic acid, citraconic acid, maleic acid or similar compounds with fatty alcohols.
  • Particularly preferred are lauryl, myristyl, palmityl, stearyl, oleyl, and linoleyl esters of the above-mentioned dicarboxylic acids.
  • Additional molecular components spacer, amphoteric substance) are coupled via the remaining amino group, hydroxyl group, carboxylic group, or via the double bond.
  • diamines with an additional functional group e.g. in the form of a diamide of 3-aminoalanine, diaminobutyric acid, ornithine, or lysine with long-chain fatty acids.
  • additional functional group e.g. in the form of a diamide of 3-aminoalanine, diaminobutyric acid, ornithine, or lysine with long-chain fatty acids.
  • Particularly preferred amongst these are lauroyl, myristoyl, palmitoyl, stearoyl, oleoyl, and linoleoyl residues.
  • the compounds according to the invention can also be produced in the form of derivatives of sphingosine or ceramide. They can also be prepared as derivatives of long-chain vinyl ethers or of plasmalogens.
  • amphiphilic substances used with advantage as starting materials can be functionalized in their hydrophilic head group in various ways so as to conveniently allow stable, yet biologically degradable coupling or optionally assume the function of a spacer.
  • Particularly suitable for direct coupling is the hydroxyl group that is present, or an amino group.
  • Also suitable are carboxylic groups.
  • the overall molecule assumes its pH-dependent charge characteristics by the simultaneous presence of cationic and anionic groups in the “amphoteric substance” molecule portion. More specifically, an amphoteric substance is characterized by the fact that the sum of its charge components will be precisely zero at a particular pH value. This point is referred to as isoelectric point (IP). Above the IP the compound has a negative charge, and below the IP it is to be regarded as a positive cation, the IP of the compounds according to the invention ranging between 4.5 and 8.5.
  • a compound according to the invention is formed by coupling the amino group of histidine to dipalmitoylglycerol hemisuccinate.
  • the product has a negative charge because the carboxyl function which is present therein is in its fully dissociated form, and the imidazole function only has low charge.
  • an acid pH value of about 4 the situation is reversed: the carboxyl function now is largely discharged, while the imidazole group is essentially fully protonated, and the overall charge of the molecule therefore is positive.
  • the phosphate constant negative charge present therein has to be compensated by an additional positive charge in order to form a compound according to the invention.
  • One compound intended to illustrate the teaching of the invention is formed by coupling histidine to phosphatidyl serine. In this case, it is irrelevant whether coupling takes place between the carboxyl group of histidine and the amino group of PS or between the amino group of histidine and the carboxylic function of PS. In both cases, molecules are formed wherein a free amino group neutralizes the constant negative charge of the phosphate group. The remaining carboxylic group and the imidazole function react as above, producing the intended characteristics of the structures according to the invention.
  • the molecule has an isoelectric point between 4 and 8, preferably between 5 and 7.
  • the amphoteric substance has one or more cations with a pK a value of between 4 and 8 and, at the same time, one or more anions with a pK a value of between 3 and 7.
  • the amphoteric substances can be composed of two charge carriers which both alter their charge in the above-mentioned pH range of between 4 and 9. The simultaneously occurring loss of anionic charge and gain of cationic charge results in a change of charge in the overall molecule.
  • a particularly preferred embodiment comprises amphoteric substances where the pK a values of cation and anion are 2 pH units apart at maximum. This is particularly advantageous to the sharpness of the IP. The closer the two pK a are together, the more narrow the pH range wherein the molecule charge changes from cationic to anionic.
  • An advantageous selection as to type and number of cations and anions can be made by a person skilled in the art with reference to the above-mentioned formula.
  • functional groups or molecule fragments as charge carriers, which are in dissociated form in a pH range between 4 and 9. More specifically, these include phosphoric acid groups, sulfonic acid groups or other strong anions. Also preferred are most of the primary, secondary or tertiary amino groups, including quaternary ammonium, amidinium, pyridinium, and guanidino groups.
  • a particularly advantageous example of the above molecular component is the phosphoric acid group of phospholipids.
  • these fixed charges of the amphoteric substance must be overcompensated by the variable charges described above. This is only possible when using an excess of variable charge carriers.
  • a tertiary amine as cation for example, at least 2 carboxyl groups are required in order to obtain an amphoteric substance in the meaning of the invention. In case of only one carboxylic group, it is only possible to compensate the positive charge of the amine, and the molecule can never be completely recharged.
  • One advantage when using fully dissociated groups is their strong polarity.
  • amphoteric substances can be in the form of complete structural moieties.
  • this is the case with o-, m- or p-aminobenzoic acids, imidazolecarboxylic acid, imidazolediacetic acid, but also nicotinic acid or picolinic acid.
  • ⁇ -(1-piper-azino)alkylcarboxylic acids urocanic acid, 4-(2-amino-ethyl)imidazole-maleic acid monoamide, 4-(2-hydroxyethyl)-imidazole-maleic acid monoester, (2-aminoethyl)morpholine-maleic acid monoamide or analogous compounds.
  • the cation preferably is an imidazole, a piperazine, morpholine, purine, or pyrimidine.
  • Other advantageous cations having this property essentially include nitrogen bases. Particularly in those cases where the nitrogen bases are in the form of a ring system, positional isomers advantageously are existing, wherein the linking spacer is substituted on various positions of the organic cation.
  • the pK a values of the organic cations can be influenced via said positional isomerism.
  • the relevant fundamental rules are well-known to those skilled in the art. Alternatively, these effects can be estimated from tabular compilations (Handbook of Chemistry and Physics, Vol. 73, pp. 8-37ff.).
  • advantageous organic cations are represented by the following classes of substances:
  • the above-mentioned structural fragments may also have additional substituents.
  • these can be methyl, ethyl, propyl, or isopropyl residues, more preferably in hydroxylated form, including one or two hydroxyl groups. Also, these can be hydroxyl or keto functions in the ring system.
  • nitrogen bases with preferred pK a values are also formed by single or multiple substitution of an amine with lower alkanehydroxyls such as hydroxymethyl or hydroxyethyl groups.
  • Suitable organic bases from this group are e.g. aminopropanediols, triethanolamines, tris(hydroxy-methyl)methylamines, bis(hydroxymethyl)methylamines, tris-(hydroxyethyl)methylamines, bis(hydroxyethyl)methylamines, or the appropriately substituted ethylamines.
  • Nitrogen bases with preferred pK a values can also be found amongst aminosugars or aminosugar alcohols.
  • the anionic charge carriers are carboxylic groups.
  • any carboxylic acid can be used as charge carrier.
  • these include aliphatic straight-chain or branched carboxylic acids with up to 8 C atoms and 0, 1 or 2 ethylenically unsaturated bonds.
  • Exemplary components of compounds are the carboxyl group itself, acetic acid, bromoacetic acid, chloroacetic acid, acetoacetic acid, propionic acid, acrylic acid, butyric acid, crotonic acid, or higher carboxylic acids bound in the aliphatic chain, dicarboxylic acids such as oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, glutaric acid, adipic acid, caprylic acid, pimelic acid, suberic acid, cyclohexanedicarboxylic acid or cyclopentanedicarboxylic acid, mono-esterified or amidated or bound in the aliphatic chain, oligocarboxylic acids such as citric acid, isocitric acid or ethylenediaminetetraacetic acid, mono-esterified or amidated or bound in the aliphatic chain.
  • dicarboxylic acids such as oxalic acid, malonic
  • compositions are glycolic acid, lactic acid, hydroxybutyric acid, malic acid, tartaric acid, aspartic acid or glutamic acid, alanine, glycine, serine, threonine, asparagine, glutamine, proline, tyrosine or cysteine, or other amino acids or hydroxy acids, bound in the side chain via a heteroatom.
  • Carboxylic acids with suitable properties can also be found as substituents of aromatic systems, e.g. as benzoic acid, anisic acid, o-, m- or p-hydroxybenzoic acid, as dihydroxybenzoic acid, gallic acid, cinnamic acid, phenylacetic acid, hippuric acid, phthalic acid, terephthalic acid, 2-, 3- or 4-pyridinecarboxylic acid, furancarboxylic acid.
  • Other anionic groups are dissociable hydroxyls or thiols, such as occurring in ascorbic acid, N-substituted alloxan, N-substituted barbituric acid, in veronal, phenol, or as a thiol group.
  • the amphoteric substances are peptides including from 2 to 6 amino acids.
  • particularly the amino acids histidine, arginine, lysine, glutamic acid, or aspartic acid are used in a particularly preferred fashion to form the amphoteric substance and determine the charge characteristics thereof.
  • Other preferred amino acids are glycine, serine, threonine, glutamine, asparagine, but also cysteine, which contribute to increase the polarity and thus enhance the solubility of the amphoteric substance.
  • compositions of such peptides are shown in the following Table 1 as percentage of total amino acids: TABLE 1 Amino acid His Arg/Lys Asp/Glu Conditions i Up to 66% — Up to 66% His, Asp/Glu ⁇ 0 ii — ⁇ 50% >Arg/Lys Arg/Lys, Asp/Glu ⁇ 0 iii ⁇ 33% ⁇ 33% >Arg/Lys all ⁇ 0
  • i) illustrates the case of two pH-sensitive components, ii) the case of one fixed and one pH-sensitive component, and iii) the case of a mixture of i) and ii).
  • the sequence of the individual amino acids is arbitrary, the overall composition mainly determining the charge characteristics. Terminal group of the peptide are blocked in the form of an amide in the case of the C terminus, and with acetyl in the case of the N terminus.
  • the spacer is a lower alkyl residue of linear, branched or cyclic structure, which has from 0 to 8 C atoms and includes 0, 1 or 2 ethylenically unsaturated bonds.
  • the spacer may have hydroxyl groups so as to increase the polarity of the molecule.
  • the spacer can be a sugar, and advantageously a polyethylene glycol which may comprise up to 20 monomer units.
  • the linking group X comprises the structures —(C ⁇ O)—O—; —(C ⁇ O)—NH—; —NH—(C ⁇ O)—O—; —O—; —NH—; —CH ⁇ N— or —S—S—.
  • the linking group Y may correspond in its structure to the group X, and may additionally comprise the structures —O—(O ⁇ C)—; —S—(O ⁇ C)—; —NH—(O ⁇ C)—; —O—(O ⁇ C)—NH—; or —N ⁇ CH—.
  • X and/or Y can also be deletions, i.e., their presence is not required.
  • the Y group can be omitted in those cases where the amphoteric substance can be coupled directly to the amphiphilic substance, e.g. in the esterification of imidazole-4,5-dicarboxylic acid with dipalmitoylglycerol.
  • Particularly preferred molecules can be prepared by
  • Particularly preferred compounds include the following embodiments (long-chain hydrocarbon chains of the amphiphilic substances are represented in abbreviated spelling, corresponding to lauroyl, myristoyl, palmitoyl, stearoyl, oleoyl and linoleoyl residues):
  • N-Phosphatidyl ethyl-3-amino-5- imidazoylcarboxylic acid a) addition of phosphatidyl ethanolamine to urocanic acid according to iii) or b) according to vi) with ⁇ -histidine and phosphatidyl glycerol #31 Acetyl-Lys-Ala-His coupled via the side chain amino group of Lys to DPPG according to vi)
  • the long-chain amphiphilic substances are represented in abbreviated spelling, corresponding to lauryl, myristyl, palmityl, stearyl, oleyl, and linoleyl residues.
  • #50 N-(Aspartic acid dihydroxyalkyl)- 3-amino-5-imidazolecarboxylic acid, can be prepared by addition of aspartic acid diester to urocanic acid.
  • amphoteric components include e.g. the following compounds wherein R 1 and R 2 represent the amphiphilic substance and ( )n additional molecule portions according to the spacer defined above.
  • Histidine derivatives Coupling of the amphiphilic substance preferably proceeds via the amino group as R 2 .
  • R 1 is an anion and can be e.g. H or a hydroxycarboxylic acid or one or more amino acids.
  • R 2 is an anionic residue, e.g. a carboxylic acid or dicarboxylic acid.
  • Piperazine derivatives Coupling of the amphiphilic substance may proceed via one of the ring atoms.
  • Piperidine derivatives Coupling of the amphiphilic substance may proceed via any of the ring atoms. In those cases where the side chains are hydroxycarboxylic acids or amino acids, coupling may proceed with advantage via their heteroatoms.
  • Diaminobenzoic acid derivatives Coupling of the amphiphilic substance preferably proceeds via any of the two amino groups.
  • the second amino group can be alkylated, for example, so as to obtain a higher pK a value.
  • Coupling as an amide of phosphatidyl serine is another preferred embodiment of the invention.
  • Nitrilotriacetic acid derivatives Amphoteric groups are also formed by esterification of nitrilotriacetic acid.
  • N-Alkylcarboxyamino acid derivatives Amphoteric compounds are also formed by coupling of sterols to the terminal groups of N-acylamino acids.
  • the structure can be derived from serine, aspartic acid or glutamic acid, or from lysine or ornithine. Coupling of the aminodicarboxylic acids not only can be at the terminus, but also at the other acid groups.
  • the charge properties of such compounds can be # modified by complexing of metal ions.
  • EDTA derivatives Amphoteric groups are also formed by esterification of ethylene- diaminetetraacetic acid. In addition, the charge properties of complexing of metal ions.
  • the invention also relates to liposomes comprising the compounds according to the invention.
  • the compounds of the invention can be incorporated in high amounts in liposomal membranes to form amphoteric liposomes which are characterized in that their state of charge undergoes a reversible change when changing the pH of the surrounding medium.
  • the liposomes are cationic below their isoelectric point, and anionic above said point.
  • Liposomes comprising the components of the invention can be coated with polymers under suitable conditions, where single or multiple deposition of such substances on the surface is possible. In multiple deposition, optionally in the presence of crosslinkers, liposomal nanocapsules are formed. Methods of producing said liposomes are known to those skilled in the art e.g. from WO 00/28972 or WO 01/64330 which hereby are incorporated by reference and thus deemed part of the disclosure.
  • One particularly advantageous fact when using the substances according to the invention is that the electrostatic interaction with the polymer can be interrupted if said polymer is a polyelectrolyte.
  • the interaction of a polyelectrolyte with charge carriers of the liposomal membrane may give rise to demixing of membrane components and formation of lipid clusters. In many cases, such demixing is accompanied by a permeabilization of the liposomes.
  • the substances of the invention allow elimination of this interaction subsequent to the coating process.
  • the liposomes When increasing the pH value to or above the IP at this point in time, the liposomes will be entrapped in the nanocapsules merely in a steric fashion, and interaction between the membrane and the polyelectrolytes does no longer exist.
  • cluster formation of lipids and associated permeabilization of the membrane can be circumvented in this way.
  • liposomes including the substances of the invention in the membrane thereof readily undergo fusion with other membranes, particularly cell membranes, below the isoelectric point of the substance.
  • this step requires the presence of a major amount of PE in the membrane.
  • said PE assumes the function of a helper lipid.
  • the inferior stability of such membranes is disadvantageous, and gradual release of entrapped active substances is frequently observed.
  • liposomes produced using the substances according to the invention undergo effective fusion even in the absence of helper lipids.
  • the proportion of amphoteric lipids comprises 30 mole-%, 40 mole-% or 50 mole-% of total lipids at maximum.
  • Compositions comprising at least 2 mole-% of amphoteric lipids, but 50 mole-% at maximum, are particularly advantageous.
  • Compositions comprising at least 5 mole-% of amphoteric lipids, preferably 10 mole-%, and 50 mole-% at maximum, and preferably 40 mole-% are particularly preferred.
  • the production of liposomes comprising the substances of the invention proceeds according to techniques well-known to those skilled in the art.
  • the liposomes comprise phosphatidyl choline, phosphatidyl ethanolamine, diacylglycerols, ceramides, sphingolipids, tetraether lipids and/or PEG lipids.
  • the inventive compounds do not always form liposomes by themselves, and it may therefore be advantageous to add the above-mentioned lipids.
  • the liposomes have an average size between 50 and 1000 nm, preferably between 50 and 500 nm, more preferably between 50 and 300 nm and especially preferably between 60 and 130 nm.
  • liposomes which substances can be used e.g. in cancer therapy and in the therapy of severe infections.
  • liposome dispersions can be injected, infused or implanted. Thereafter, they are distributed in the blood or lymph or release their active substance in a controlled fashion as a depot. The latter can be achieved by highly concentrated dispersions in the form of gels.
  • the liposomes can also be used for topical application on the skin. In particular, they may contribute to improved penetration of various active substances into the skin or even passage through the skin and into the body.
  • the liposomes can also be used in gene transfer. Due to its size and charge, genetic material is usually incapable of entering cells without an aid.
  • suitable carriers such as liposomes or lipid complexes are required which, together with the DNA, are to be taken up by the respective cells in an efficient and well-directed fashion.
  • the active substance is a protein, a peptide, a DNA, RNA, an antisense nucleotide, and/or a decoy nucleotide.
  • liposomes are highly similar to cell membranes. Therefore, they can be used as membrane models to quantify the permeation rate of active substances through membranes or the membrane binding of active substances.
  • At least 50 ⁇ g, more advantageously 100 ⁇ g, preferably 150 ⁇ g of active substance per mg lipid is entrapped in the liposomes.
  • non-incorporated cargo molecules adhering on the outside can be removed by simply increasing the pH value. This step is necessary in all those cases where non-incorporated cargo molecules would give rise to aggregation of the liposomes.
  • One advantageous fact when using the components of the invention is that the entrapped active substances must be maintained under conditions allowing interaction with the lipid layer only during the period of actual enclosure. Once the lipid layer remains closed in itself, it is possible to change to other conditions. Thereby, possible inactivation of active substances, particularly of proteins, can be minimized.
  • the invention also relates to methods of loading liposomes with active substances, using a binding pH value for encapsulation and a second pH value to remove unbound active substances.
  • the invention relates to a method of loading liposomes with active substances, wherein the liposomes are made permeable at a specific pH value and subsequently sealed.
  • changes in permeability preferably can be used in a well-directed fashion in loading liposomes.
  • an active substance to be enclosed can be added to a medium under conditions of high permeability, followed by adjusting conditions of low permeability. In this way, the active substance will remain inside the liposomes. Thereafter, non-entrapped active substance can be removed, if necessary.
  • Such changes in permeability can be induced on liposomes or on liposomal nanocapsules.
  • the invention also relates to the use of the liposomes in diagnostics and in release systems.
  • the liposomes can also be used in a detection system.
  • the liposomes can be loaded with metal ions whose fluorescence is enhanced by chelate formation, i.e., terbium or europium ions, for example.
  • Liposomes for such uses may of course include components determining the specificity, i.e., antibodies, lectins, selectins, receptors, or hormones, or RNA aptamers.
  • the presence of these metal ions is restricted to the volume of the liposomes so as to avoid non-specific signals from slowly released metal ions adhering on the outside. It is also convenient to use the liposomes in the production of nanocapsules.
  • the liposomes can be used with advantage in the production of release systems in diagnostics.
  • the liposomes can be used as depot formulation and/or as circulative depot.
  • the use of the liposomes as a vector to transfect cells in vivo, in vitro and/or ex vivo is also advantageous.
  • the liposomes can be used in intravenous and/or peritoneal application.
  • the compounds and liposomes according to the invention involve several advantages. Surprisingly, it has been determined that the permeability of the inventive liposomes depends on the pH value and thus, on the state of charge of said compounds.
  • Liposomes produced using the structures according to the invention are therefore particularly suited to construct release systems wherein release of active substances is to proceed in dependence on the pH value of the medium.
  • liposomes including e.g. histidinyl-PS or histidinyldiacylglycerol hemisuccinate in the membranes thereof are capable of chelating metal ions.
  • This property results in an increase of the positive charge of the liposome.
  • This effect is observed to be particularly strong at neutral pH values, because the inherent charge of the compound is low in this case. Owing to their chelating properties, such liposomes can be used in biochemical diagnostics and in pharmaceutical therapy.
  • One essential precondition for the use of liposomes for experimental or therapeutic purposes is their compatibility with cells and tissues.
  • a number of well-known compounds used to incorporate DNA or proteins in cells are cytotoxic.
  • some of the compounds of the invention exhibit reduced cytotoxicity.
  • liposomes to be used in protein or gene transfer are their stability under physiological conditions.
  • liposomes Upon application into the blood circulation, liposomes are attacked by components of the complement system and undergo rapid lysis. This reaction proceeds within minutes. As a result, pores are formed in the membrane, which allow even large molecules such as proteins to diffuse out, therethrough.
  • stabilization of liposomes with respect to this mechanism is only possible by incorporating cholesterol in the lipid layer. While such liposomes are highly stable, they are no longer able to interact with cells or readily release their active substance.
  • liposomes constructed using the components of the invention can be stable in serum or blood for several hours. Even under such conditions, the release of active substance is low.
  • a liposomal vector for the transport of active substances must satisfy at least three preconditions: it must have low toxicity, entrap the active substance firmly and stably, and be compatible with serum or blood.
  • liposomes produced using selected substances according to the invention are therefore well suited for therapeutic uses.
  • Other properties supporting such uses are good loadability with active substances and well-directed removal of these substances by changing the pH value or by permeabilization of the membrane.
  • Liposomes produced using the substances of the invention show low non-specific binding to cell surfaces. It is this low non-specific binding which is an essential precondition for achieving specific binding to target cells.
  • Target control of the vehicles is obtained when providing the above-described liposomes with additional ligands. As a result, the active substance can be accumulated specifically in such cells or tissues which exhibit a pathological condition.
  • the substances according to the invention are therefore in the construction of vectors for transfer of active substances in living organisms.
  • the vectors are particularly suited for the transport of therapeutic macromolecules such as proteins or DNA which themselves are incapable of penetrating the cell membrane or undergo rapid degradation in the bloodstream.
  • amphoteric amphiphilic substances with only one hydrocarbon chain such as disclosed in U.S. Pat. No. 6,255,344, are also suitable in the production of amphoteric liposomes according to the invention, provided the hydrocarbon chain has more than 12 CH 2 groups.
  • the above liposomes can be loaded with active substance, especially DNA or oligonucleotides, according to the above-mentioned process, and their use in the transfection of cells and in gene therapy is hereby incorporated in the disclosure of the present invention.
  • double-chain amphiphilic substances formed by amidation of long-chain fatty acids with long-chain ⁇ -aminocarboxylic acids and provided with an amphoteric group such as histidine are also suitable in the production of amphoteric liposomes.
  • liposomes can be loaded with active substance, especially DNA or oligonucleotides, according to the above-mentioned process, and their use in the transfection of cells and in gene therapy is hereby incorporated in the disclosure of the present invention.
  • DG-Hist-Succ The synthesis of DG-Hist-Succ is effected in three steps. Initially, 3.7 mmol (2 g) of dipalmitoylglycerol is esterified with 4.1 mmol amino-protected CBZ-histidine. Ester formation is effected adding 4.3 mmol (0.67 g) of EDC and 4.3 mmol (0.53 g) of DMAP in 60 ml of dichloromethane. The batch is stirred for 4 hours. Deprotection of the histidine amino function furnishes the intermediate DG-Hist. The amino group is deprotected by means of catalytic hydrogenolysis on 10% palladium/carbon and stirring under hydrogen atmosphere overnight.
  • succinic anhydride is opened with benzyl alcohol to form benzyl succinate.
  • 10 mmol (1 g) of succinic anhydride and 9.5 mmol (1 g) of benzyl alcohol are dissolved in 50 ml of toluene.
  • 1 mmol (190 mg) of p-toluenesulfonic acid monohydrate this is heated at reflux for two hours.
  • 2 mmol (0.42 g) of benzyl-protected succinate is coupled to 1.6 mmol (1.13 g) DG-Hist via an amide bond.
  • the reaction is effected adding 2.5 mmol (0.39 g) of EDC and 2.5 mmol (0.3 g) of DMAP in 50 ml of dichloromethane, with stirring at room temperature for 4 hours. Finally, the succinate is deprotected by cleavage of the benzyl residue using catalytic hydrogenolysis on 10% palladium/carbon and stirring under hydrogen atmosphere overnight. The reaction batch is concentrated in vacuum, followed by purification using column chromatography on silica gel 60. Chloroform/methanol/ammonia (25% solution) 60:40:2 is used as eluent.
  • the amino group of 3-amino-1,2-propanediol is protected with tert-butyl acrylate.
  • 111 mmol (10 g) of 3-amino-1,2-propanediol is placed in 100 ml of acetonitrile.
  • 222 mmol (28.5 g) of tert-butyl acrylate is added under protective gas, and the batch is heated at reflux for two days.
  • the reaction mixture is concentrated in a rotary evaporator and purified using column chromatography on silica gel. Ethyl acetate/methanol 9:1 is used as eluent.
  • Unilamellar liposomes (DPPC/DPPG/cholesterol 40:20:40 mole-%) are suspended at a concentration of 20 mM lipid in a borate buffer (20 mM sodium borate, 120 mM sodium chloride, pH 8.4). 2 ml of this solution is added with 400 ⁇ l of a 0.6 M sodium periodate solution, and the mixture is incubated for 30 min in the dark. 1 ml of this suspension is chromatographed on Sephadex G25 in the borate buffer used above. The eluate of the liposome suspension is filled up to make 4 ml.
  • the liposomes thus oxidized are added with carnosine at a final concentration of 20 mM and incubated for 2 hours. Finally, this is reduced with 20 mM sodium borohydride at 4° C overnight. Excess carnosine can be removed by chromatography on Sephadex G25 as above.
  • the lipid film is hydrated directly with 1 ml DNA-containing (100 ⁇ g DNA/ml) NaAc buffer (10 mM NaAc, 150 mM NaCl, pH 4; slight ultrasonication, followed by rotation above the phase transition temperature for 30 min). This is followed by a freeze/thaw step.
  • the mixture is extruded 15 times through 400 nm membranes at a temperature 10° C. above the phase transition temperature.
  • Non-entrapped DNA can be removed by flotation in a sucrose gradient (at pH 7.5; 0.8 M sucrose, 0.5 M sucrose, buffer).
  • the DNA content is determined using the intercalation dye propidium iodide, an increase in fluorescence intensity occurring in case of intercalation into the DNA.
  • 20 ⁇ l of propidium iodide and 6 ⁇ l of Triton X-100 (10% in water) are filled up with sample to make 300 ⁇ l and measured using a fluorescent plate reader.

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Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1764090A1 (de) * 2005-09-15 2007-03-21 Novosom AG Amphotere Liposomen zur lokalen Verabreichung von Arzneistoffen
WO2008074488A2 (en) * 2006-12-19 2008-06-26 Novosom Ag Construction and use of transfection enhancer elements
WO2008151034A1 (en) * 2007-05-31 2008-12-11 The University Of Alabama Improved process for purification of aryl carboxylic acids
US20080311181A1 (en) * 2004-03-28 2008-12-18 Gerold Endert Serum-Stable Amphoteric Liposomes
US20100305198A1 (en) * 2007-10-19 2010-12-02 The University Court Of The University Of Edinburg Cationic lipids
WO2012109495A1 (en) 2011-02-09 2012-08-16 Metabolic Solutions Development Company, Llc Cellular targets of thiazolidinediones
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WO2014071406A1 (en) 2012-11-05 2014-05-08 Pronai Therapeutics, Inc. Methods of using biomarkers for the treatment of cancer by modulation of bcl2|expression
EP3000480A1 (de) 2005-12-01 2016-03-30 ProNAi Therapeutics, Inc. Krebstherapien und dabei verwendete pharmazeutische zusammensetzungen
US9795566B2 (en) 2014-07-17 2017-10-24 Fujifilm Corporation Imidazole compound and liposome containing same
US20180043036A1 (en) * 2015-02-04 2018-02-15 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. LIPID ASSEMBLIES AND USES THEREOF AND SOME pH AND ELECTROSTATIC MODULATING LIPIDS TO BE USED IN SAID ASSEMBLIES
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Publication number Priority date Publication date Assignee Title
DE10207178A1 (de) * 2002-02-19 2003-09-04 Novosom Ag Komponenten für die Herstellung amphoterer Liposomen
EP1549352A4 (de) * 2002-05-06 2005-07-27 Nucleonics Inc Verfahren zur abgabe von nukleinsäuren
DE102004056659A1 (de) * 2004-11-19 2006-06-01 Novosom Ag Abgabe von Oligonucleotiden an entzündete Schleimhaut
US9006487B2 (en) 2005-06-15 2015-04-14 Massachusetts Institute Of Technology Amine-containing lipids and uses thereof
EP1764091B1 (de) 2005-09-15 2017-08-30 Marina Biotech, Inc. Verbesserungen für, oder in Bezug auf amphotere Liposomen
US20080138393A1 (en) * 2006-12-11 2008-06-12 Access To Business Group International Llc Water soluble extract of spinach for prevention and repair of DNA damage
WO2008074487A2 (en) * 2006-12-19 2008-06-26 Novosom Ag Lipids and lipid assemblies comprising transfection enhancer elements
WO2008154700A1 (en) 2007-06-20 2008-12-24 Phylogica Limited Compositions and uses thereof for the treatment of acute respiratory distress syndrome (ards) and clinical disorders associated with therewith
US8969353B2 (en) 2008-11-07 2015-03-03 Massachusetts Institute Of Technology Aminoalcohol lipidoids and uses thereof
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US9193827B2 (en) 2010-08-26 2015-11-24 Massachusetts Institute Of Technology Poly(beta-amino alcohols), their preparation, and uses thereof
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EP2714971A4 (de) 2011-05-23 2015-01-21 Phylogica Ltd Verfahren zur bestimmung, identifizierung und isolierung zellpenetrierender peptide
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JP2013043885A (ja) * 2011-08-26 2013-03-04 Kansai Bunri Sogo Gakuen デヒドロアミノ酸含有グリセロール誘導体
US9464124B2 (en) 2011-09-12 2016-10-11 Moderna Therapeutics, Inc. Engineered nucleic acids and methods of use thereof
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ES2923757T3 (es) 2011-12-16 2022-09-30 Modernatx Inc Composiciones de ARNm modificado
US9303079B2 (en) 2012-04-02 2016-04-05 Moderna Therapeutics, Inc. Modified polynucleotides for the production of cytoplasmic and cytoskeletal proteins
AU2013243953A1 (en) 2012-04-02 2014-10-30 Modernatx, Inc. Modified polynucleotides for the production of nuclear proteins
US9572897B2 (en) 2012-04-02 2017-02-21 Modernatx, Inc. Modified polynucleotides for the production of cytoplasmic and cytoskeletal proteins
US9283287B2 (en) 2012-04-02 2016-03-15 Moderna Therapeutics, Inc. Modified polynucleotides for the production of nuclear proteins
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CA2884870C (en) 2012-08-13 2022-03-29 Massachusetts Institute Of Technology Amine-containing lipidoids and uses thereof
DK2922554T3 (en) 2012-11-26 2022-05-23 Modernatx Inc Terminalt modificeret rna
JP2016501887A (ja) * 2012-12-07 2016-01-21 カウンスィル オブ サイエンティフィック アンド インダストリアル リサーチＣｏｕｎｃｉｌ Ｏｆ Ｓｃｉｅｎｔｉｆｉｃ ＆ Ｉｎｄｕｓｔｒｉａｌ Ｒｅｓｅａｒｃｈ ヒスチジン官能性を有するカチオン性両親媒性化合物およびその製造プロセス、並びにリポソーム製剤
WO2014152211A1 (en) 2013-03-14 2014-09-25 Moderna Therapeutics, Inc. Formulation and delivery of modified nucleoside, nucleotide, and nucleic acid compositions
DK2970955T3 (en) 2013-03-14 2019-02-11 Translate Bio Inc METHODS FOR CLEANING MESSENGER RNA
KR20210122917A (ko) 2013-03-14 2021-10-12 샤이어 휴먼 지네틱 테라피즈 인크. Cftr mrna 조성물 및 관련 방법 및 사용
US8980864B2 (en) 2013-03-15 2015-03-17 Moderna Therapeutics, Inc. Compositions and methods of altering cholesterol levels
US9315472B2 (en) 2013-05-01 2016-04-19 Massachusetts Institute Of Technology 1,3,5-triazinane-2,4,6-trione derivatives and uses thereof
JP5914418B2 (ja) 2013-06-26 2016-05-11 富士フイルム株式会社 脂質粒子、核酸送達キャリア、核酸送達キャリア製造用組成物、脂質粒子の製造方法及び遺伝子導入方法
WO2015048744A2 (en) 2013-09-30 2015-04-02 Moderna Therapeutics, Inc. Polynucleotides encoding immune modulating polypeptides
WO2015051214A1 (en) 2013-10-03 2015-04-09 Moderna Therapeutics, Inc. Polynucleotides encoding low density lipoprotein receptor
PE20161242A1 (es) 2013-10-22 2016-12-11 Massachusetts Inst Technology Formulaciones de lipidos para la administracion de arn mensajero
EA034103B1 (ru) 2013-10-22 2019-12-27 Транслейт Био, Инк. СПОСОБ ЛЕЧЕНИЯ ФЕНИЛКЕТОНУРИИ С ПРИМЕНЕНИЕМ мРНК
US11224642B2 (en) 2013-10-22 2022-01-18 Translate Bio, Inc. MRNA therapy for argininosuccinate synthetase deficiency
DK3071696T3 (da) 2013-11-22 2019-10-07 Mina Therapeutics Ltd C/ebp alfa kort aktiverings-rna-sammensætninger og fremgangsmåder til anvendelse
EP3134506B1 (de) 2014-04-25 2019-08-07 Translate Bio, Inc. Verfahren zur reinigung von messenger-rna
BR112016027705A2 (pt) 2014-05-30 2018-01-30 Shire Human Genetic Therapies lipídios biodegradáveis para distribuição de ácidos nucleicos
UA121863C2 (uk) 2014-06-24 2020-08-10 Транслейт Байо, Інк. Стереохімічно збагачені композиції для доставки нуклеїнових кислот
WO2016004202A1 (en) 2014-07-02 2016-01-07 Massachusetts Institute Of Technology Polyamine-fatty acid derived lipidoids and uses thereof
JP6240570B2 (ja) 2014-07-17 2017-11-29 富士フイルム株式会社 脂質粒子および核酸送達キャリア
RS64331B1 (sr) 2015-06-19 2023-08-31 Massachusetts Inst Technology Alkenil supstituisani 2,5-piperazindioni i njihova primena u sastavima za isporuku agensa u organizam ili ćeliju subjekta
WO2017031232A1 (en) 2015-08-17 2017-02-23 Modernatx, Inc. Methods for preparing particles and related compositions
LT3350157T (lt) 2015-09-17 2022-02-25 Modernatx, Inc. Junginiai ir kompozicijos terapinei medžiagai teikti intraceliuliniu būdu
AU2016366978B2 (en) 2015-12-10 2022-07-28 Modernatx, Inc. Compositions and methods for delivery of therapeutic agents
EP4036079A3 (de) 2015-12-22 2022-09-28 ModernaTX, Inc. Verbindungen und zusammensetzungen zur intrazellulären verabreichung von wirkstoffen
EP3538067A1 (de) 2016-11-08 2019-09-18 Modernatx, Inc. Stabilisierte formulierungen von lipidnanopartikeln
AU2018224326B2 (en) 2017-02-27 2024-01-04 Translate Bio, Inc. Novel codon-optimized CFTR mRNA
EP3595727A1 (de) 2017-03-15 2020-01-22 ModernaTX, Inc. Lipidnanopartikelformulierung
CN110520409A (zh) 2017-03-15 2019-11-29 摩登纳特斯有限公司 用于细胞内递送治疗剂的化合物和组合物
DK3596042T3 (da) 2017-03-15 2022-04-11 Modernatx Inc Krystalformer af aminolipider
JOP20190260A1 (ar) 2017-05-02 2019-10-31 Merck Sharp & Dohme صيغ ثابتة لأجسام مضادة لمستقبل الموت المبرمج 1 (pd-1) وطرق استخدامها
RU2019138507A (ru) 2017-05-02 2021-06-02 Мерк Шарп И Доум Корп. Составы антител против lag3 и совместные составы антител против lag3 и антител против pd-1
EP3624824A1 (de) 2017-05-16 2020-03-25 Translate Bio, Inc. Behandlung von zystischer fibrose durch verabreichung von codonoptimierter mrna, die cftr codiert
EP3679140B1 (de) 2017-09-08 2022-11-16 MiNA Therapeutics Limited Stabilisierte cebpa-sarna-zusammensetzungen und verfahren zur verwendung
AU2018330495A1 (en) 2017-09-08 2020-03-26 Mina Therapeutics Limited Stabilized hnf4a sarna compositions and methods of use
EP3775211B1 (de) 2018-04-12 2023-04-05 MiNA Therapeutics Limited Sirt1-sarna kompositionen und methoden zu deren verwendung
CA3108544A1 (en) 2018-08-24 2020-02-27 Translate Bio, Inc. Methods for purification of messenger rna
WO2020208361A1 (en) 2019-04-12 2020-10-15 Mina Therapeutics Limited Sirt1-sarna compositions and methods of use
CN114728887A (zh) 2019-09-19 2022-07-08 摩登纳特斯有限公司 用于治疗剂的细胞内递送的支链尾端脂质化合物和组合物
GB2603454A (en) 2020-12-09 2022-08-10 Ucl Business Ltd Novel therapeutics for the treatment of neurodegenerative disorders
CN114685778B (zh) * 2020-12-30 2023-10-17 苏州艾博生物科技有限公司 长循环阳离子脂质体的合成方法
US11524023B2 (en) 2021-02-19 2022-12-13 Modernatx, Inc. Lipid nanoparticle compositions and methods of formulating the same
AU2022246144A1 (en) 2021-03-26 2023-09-21 Mina Therapeutics Limited Tmem173 sarna compositions and methods of use
WO2023099884A1 (en) 2021-12-01 2023-06-08 Mina Therapeutics Limited Pax6 sarna compositions and methods of use
GB202117758D0 (en) 2021-12-09 2022-01-26 Ucl Business Ltd Therapeutics for the treatment of neurodegenerative disorders
WO2023170435A1 (en) 2022-03-07 2023-09-14 Mina Therapeutics Limited Il10 sarna compositions and methods of use

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4788182A (en) * 1984-07-25 1988-11-29 Ciba-Geigy Corporation Phosphatidyl compounds, processes for their manufacture, and their use
US6022720A (en) * 1997-07-17 2000-02-08 Glaxo Wellcome Inc. Bax protein channel formation
US6043094A (en) * 1996-10-11 2000-03-28 Sequus Pharmaceuticals, Inc. Therapeutic liposome composition and method
US6090800A (en) * 1997-05-06 2000-07-18 Imarx Pharmaceutical Corp. Lipid soluble steroid prodrugs
US6180134B1 (en) * 1993-03-23 2001-01-30 Sequus Pharmaceuticals, Inc. Enhanced ciruclation effector composition and method
US20010033862A1 (en) * 1988-07-07 2001-10-25 Hostetler Karl Y. Methods of treating viral infections using antiviral liponucleotides
US6379698B1 (en) * 1999-04-06 2002-04-30 Isis Pharmaceuticals, Inc. Fusogenic lipids and vesicles

Family Cites Families (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2032259A1 (en) * 1989-12-18 1991-06-19 Wayne J. Thompson Hiv protease inhibitors useful for the treatment of aids
JP2604268B2 (ja) * 1990-04-09 1997-04-30 富士写真フイルム株式会社 ペプチド誘導体両親媒性化合物、その中間体、ペプチド誘導体両親媒性化合物を用いたリポソームおよび薄膜
US5206027A (en) * 1990-09-13 1993-04-27 Fuji Photo Film Co., Ltd. Amphipathic compound and liposome comprising the same
JP2597927B2 (ja) * 1990-11-29 1997-04-09 富士写真フイルム株式会社 リン脂質誘導体およびそれを用いたリポソーム
US5334761A (en) * 1992-08-28 1994-08-02 Life Technologies, Inc. Cationic lipids
JP2886748B2 (ja) 1992-09-17 1999-04-26 富士写真フイルム株式会社 写真用処理組成物及び処理方法
CA2194221C (en) * 1994-06-22 2001-01-02 Timothy D. Heath Cationic amphiphiles
ES1045340Y (es) * 2000-02-11 2001-02-16 Luque Fernandez Antonio De Cuña elevadora para camas de accionamiento neumatico.
DE10109898A1 (de) * 2001-02-21 2002-09-05 Novosom Gmbh Lipide mit veränderlicher Ladung
GB0106041D0 (en) * 2001-03-12 2001-05-02 Cancer Res Ventures Ltd Lipids and liposomes
DE10207178A1 (de) * 2002-02-19 2003-09-04 Novosom Ag Komponenten für die Herstellung amphoterer Liposomen

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4788182A (en) * 1984-07-25 1988-11-29 Ciba-Geigy Corporation Phosphatidyl compounds, processes for their manufacture, and their use
US20010033862A1 (en) * 1988-07-07 2001-10-25 Hostetler Karl Y. Methods of treating viral infections using antiviral liponucleotides
US6180134B1 (en) * 1993-03-23 2001-01-30 Sequus Pharmaceuticals, Inc. Enhanced ciruclation effector composition and method
US6043094A (en) * 1996-10-11 2000-03-28 Sequus Pharmaceuticals, Inc. Therapeutic liposome composition and method
US6090800A (en) * 1997-05-06 2000-07-18 Imarx Pharmaceutical Corp. Lipid soluble steroid prodrugs
US6022720A (en) * 1997-07-17 2000-02-08 Glaxo Wellcome Inc. Bax protein channel formation
US6379698B1 (en) * 1999-04-06 2002-04-30 Isis Pharmaceuticals, Inc. Fusogenic lipids and vesicles

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8236770B2 (en) 2004-03-28 2012-08-07 Marina Biotech, Inc. Serum-stable amphoteric liposomes
US20080311181A1 (en) * 2004-03-28 2008-12-18 Gerold Endert Serum-Stable Amphoteric Liposomes
EP1764090A1 (de) * 2005-09-15 2007-03-21 Novosom AG Amphotere Liposomen zur lokalen Verabreichung von Arzneistoffen
EP3000480A1 (de) 2005-12-01 2016-03-30 ProNAi Therapeutics, Inc. Krebstherapien und dabei verwendete pharmazeutische zusammensetzungen
WO2008074488A2 (en) * 2006-12-19 2008-06-26 Novosom Ag Construction and use of transfection enhancer elements
WO2008074488A3 (en) * 2006-12-19 2009-03-26 Novosom Ag Construction and use of transfection enhancer elements
US20100022621A1 (en) * 2006-12-19 2010-01-28 Steffen Panzner Construction and use of transfection enhancer elements
US8309760B2 (en) 2007-05-31 2012-11-13 University Of Alabama Process for purification of aryl carboxylic acids
US20100174111A1 (en) * 2007-05-31 2010-07-08 Rogers Robin D Process for purification of aryl carboxylic acids
WO2008151034A1 (en) * 2007-05-31 2008-12-11 The University Of Alabama Improved process for purification of aryl carboxylic acids
US8859806B2 (en) 2007-05-31 2014-10-14 University Of Alabama Process for purification of aryl carboxylic acids
US20100305198A1 (en) * 2007-10-19 2010-12-02 The University Court Of The University Of Edinburg Cationic lipids
WO2012109495A1 (en) 2011-02-09 2012-08-16 Metabolic Solutions Development Company, Llc Cellular targets of thiazolidinediones
CN103596922A (zh) * 2011-06-07 2014-02-19 因塞拉有限公司 氨基脂质，及其合成和用途
US8759464B2 (en) 2011-10-05 2014-06-24 Arkema France Polar, multi-hydroxyl functional amino compounds, compositions, process for preparation, their uses and applications
WO2013050487A1 (en) * 2011-10-05 2013-04-11 Arkema France Polyhydroxyl - substituted amino compounds, polymers containing, and their use
WO2014071406A1 (en) 2012-11-05 2014-05-08 Pronai Therapeutics, Inc. Methods of using biomarkers for the treatment of cancer by modulation of bcl2|expression
US9795566B2 (en) 2014-07-17 2017-10-24 Fujifilm Corporation Imidazole compound and liposome containing same
US20180043036A1 (en) * 2015-02-04 2018-02-15 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. LIPID ASSEMBLIES AND USES THEREOF AND SOME pH AND ELECTROSTATIC MODULATING LIPIDS TO BE USED IN SAID ASSEMBLIES
US10722599B2 (en) * 2015-02-04 2020-07-28 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Lipid assemblies and uses thereof and some pH and electrostatic modulating lipids to be used in said assemblies
CN115463093A (zh) * 2022-09-28 2022-12-13 天津市人民医院 一种奥沙利铂制剂及其制备方法

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US9668973B2 (en) 2017-06-06
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