EP1480621A1 - Ph sensitiv kationische lipide, lipsomen und nanokapseln, die diese umfassen - Google Patents

Ph sensitiv kationische lipide, lipsomen und nanokapseln, die diese umfassen

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Publication number
EP1480621A1
EP1480621A1 EP03742540A EP03742540A EP1480621A1 EP 1480621 A1 EP1480621 A1 EP 1480621A1 EP 03742540 A EP03742540 A EP 03742540A EP 03742540 A EP03742540 A EP 03742540A EP 1480621 A1 EP1480621 A1 EP 1480621A1
Authority
EP
European Patent Office
Prior art keywords
liposomes
lipid
acid
lipids
nanocapsules
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
EP03742540A
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German (de)
English (en)
French (fr)
Inventor
Frank Essler
Steffen Panzner
Gerold Endert
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Novosom AG
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Novosom AG
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Filing date
Publication date
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Publication of EP1480621A1 publication Critical patent/EP1480621A1/de
Ceased legal-status Critical Current

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    • 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
    • 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
    • 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
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/10—Antimycotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • 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
    • C07D295/04—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 with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/08—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 with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms
    • C07D295/084—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 with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms with the ring nitrogen atoms and the oxygen or sulfur atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings
    • C07D295/088—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 with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms with the ring nitrogen atoms and the oxygen or sulfur atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings to an acyclic saturated chain
    • 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
    • C07D295/04—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 with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/12—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 with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms
    • C07D295/125—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 with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms with the ring nitrogen atoms and the substituent nitrogen atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings
    • C07D295/13—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 with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms with the ring nitrogen atoms and the substituent nitrogen atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings to an acyclic saturated chain

Definitions

  • the invention relates to pH sensitive, cationic lipids, i.e. polar compounds based on amphiphilic molecules, 0 where one or more organic cations with a pK value between 3, 5 and 8 are substituted on their hydrophilic head group; the invention further relates to liposomes and nanocapsules containing these compounds and the use of these liposomes. 5
  • lipids summarizes three classes of natural products that can be isolated from biological membranes: .phospholipids, sphingolipids and cholesterol with its derivatives. Technically manufactured connections with similar properties are e.g. the diacylglycerides or N, N-dialkylamines.
  • liposomes can be used 5 among other things as containers for active substances in pharmaceutical preparations. Efficient and stable packaging of the cargo and controllable release of the contents are desirable. Both / claims cannot be easily combined: the more stable the 0 and the denser the packaging, the more difficult it is to release the enclosed active ingredient. For this reason, liposomes have been developed that change their properties in response to an external stimulus, such as concentration, temperature or pH. 5 In the prior art, various thermosensitive and pH-sensitive liposomes known. The pH-sensitive liposomes are of particular interest because this parameter can also change under physiological circumstances, for example when a liposome is endocytosed in cells or when the gastrointestinal tract is passed.
  • Known pH-sensitive liposomes include CHEMS in particular.
  • CHEMS used in a mixture with phosphatidylethanolamine for the preparation of pH-sensitive liposomes (Tachibana et al. (1998); BBRC 251: 538-544, US4891208).
  • Such liposomes can be endocytosed by cells and in this way are able to transport cargo molecules into the interior of cells without violating the integrity of the cellular membrane.
  • a major disadvantage of the CHEMS is its anionic character.
  • the liposomes thus produced have a negative total charge and are disadvantageously taken up by cells with little efficiency. Despite the transfer mechanism described above, they are therefore hardly suitable for the introduction of macromolecules into cells.
  • cationic liposomes are used which have the highest possible and constant surface charge.
  • the positive total charge of such particles leads to electrostatic attachment to cells and consequently to efficient transport.
  • the use of these connections and thus, the liposomes produced remain restricted to applications in vitro or ex vivo, since such positively charged liposomes with serum components disadvantageously form uncontrolled aggregates.
  • the disadvantages of the known lipids and the structures that are formed from these lipids can be summarized as follows:
  • the amount of proteins or DNA or RNA that can be included in anionic liposomes is generally below average and is not sufficient for a wide variety of applications out .
  • a suitable amount of proteins or nucleic acids or their derivatives can be included by using known cationic liposomes, but these structures often have a critical cytotoxicity.
  • i) with the help of active ingredients can be stably enclosed in liposomes, ii) with the help of which liposomes can be produced which can transport an enclosed active ingredient into the interior of cells, iii) the presence of which enables the production of cationic liposomes without formation larger aggregates can be mixed with serum, iv) which can be incorporated in a high proportion in liposomal membranes and v) which have good biodegradability with sufficient biological stability.
  • the invention solves this technical problem by providing a pH-sensitive, cationic lipid with a pKa value between 3, 5 and 8 according to the general formula (I):
  • the cation is selected from the group comprising imidazole, morpholine, piperazine, purine, pyridine and / or pyrimidine or a derivative thereof,
  • amphiphile comprises a structure of the general formula (II) or (III):
  • Rl and R2 are independently C8 to C30 alkyl or acyl chains with 0, 1 or 2 ethylenically unsaturated bonds and
  • Rl and R2 are independently C8 to C30 acyl chains with 0, 1 or 2 ethylenically unsaturated bonds and X is -O-.
  • amphiphile comprises both membrane-forming and membrane-bound connections of a bilayer membrane.
  • Preferred building blocks of the compounds according to the invention are amphiphiles such as diacylglycerides, dialkylglycerides, phosphoglycerides, acylated or alkylated 3-amino-1,2-propanediols, since these compounds are particularly cheaply available, have simple chemistry and can be incorporated to a large extent in membranes, without increasing their permeability or even destroying the membrane character, preferably destroying them completely.
  • amphiphiles are also preferred: diacylglycerols, dialkylglycerols, phosphoglycerols, acylated or alkylated 3-amino-1,2-propanediols.
  • the long-chain alkyls or acyls present in these fragments contain between 8 and 30 carbon atoms. They are preferably straight-chain or little branched and can have 0, 1 or 2 ethylenically unsaturated bonds. Substituents which are found in natural lipids, ie straight-chain fatty acids or fatty alcohols having 12 to 20 carbon atoms and none, one or two unsaturated bonds, are particularly preferred.
  • Lauroyl, myristoyl, palmitoyl, stearoyl, oleoyl and linoyl residues and their corresponding alcohols are very particularly preferred.
  • dicarboxylic acids are used as the polar head group of the amphiphile, which in particular allow coupling of the ultimately charge-bearing substituents via further functional groups.
  • the following amphiphiles derived therefrom are preferred: long-chain esters of 1,4- or 1,5- Dicarboxylic acids, especially aspartic acid, glutamic acid, malic acid, tartaric acid, citric acid, aconitic acid, citraconic acid, maleic acid or similar compounds in which the substituents which ultimately bear the charge are coupled via the remaining amino group, hydroxyl group, carboxyl group or via the double bond.
  • amphiphiles are obtained from diamines with a further functional group, for example as the diamide of 3-aminoalanine, diaminobutyric acid, ornithine or lysine with long-chain fatty acids.
  • the compounds of the invention can also be prepared as derivatives of sphingosine or ceramides. They can also be represented as derivatives of long-chain vinyl ethers or plasmalogens.
  • amphiphiles which are advantageously used as the starting material, can be functionalized differently on their hydrophilic head group in order expediently to allow simple and permanent coupling or optionally to perform the function of a spatial spacer.
  • the hydroxyl group, the amino group or else carboxyl groups are particularly suitable for direct coupling. These coupling groups are advantageously readily biodegradable.
  • Y and X represent connecting functional groups, the presence of which is not mandatory in every case; it can be provided that the value for Y and / or X is a
  • the connecting group X preferably comprises the structure
  • Group Y can be omitted, for example, if the cation can be coupled directly to the amphiphile, for example in the esterification of imidazole acetic acid with dipalmitoylglycerol.
  • the molecular building blocks are between the cation and the amphiphile: Spacer - Y - Spacer - X.
  • the spacer is a lower alkyl radical with a linear, branched or ring-shaped structure, which has 0 to 8 C atoms and 0, 1 or 2 ethylenically unsaturated Contains bonds, preferably -CH 2 - ; -CH 2 -CH 2 - and / or -CH 2 -CH 2 -CH 2 -.
  • the spacer can have hydroxyl groups to increase the polarity of the molecule.
  • the spacer can in particular be a sugar.
  • the spacer can also advantageously be a polyethylene glycol, with such a spacer comprising up to 20 monomer units.
  • the whole molecule receives its pH-dependent charge characteristic from one or more organic cations with a pKa value between 3, 5 and 8.
  • Preferred molecules or molecular building blocks with this property are nitrogen bases. These nitrogen bases are linked to the lipid via spacers and coupling groups, so that a compound is formed according to the formula according to the invention.
  • Toluidines or phenetidines 2-, 3-, 5-, 6-, 7- or 8-substituted benzimidazoles, 2-, 3, 4- or 5-substituted imidazoles, 1-, or 5-substituted isoquinolines, 2-, 3- or 4-substituted morpholines, 2-, 3- or 4-substituted picolines, 1-, 2-, or 3-substituted piperazines, 2-, 5- or 6-modified pterins, 3-, 4-, 5-, 6 - or 9-substituted purines, 2- or 3-substituted pyrazines, 3- or 4-substituted pyridazines, 2-, 3- or 4-substituted pyridines, 2-, 4-, 5- or 6-substituted pyrimidines, l- , 2-, 3-, 4-, 5-, 6- or 8-substituted quinolines, 2-, 4- or 5- substituted thiazoles, 2-, 4- or 6- substituted triaz
  • positional isomers in which the connecting spacer is substituted at different positions on the organic cation.
  • Such positional isomers are within the scope of the disclosure of this invention.
  • the position isomerism alone can influence the pKa values of the organic cations.
  • the underlying rules are familiar to the person skilled in the art. Alternatively, these influences can be estimated from tables (Handbook of Chemistry and Physics, 73rd volume, pp. 8-37 ff.).
  • Rl and / or R2 are preferably independently of one another C8 to C30 alkyl or alkyl chains with 0, 1 or 2 ethylenically unsaturated bonds, in particular lauroyl, myristoyl, palmitoyl, stearoyl, oleoyl and linoyl radicals, or their corresponding alcohols.
  • the person skilled in the art is aware that in addition to the preferred amphiphiles, Y, X, R1, R2, spacers and / or cations, he can also use further amphiphiles, Y, X, R1, R2 spacers and / or cations; In such a case, the preferred amphiphiles Y, X, R1, R2, spacers and / or cations are not an essential component of the structures according to the invention. It can furthermore be advantageous to dispense with the spacer, Y and / or X.
  • the compound has a pKa value between 3.5 and 7, preferably between 4 and 6.5. This pKa value is advantageously in a range which is of crucial importance for the physiology of numerous organisms.
  • the cations are derivatives of piperazines, imidazoles, morpholines, purines and / or pyrimidines.
  • Molecular fragments such as those found in biological systems are very particularly preferred, in particular 4-imidazoles (histamines), 2, 6 or 9 purines (adenines, guanines, adenosines or guanosines), 1, 2 or 4 pyrimidines (Uracile, thymine, cytosine, uridine, thymidine, cytidine) or also pyridine-3-carboxylic acids (nicotinic acid esters or amides).
  • the structural fragments mentioned here can of course have further substituents. These can be, for example, methyl, ethyl, propyl or isopropyl radicals, particularly preferably in hydroxylated form with one or two hydroxyl groups. However, this can also be hydroxyl or keto functions of the ring system.
  • Nitrogen bases with preferred pKa values are also created by single or multiple substitution of the Nitrogen atom with Niederalkanhydroxylen, such as hydroxymethyl or hydroxyethyl groups.
  • Suitable organic bases from this group are, for example, aminopropanediols, triethanolamines, tris (hydroxymethyl) methylamines, bis (hydroxymethyl) methylamines, tris (hydroxyethyl) methylamines, bis (hydroxyethyl) methylamines or the correspondingly substituted ethylamines.
  • Nitrogen bases with preferred pKa values can also be found among the amino sugars or amino sugar alcohols. These fragments can be coupled to the hydrophobic part of the molecule either via the nitrogen of the base or via one of the hydroxyl functions.
  • a suitable complex group is the amide from histamine and histidine or from histamine and histindinylhistidine.
  • Anionic groups such as carboxylic acids, sulfonic acids, enols or aromatic hydroxyls may only be part of the molecule if they are not dissociated in the claimed pH range between 3.5 'and 8.5. This is generally the case if their pKa is above 9.5.
  • amphiphile is selected from
  • Rl and R2 are independently C8 to C30 alkyl with 0, 1 or 2 ethylenically unsaturated bonds
  • X is selected from the group consisting of a deletion; -0-; -NH- and / or -S- ;.
  • Synthesis methods Methods for chemical coupling of the individual molecular building blocks are known to the person skilled in the art and can vary depending on the starting material and coupling component used. Typical reactions are esterification, amidation, the addition of amines to double bonds, etherification or reductive amination.
  • Particularly preferred molecules can be produced by:
  • Diacylglycerol hemimaleats iv) amidation of phosphatidylethanolamine or phosphatidylserine, v) amidation or alkylation of 3-amino-1,2-propanediol diesters, vi) oxidation of phosphatidylglycerols and subsequent reductive amination and vii) reductive amination of glyceraldehyde and subsequent acylation.
  • the coupling is preferably carried out of an amphiphile, which has a free carboxyl function as an amide. Amidation of is also preferred
  • the radicals Rl and R2 are acyl, alkyl or alkenyl groups.
  • the coupling is advantageously carried out after the oxidation of the terminal glycerol to the aldehyde and subsequent reductive coupling of the amphoteric, in this example with histamine.
  • the radicals R are alkyl or alkenyl groups.
  • radicals R are alkyl or alkenyl groups.
  • N- (Aspartyldihydroxyalkyl) -N '- histaminylurea The radicals R are alkyl or alkenyl groups.
  • radicals R are alkyl or alkenyl groups.
  • Glucosamine on glyceraldehyde and subsequent acylation are acyl, alkyl or
  • Aminobenzoic acid or diaminobenzoic acid derivatives are preferably carried out as an ester or amide of benzoic acid.
  • the lipid can be, for example, a 3-amino-1, 2-propanediol derivative.
  • the addition can be, for example, that a further cation, a further spacer, a further X, a further Y and / or a further amphiphile are added to the lipids according to the invention;
  • a further cation, a further spacer, a further X, a further Y and / or a further amphiphile are added to the lipids according to the invention;
  • several other cations, spacers, X, Y and / or amphiphiles are added to a lipid according to the invention.
  • individual areas of the lipid or the entire lipid it is possible for individual areas of the lipid or the entire lipid to be duplicated at least once. It is also known to the person skilled in the art that he can replace individual constituents of the lipids according to the invention with constituents having a similar effect.
  • lipids according to the invention it is possible to change the structure of the lipids according to the invention in such a way that • a specific sequence within the lipid or within a partial region of the lipid is changed so that, for example, the cation is bound directly to the amphiphile and via the X or the Y to the amphiphile spacer is present bound, such compounds are essentially functionally analogous to the lipids according to the invention in the sense of the invention. It is also possible to remove an entire area from the lipid and reinstall it in reverse.
  • the chemist is aware that almost all modification and mutation possibilities that exist, for example, in the field of peptide or protein or nucleic acid chemistry, can also be used in an analogous manner in the field of lipid chemistry.
  • the invention also relates to liposomes comprising the substances according to the invention. All substances or compounds according to the invention can advantageously be incorporated in a high proportion in liposomal membranes and only lead to a positive charge of the entire particle when the pH of the medium is less than (pKa + 1) of the compounds according to the invention.
  • the proportion of the pH-sensitive cationic lipid is at most 50 mol%, in particular 40 mol% and preferably 30 mol%.
  • Compositions which contain at least 5 mol%, preferably 7 mol%, particularly preferably 10 mol%, very particularly preferably 15 mol%, but at most 40 mol% of the compound are particularly preferred.
  • Compositions which contain at least 10 mol% and at most 30 mol% of the pH-sensitive cationic lipid are very particularly preferred.
  • the liposomes comprise, in particular, phosphatidylcholine, phosphatidylethanolamine and / or diacylglycerol. Since cholesterols cannot form liposomes themselves, the addition of another lipid is necessary. This lipid can in particular be a phospholipid. Further modifications of the liposome are of course possible. The use of polyethylene glycol-modified phospholipids or analogous compounds is particularly advantageous.
  • anionic lipids such as phosphatidylglycerol, phosphatidylserine, Phosphatidic acid or CHEMS can be added.
  • the proportion of anionic lipids must not exceed that of the pH-sensitive cationic ones; it must be much smaller.
  • the liposomes have an average size between 50 and 1000 nm, in particular 50 and 500 nm, preferably between 50 and 300 nm and very particularly preferably between 60 and 130 nm.
  • the liposomes comprise water-soluble active ingredients.
  • the liposomes according to the invention are particularly suitable for parenteral use. They can preferably be used in cancer therapy and for the therapy of severe infections. Liposome dispersions can be injected, infused or implanted. They then distribute themselves in the blood or in the lymph or give off their active ingredient in a controlled manner as a depot. The latter can be achieved by highly concentrated dispersions, which are available as gels.
  • the liposomes can also be used for topical application on the skin. In particular, they can contribute to the fact that various active substances can penetrate the skin better or can particularly preferably get into the body through the skin. It is also possible to use the liposomes for gene transfer.
  • Liposomes which are produced using the substances according to the invention advantageously show only a low non-specific binding to cell surfaces. This is particularly the case if additional anionic lipids are used for the production. This low non-specific binding is an essential prerequisite for the establishment of specific binding to target cells. If the described liposomes are provided with further ligands, target control of the vehicle is given. The active substance can then be specifically enriched in cells or tissues which have pathological conditions.
  • An essential use of the substances according to the invention therefore lies in the construction of vectors for the transfer of active substances in living organisms.
  • the vectors are particularly suitable for the transport of therapeutic macromolecules, such as proteins or DNA, which cannot inherently cross the cell membrane or are rapidly broken down in the bloodstream.
  • the liposomes comprise as active ingredient a protein, a peptide, a carbohydrate structure, a DNA, an RNA, an antisense nucleotide and / or a decoy nucleotide.
  • the invention also relates to nanocapsules containing the lipids according to the invention and / or those according to the invention Include liposomes.
  • lipids according to the invention and / or those according to the invention Include liposomes.
  • Various possibilities are known to the person skilled in the art to provide liposomal nanocapsules, for example WO 00/28972 and / or WO 01/64330 disclose various methods for generating nanocapsules which are included in the disclosure content of the present application.
  • the invention also relates to the lipids according to the invention, the liposomes according to the invention and / or the nanocapsules according to the invention for use as a medicinal active substance.
  • the therapeutically active compounds are preferably not restricted to specific indications.
  • the active ingredient is an active ingredient for prophylaxis, diagnosis, therapy, follow-up and / or post-treatment, in particular of diseases.
  • it can be provided to load the lipids, liposomes and / or nanocapsules that are used as the active ingredient with a second active ingredient; which are used synonymously below.
  • the active ingredient is an active ingredient for prophylaxis, it can in particular be substances that are suitable, for example, for vaccination.
  • the lipids, the liposomes and / or the nanocapsules are used in vivo, in vitro or ex vivo in order to detect pathogenic modifications of an organism or a tissue culture or another isolated body component.
  • the healing of diseases and the restoration of the healthy Initial state or the production of the non-pathogenic initial state is sought, the absence of a certain disease does not mean that the organism can have no other pathogenic change. That is, when the structure according to the invention is used in the treatment and cure of a viral disease, the organism can also comprise a tumor disease which is initially not treated in it. Of course, it can also be provided that several diseases are treated simultaneously or with a time delay by the structures according to the invention.
  • the progress check can be, for example, a diagnosis that is carried out in parallel with a therapy at several time intervals. This can in particular be a treatment of metabolic diseases, cancer diseases, immunological diseases or genetic diseases.
  • the success of the therapy is then monitored and monitored with the help of the lipids, liposomes or nanocapsules used in the follow-up. Post-treatment includes, in particular, diagnostic and therapeutic actions after which a healing success has partially or essentially already occurred.
  • the invention also relates to a pharmaceutical composition
  • a pharmaceutical composition comprising at least one lipid according to the invention, at least one liposome according to the invention and / or a nanocapsule according to the invention, optionally together with a pharmaceutically acceptable carrier.
  • the structures according to the invention are used with or without an active ingredient as a pharmaceutical composition.
  • the pharmaceutical composition can be used in particular as a pharmaceutical.
  • the liposomes according to the invention are not attacked by complement components or perforin and can thus be used to transport active ingredients.
  • Medicaments or pharmaceutical compositions which are used synonymously in the present case are substances and preparations made of substances, in particular lipids, liposomes and / or nanocapsules, which are intended to cure diseases, ailments, physical damage or pathological complaints by application to or in the human body to alleviate or prevent.
  • medical auxiliaries or carriers are substances which are used for production as active ingredients of pharmaceuticals.
  • Pharmaceutical-technical auxiliaries serve for the suitable formulation of the medicament or the pharmaceutical composition and can even be subsequently removed, if they are only required during the production process, or can be part of the pharmaceutical composition as a pharmaceutically acceptable carrier.
  • the pharmaceutical formulation or formulation of the pharmaceutical composition is optionally carried out in combination with a pharmaceutically acceptable carrier and / or diluent.
  • Suitable pharmaceutically acceptable carriers include phosphate buffered saline, water, emulsions, gels, sterile solutions, etc.
  • Drugs or pharmaceutical compositions comprising such carriers can be formulated using known conventional methods.
  • This drug or pharmaceutical Compositions can be administered to an individual in a suitable dose, for example in a range from I ⁇ g to 10 g of lipids, liposomes and / or nanocapsules with and / or without an included active ingredient per day and patient.
  • Doses of 1 mg to 1 g are preferred, which can relate both to the lipids, liposomes, nanocapsules and / or the active ingredient transported through them.
  • Administration of as few and low doses as possible is preferred, and a single or multiple dose is further preferred.
  • the administration can be carried out in various ways, for example intravenously, intraperitoneally, intrarectally, intragastrointestinally, intranodally, intramuscularly, locally, for example in the tumor, but also subcutaneously, intradermally or on the skin or via the mucous membranes.
  • the administration can also take place in the form of gene therapies, the lipids, liposomes and / or nanocapsules being used to transport nucleic acids.
  • compositions or the medicament in particular comprise a pharmacological substance which is enclosed or bound by the lipids, liposomes and / or nanocapsules.
  • a pharmacological substance which is enclosed or bound by the lipids, liposomes and / or nanocapsules.
  • These can either be used alone with the corresponding auxiliaries described under drugs or pharmaceutical compositions or in combination with one or more adjuvants, for example QS-21, GPI-0100 or emulsions such as, for example, montanides, adjuvants, DNA compounds such as, for example, CpG, detox, bacterial vaccines , Salts such as calcium phosphates and / or another suitable substance for enhancing the effect are administered; preferably immunostimulatory molecules, such as interleukins, for example IL-2, IL-12, IL-4 and / or Growth factors, for example GM-CSF.
  • adjuvants for example QS-21, GPI-0100
  • emulsions such as, for example, mont
  • the pharmaceutical composition or the medicament can of course also be a combination of two or more of the pharmaceutical compositions or medicaments according to the invention, as well as a combination with other medicaments, such as, for example, antibody therapies, chemotherapy or radiotherapy, which are administered in a suitable manner over time or separately or be applied.
  • the pharmaceuticals or pharmaceutical compositions are produced by methods known per se.
  • the invention also relates to a kit comprising at least one lipid according to the invention, at least one liposome according to the invention, at least one nanocapsule according to the invention and / or a pharmaceutical composition according to the invention, optionally with information for combining the contents of the kit and / or for providing a formulation and the algorithm the administration of the formulation, ie at which dose or time intervals, in particular, individual components of the kit are administered to a patient.
  • the recipient in the sense of the invention can also be a cell or a tissue in vivo, ex vivo or in vitro.
  • the information can be, for example, an instruction leaflet, but also information that the user can call up by telephone or via the Internet.
  • the algorithm of the administration of the formulation contains, in particular, instructions for diagnostic and / or therapeutic method of treating a patient. These can be one-step or multi-step procedures, as well as those that are carried out in the absence or presence of a doctor. This means that the therapy scheme or the information about it can be part of the kit.
  • the invention also relates to the use of the lipids according to the invention, the liposomes according to the invention, the nanocapsules according to the invention, the kit according to the invention and / or for the manufacture of a medicament for the treatment of diseases, in particular for the treatment of genetic, immunological, metabolic or cell growth disorders. preferably autoimmune diseases, circulatory diseases, tumor diseases, immunodeficiency diseases, diseases that affect individual organs or entire organ areas or tissues, as well as diseases that are caused by viruses, bacteria or parasites.
  • the invention also relates to a method for loading the active substance into the liposomes, a defined pH value being used for encapsulation and a second pH value being set for separating the unbound active substance.
  • the invention also relates to the use of the liposomes for the production of nanocapsules.
  • the invention further relates to the use of the liposomes for the production of release systems in diagnostics.
  • the liposomes are advantageously used to transport and / or release active substances.
  • the liposomes can advantageously be used in particular in the case of intravenous or peritoneal administration.
  • the use of the liposomes as a vector for the transfection of cells in vivo, in vitro and ex vivo is advantageous.
  • Cargomolecules that are not installed and adhere to the outside can, if necessary, be increased by simply increasing the pH be removed. This step is always necessary if non-built-in cargo molecules lead to an aggregation of the liposomes.
  • An advantage of using the components according to the invention is the fact that the enclosed active substances only have to be brought under conditions that permit interaction with the lipid layer for the period of the actual inclusion. As soon as the lipid layer remains closed, you can switch to other conditions. A possible inactivation of active substances, especially proteins, can be minimized.
  • Liposomes comprising the components according to the invention can be coated with polymers under conditions known to those skilled in the art.
  • single or multiple deposition of such substances can be carried out on the surface.
  • a multiple deposition which may be carried out in the presence of a crosslinking agent, results in liposomal nanocapsules as described in WO 00/28972 or in WO01 / 64330 and included in the disclosure content of the invention.
  • An advantage of using the substances described here is the fact that the electrostatic interaction with the polyelectrolyte can be interrupted. It is known that the interaction of a polyelectrolyte with charge carriers of the liposomal membrane can lead to segregation of membrane components and to the formation of lipid clusters. In many cases, this segregation is accompanied by permeabilization of the liposome.
  • the substances according to the invention enable this interaction to be switched off after the coating process. Will the pH at this point increased, the liposomes are only sterically enclosed in the nanocapsules, and the membrane no longer interacts with the polyelectrolytes. Clustering of the lipids and the associated permeabilization of the membrane can thus be avoided.
  • the changes in permeability are used specifically for loading liposomes.
  • An active substance to be enclosed can be added to the medium under conditions of high permeability. Conditions of low permeability are then set. The active ingredient thus remains inside the liposomes. Active substance not included can then optionally be separated off.
  • Such a change in permeability can be brought about on liposomes or on liposomal nanocapsules.
  • the liposomes according to the invention easily fuse with other membranes at low pH. Usually this step requires the presence of a greater proportion of PE in the membrane. Due to its tendency to form hexgonal phases, this has the function of a helper lipid. However, the lower stability of such membranes is disadvantageous, and a gradual release of trapped active substances is often observed here.
  • liposomes which are produced using the substances according to the invention effectively fuse even in the absence of such a helper lipid.
  • Using the substances according to the invention it is therefore possible to produce those liposomes which can stably encapsulate an active ingredient, but fuse under the conditions of a low pH with cell membranes and release the active ingredient there.
  • An essential prerequisite for the use of liposomes for experimental or therapeutic purposes is their compatibility with cells and tissues.
  • a number of known compounds which are used for the introduction of DNA or proteins into cells are cytotoxic.
  • the compounds according to the invention show a reduced cytotoxicity. This is particularly the case when the compounds contain physiological building blocks such as amino acids.
  • vectors for gene or protein transport in cells are compatibility with serum or blood. Because of their strong cationic charge, the currently known vectors form uncontrolled large aggregates with serum, which lead to the formation of thrombi in the organism. Their use is practically excluded in vivo and limited to in vitro or ex vivo applications.
  • liposomes which have been built up using the components according to the invention do not form aggregates in serum or blood.
  • liposomes are attacked by the components of the complement system and quickly lysed. This reaction takes place within minutes. Pore formation occurs in the membrane, through which even large ones Diffuse out molecules like proteins. Stabilization of liposomes against these mechanisms has so far only been possible by incorporating cholesterol into the lipid layer. Such liposomes are then very stable, but can no longer interact with cells or release their active ingredient easily. It has surprisingly been found that liposomes which are built up using the components according to the invention are stable in serum or blood for several hours. The drug release is low even under these conditions.
  • a liposomal vector for the transport of active substances must meet at least three requirements: it must have low toxicity, contain the active substance safely and stably and be compatible with serum or blood.
  • liposomes that are produced using the substances according to the invention.
  • the liposomes disclosed here are therefore well suited for use in therapeutic purposes.
  • Other properties that support this use are the good loadability with active substances and the targeted release of these substances by permeabilization of the membrane at suitable pH values.
  • dipalmitoylglycerol succinate 1.7 g dipalmitoylglycerol succinate are dissolved in 20 ml DMF at room temperature. 400 mg of carbonyldiimidazole, dissolved in 20 ml of DMF, are added to the solution. The mixture is allowed to stir for 1 hour and then 300 mg of histamine are added. After the mixture was stirred overnight, the mixture was concentrated completely in vacuo. The residue is purified by column chromatography on silica gel 60; chloroform / methanol 10: 1 is used as the eluent.
  • Buffer (10 mM KAc, 10 mM HEPES, 150 mM NaCl, pH7.5) in a lipid concentration of 5mM hydrated by short ultrasound exposure (5 min). Finally, the emulsion is frozen and extruded several times after thawing (Avestin LiposoFa ⁇ t, polycarbonate filter 200nm pore size). The course of the zeta potential at different pH values is shown in the table below.
  • Unilamellar liposomes (DPPC / DPPG / cholesterol 40:20:40 mol%) are suspended in a concentration of 20 mM lipid in a borate buffer (20 mM sodium borate, 120 mM sodium chloride pH 8.4). 400 ⁇ l of one are added to 2 ml of this solution
  • the eluate of the liposome suspension is made up to 4 ml.
  • Histamine is added to the oxidized liposomes in a final concentration of 20 mM and incubated for 2 hours. Finally, the mixture was reduced with 20 mM sodium borohydride overnight at 4 ° C. Excess histamine can be separated by chromatography on Sephadex G25 as above.
  • lipid film Depending on the lipid composition of the lipid film, 1.43 mM of the pH-sensitive lipid are dissolved in chloroform with the other lipids. After the solvent has been stripped off, the lipid film is dried in vacuo overnight.
  • the lipid film is hydrated directly with 1 ml of DNA-containing (100 ⁇ g DNA / ml) NaAc buffer (10 mM NaAc, 150 mM NaCl, pH 4) (light ultrasound, then rotated for 30 min above the phase transition temperature). Then there is a freeze / thaw step.
  • DNA that is not included can be separated by flotation in a sucrose gradient (at pH 7.5) (0.8 M sucrose, 0.5 M sucrose, buffer).
  • the DNA content is determined with the help of the intercalation dye propidium iodide through the increase in the fluorescence intensity when intercalated into the DNA.
  • the intercalation dye propidium iodide For this purpose, 20 ⁇ l propidium iodide, 6 ⁇ l Triton X-100 (10% in water) are filled with sample to 300 ⁇ l and measured with a fluorescence plate reader.

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Families Citing this family (45)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003093449A2 (en) * 2002-05-06 2003-11-13 Nucleonics, Inc. Methods for delivery of nucleic acids
EP1706149A2 (en) * 2004-01-15 2006-10-04 Alza Corporation Liposome composition for delivery of therapeutic agents
KR20080082956A (ko) 2005-09-15 2008-09-12 노보솜 아게 양쪽성 리포솜의 개선 또는 양쪽성 리포솜에 관련된 개선
US20080088046A1 (en) * 2006-10-13 2008-04-17 Steffen Panzner Amphoteric liposomes, a method of formulating an amphoteric liposome and a method of loading an amphoteric liposome
US8101741B2 (en) 2005-11-02 2012-01-24 Protiva Biotherapeutics, Inc. Modified siRNA molecules and uses thereof
CA2631677C (en) * 2005-12-01 2014-08-12 Pronai Therapeutics, Inc. Amphoteric liposome formulation
RU2451022C2 (ru) * 2005-12-15 2012-05-20 Сантр Насьональ Де Ля Решерш Сьентифик (Снрс) Катионные олигонуклеотиды, автоматизированные способы их получения и их применение
EP1844772A1 (en) * 2006-04-06 2007-10-17 Polyplus-Transfection SA Compositions for transfection of oligonucleotides active for gene silencing and their biological and therapeutical applications
JP5424885B2 (ja) 2006-10-13 2014-02-26 マリーナ バイオテック，インコーポレイテッド 両性リポソームにおけるまたは両性リポソームに関する改善、両性リポソームを処方する方法および両性リポソームに充填する方法
WO2008074487A2 (en) * 2006-12-19 2008-06-26 Novosom Ag Lipids and lipid assemblies comprising transfection enhancer elements
WO2009082817A1 (en) 2007-12-27 2009-07-09 Protiva Biotherapeutics, Inc. Silencing of polo-like kinase expression using interfering rna
FR2926818B1 (fr) * 2008-01-30 2012-04-06 Centre Nat Rech Scient siRNA CATIONIQUES, SYNTHESE ET UTILISATION POUR L'ARN INTERFERENCE
JP2011516094A (ja) 2008-04-15 2011-05-26 プロチバ バイオセラピューティクス インコーポレイティッド 干渉ｒｎａを用いたｃｓｎ５遺伝子発現のサイレンシング方法
JP5588619B2 (ja) * 2009-03-11 2014-09-10 一丸ファルコス株式会社 ｐＨ応答性リポソーム
CA2784568A1 (en) 2009-12-18 2011-06-23 The University Of British Columbia Lipid particles for delivery of nucleic acids
EP2582387A2 (en) 2010-06-17 2013-04-24 The United States Of America As Represented By The Secretary, National Institutes Of Health Compositions and methods for treating inflammatory conditions
PL2591114T3 (pl) 2010-07-06 2017-08-31 Glaxosmithkline Biologicals Sa Immunizacja dużych ssaków małymi dawkami rna
HUE047796T2 (hu) 2010-07-06 2020-05-28 Glaxosmithkline Biologicals Sa RNS bevitele több immunútvonal bekapcsolására
RS63817B1 (sr) 2010-07-06 2023-01-31 Glaxosmithkline Biologicals Sa Čestice za isporuku slične virionu za molekule samoreplicirajuće rnk
HUE026646T2 (en) 2010-07-06 2016-07-28 Glaxosmithkline Biologicals Sa Preferred liposomes containing lipids of PKA value for delivery of RNA
PT3970742T (pt) 2010-08-31 2022-06-27 Glaxosmithkline Biologicals Sa Lipossomas peguilados para entrega de arn codificador de imunogénio
JP2013544504A (ja) 2010-10-11 2013-12-19 ノバルティス アーゲー 抗原送達プラットフォーム
US10849975B2 (en) 2011-02-03 2020-12-01 Thomas Jefferson University Multivalent vaccines for rabies virus and filoviruses
WO2012109495A1 (en) 2011-02-09 2012-08-16 Metabolic Solutions Development Company, Llc Cellular targets of thiazolidinediones
US8789967B2 (en) 2011-06-02 2014-07-29 Musco Corporation Apparatus, method, and system for independent aiming and cutoff steps in illuminating a target area
WO2013006838A1 (en) 2011-07-06 2013-01-10 Novartis Ag Immunogenic combination compositions and uses thereof
WO2013032643A2 (en) * 2011-08-31 2013-03-07 Dicerna Pharmaceuticals, Inc. Lipids capable of conformational change and their use in formulations to deliver therapeutic agents to cells
AU2012340086A1 (en) 2011-11-17 2014-05-29 The Regents Of The University Of California Therapeutic RNA switches compositions and methods of use
US9035039B2 (en) 2011-12-22 2015-05-19 Protiva Biotherapeutics, Inc. Compositions and methods for silencing SMAD4
CN104884065B (zh) 2012-09-21 2019-01-01 强烈治疗剂公司 治疗癌症的方法
BR112015010220A2 (pt) 2012-11-05 2017-12-05 Pronai Therapeutics Inc métodos de utilização de biomarcadores para tratamento de câncer
JP6339884B2 (ja) 2014-07-17 2018-06-06 富士フイルム株式会社 イミダゾール化合物およびそれを含有するリポソーム
BR112017006679A2 (pt) 2014-10-02 2017-12-26 Protiva Biotherapeutics Inc moléculas, composição, partícula, composição farmacêutica, métodos para silenciar a expressão de um gene, usos de uma partícula, métodos para melhorar um ou mais sintomas, métodos para tratar uma infecção, usos de uma composição, método para inibir a replicação do vírus da hepatite d
US20180245074A1 (en) 2015-06-04 2018-08-30 Protiva Biotherapeutics, Inc. Treating hepatitis b virus infection using crispr
CN108350455A (zh) 2015-07-29 2018-07-31 阿布特斯生物制药公司 用于使b型肝炎病毒基因表达沉默的组合物和方法
WO2017040702A1 (en) * 2015-08-31 2017-03-09 The Regents Of The University Of California Bipolar tetraether lipids
MX2018010198A (es) 2016-02-25 2019-06-12 Applied Biological Laboratories Inc Composiciones y metodos para proteger contra patogenos e irritantes transportados por el aire.
US12447166B2 (en) 2016-02-25 2025-10-21 Applied Biological Laboratories, Inc. Compositions and methods for protecting against airborne pathogens and irritants
WO2017176596A1 (en) 2016-04-04 2017-10-12 The United States Of America, As Represented By The Secretary, Department Of Health & Human Services Multivalent vaccines for rabies virus and coronaviruses
EP3500294A4 (en) 2016-08-22 2020-07-29 Arbutus Biopharma Corporation ANTI-PD-1 ANTIBODIES, OR THEIR FRAGMENTS, FOR THE TREATMENT OF HEPATITIS B
US11198831B2 (en) 2019-01-31 2021-12-14 Kvi Llc Lubricant for a device
KR20230011913A (ko) 2020-03-04 2023-01-25 버브 테라퓨틱스, 인크. 표적화된 rna 전달을 위한 조성물 및 방법
JP7848179B2 (ja) * 2021-03-09 2026-04-20 日本精化株式会社 リン脂質
JP2024530647A (ja) 2021-08-03 2024-08-23 ヴァーヴ・セラピューティクス，インコーポレーテッド 標的ｒｎａ送達のための組成物および方法
CN115894281B (zh) * 2021-09-22 2025-07-01 广州谷森制药有限公司 阳离子脂质化合物、其制备方法、组合物及应用

Family Cites Families (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE45744T1 (de) * 1984-07-25 1989-09-15 Ciba Geigy Ag Phosphatidylverbindungen, verfahren zu ihrer herstellung und ihre verwendung.
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
US5264618A (en) * 1990-04-19 1993-11-23 Vical, Inc. Cationic lipids for intracellular delivery of biologically active molecules
US5334761A (en) * 1992-08-28 1994-08-02 Life Technologies, Inc. Cationic lipids
FR2701396B1 (fr) * 1993-02-12 1995-04-21 Oreal Procédé de stabilisation de vésicules de lipide(s) amphiphile(s) et composition pour application topique contenant lesdites vésicules stabilisées.
US6020317A (en) * 1993-02-19 2000-02-01 Nippon Shinyaku Co. Ltd. Glycerol derivative, device and pharmaceutical composition
US5354853A (en) * 1993-03-12 1994-10-11 Genzyme Corporation Phospholipid-saccharide conjugates
US5965434A (en) * 1994-12-29 1999-10-12 Wolff; Jon A. Amphipathic PH sensitive compounds and delivery systems for delivering biologically active compounds
FR2756565B1 (fr) * 1996-12-04 1998-12-31 Oreal Nouveaux derives d'histidine et leur utilisation en cosmetique ou en pharmacie
JPH11199488A (ja) * 1998-01-09 1999-07-27 Sankyo Co Ltd 抗腫瘍薬のリポソーム化製剤
JPH11217328A (ja) * 1998-01-28 1999-08-10 Nissin Food Prod Co Ltd 抗ｈｉｖ活性を発現する脂質集合体
US6656498B1 (en) 1998-11-25 2003-12-02 Vanderbilt University Cationic liposomes for gene transfer
US7112337B2 (en) * 1999-04-23 2006-09-26 Alza Corporation Liposome composition for delivery of nucleic acid
WO2001026625A2 (en) * 1999-10-08 2001-04-19 Alza Corp Neutral-cationic lipid for nucleic acid and drug delivery
DE10109898A1 (de) * 2001-02-21 2002-09-05 Novosom Gmbh Lipide mit veränderlicher Ladung
DE10109897A1 (de) * 2001-02-21 2002-11-07 Novosom Ag Fakultativ kationische Liposomen und Verwendung dieser

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO03070220A1 *

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