WO2009129395A1 - Lipides cationiques et utilisations de ceux-ci - Google Patents

Lipides cationiques et utilisations de ceux-ci Download PDF

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WO2009129395A1
WO2009129395A1 PCT/US2009/040817 US2009040817W WO2009129395A1 WO 2009129395 A1 WO2009129395 A1 WO 2009129395A1 US 2009040817 W US2009040817 W US 2009040817W WO 2009129395 A1 WO2009129395 A1 WO 2009129395A1
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weight
lipid
particle
octadeca
bis
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PCT/US2009/040817
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Prasad A. Dande
Todd M. Hansen
Robert D. Hubbard
Carol K. Wada
Lu Tian
Xiaobin Zhao
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Abbott Laboratories
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    • 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
    • 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
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    • C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D207/12—Oxygen or sulfur atoms
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    • C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D207/14—Nitrogen atoms not forming part of a nitro radical
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    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D213/28—Radicals substituted by singly-bound oxygen or sulphur atoms
    • C07D213/30—Oxygen atoms
    • C—CHEMISTRY; METALLURGY
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    • 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
    • C07D233/56—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 only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
    • C07D233/60—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 only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms with hydrocarbon radicals, substituted by oxygen or sulfur atoms, attached to ring nitrogen atoms
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    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/20—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D239/22—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with hetero atoms directly attached to ring carbon atoms
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    • C07D243/00—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms
    • C07D243/06—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4
    • C07D243/08—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4 not condensed with other rings
    • 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
    • C—CHEMISTRY; METALLURGY
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D317/00—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms
    • C07D317/08—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3
    • C07D317/10—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3 not condensed with other rings
    • C07D317/14—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3 not condensed with other rings with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D317/18—Radicals substituted by singly bound oxygen or sulfur atoms
    • C07D317/24—Radicals substituted by singly bound oxygen or sulfur atoms esterified
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
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    • C07D317/00—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms
    • C07D317/08—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3
    • C07D317/10—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3 not condensed with other rings
    • C07D317/14—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3 not condensed with other rings with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D317/28—Radicals substituted by nitrogen atoms
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/06—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms

Definitions

  • This invention pertains to cationic lipids, cationic lipid based drug delivery systems, ways to make them, and methods of treating diseases using them.
  • RNA Ribonucleic acid
  • RNA Ribonucleic acid
  • antisense oligonucleotide a DNA, a plasmid, a ribosomal RNA (rRNA), a micro RNA (miRNA), transfer RNA (tRNA), a small inhibitory RNA (siRNA), and small nuclear RNA (snRNA).
  • rRNA ribosomal RNA
  • miRNA micro RNA
  • tRNA transfer RNA
  • siRNA small inhibitory RNA
  • snRNA small nuclear RNA
  • Such novel delivery formulations will need, for example, to allow for appropriate internalization of the therapeutic agent into the cell, agents sufficient absorption from the site of administration, distribution to various tissues, sufficient residence time and concentration at the sites of action to elicit effective biologic response, in addition to also maintaining it's stability, and size.
  • PEG polyethylene glycol
  • One embodiment of this invention therefore pertains to a cationic lipid or mixtures thereof, having Formula (I)
  • R 1 and R2 are independently H, cycloalkyl, cycloalkenyl or R 5 ; or
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl or heteroaryl;
  • L 1 is O, OC(O) or (O)CO
  • L 2 is O, OC(O) or (O)CO
  • R 3 and R 4 is H, and the other is C 14 -C 20 -alkenyl or C 14 -C 20 -alkyl; or
  • R and R are independently C 14 -C 20 -alkenyl or C 14 -C 20 -alkyl; or
  • R and R combine to form CR R , wherein R is H and R is C 14 -C 20 -alkenyl or C 14 -C 20 -alkyl; or R and R are the same or are different and are C 14 -C 20 -alkenyl, C 14 -C 20 - alkyl, or (CH 2 O)-C 14 -C 20 alkenyl;
  • R is alkyl, which is unsubstituted or substituted with one or more R , OR , SR , S(O)R 6 , SO 2 R 6 , C(O)R 6 , CO(O)R 6 , OC(O)R 6 , OC(O)OR 6 , NH 2 , NHR 6 , N(R 6 ) 2 , NHC(O)R 6 , NR 6 C(O)R 6 , NHS(O) 2 R 6 , N R 6 S(O) 2 R 6 , NHC(O)OR 6 , NR 6 C(O)OR 6 , NHC(O)NH 2 , NHC(O)NHR 6 , NHC(O)N(R 6 ) 2 , NR 6 C(O)NHR 6 , NR 6 C(O)N(R 6 ) 2 , C(O)NH 2 , C(O)NHR 6 , C(O)N(R 6 ) 2 , C(O)
  • R 6 is R 7 , R 8 , R 9 , or R 10 ;
  • R is phenyl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R 8 is heteroaryl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R 10 is alkyl, alkenyl or alkynyl
  • each foregoing cyclic moiety is independently unsubstituted or substituted with one or more R 11 , OR 11 , SR 11 , S(O)R 11 , SO 2 R 11 , C(O)R 11 , CO(O)R 11 , OC(O)R 11 , OC(O)OR 11 , NH 2 , NHR 11 , N(R 1 1 ) 2 , NHC(O)R 11 , NR 11 C(O)R 11 , NHS(O) 2 R 11 , N R 11 S(O) 2 R 11 , NHC(O)OR 11 , NR 11 C(O)OR 11 , NHC(O)NH 2 , NHC(O)NHR 11 , NHC(O)N(R 1 1 ) 2 , NR 11 C(O)NHR 11 , NR 11 C(O)N(R 1 1 ) 2 , C(O)NH 2 , C(O)NHR 11 ,
  • R 11 is R 12 , R 13 , R 14 , or R 15 ;
  • R is phenyl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R 13 is heteroaryl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R 14 is cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, each of which is unfused or fuse
  • R is alkyl, alkenyl, alkynyl, or R ;
  • R is phenyl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R , R , R , and R are independently unsubstituted or substituted with one or more R 18 , OR 18 , SR 18 , S(O)R 18 , SO 2 R 18 , C(O)R 18 , CO(O)R 18 , OC(O)R 18 , OC(O)OR 18 , NH 2 , NHR 18 , N(R 18 ) 2 , NHC(O)R 18 , NR 18 C(O)R 18 , NHS(O) 2 R 18 , NR 18 S(O) 2 R 18 , NHC(O)OR 18 , NR 18 C(O)OR 18 , NHC(O)NH 2 , NHC(O)NHR 18 , NHC(O)N(R 18 ) 2 , NR 18 C(O)NHR 18 , NR 18 C(O)N(R 18 ) 2 , C(O)NH 2 , C(O)NHR 18 , C(O)N(R 18
  • a further embodiment pertains to Cationic-Based Lipid Encapsulation Systems (CaBLES) comprising one or more non-cationic lipids, one or more polyethylene glycol (PEG)-lipid conjugates and one or more cationic lipids having Formula I.
  • CaBLES Cationic-Based Lipid Encapsulation Systems
  • cationic lipids of the present invention i.e., cationic lipids of Formula I
  • cationic lipids of Formula I can be used in the preparation of either empty liposomes or used to deliver any product (e.g., therapeutic agents including nucleic acids, diagnostic agents, labels or other compounds) to a cell tissue, including cells and tissues in mammals.
  • Lipid-Based Particles of the present invention are defined as CaBLES which further comprise one or more therapeutic agent(s).
  • Such Lipid- Based Particles can be used to deliver any of a variety of therapeutic agent(s), preferably said therapeutic agent is a nucleic acid encoded with a product of interest, including but not limited to, RNA, antisense oligonucleotide, a DNA, a plasmid, a ribosomal RNA (rRNA), a micro RNA (miRNA), transfer RNA (tRNA), a small inhibitory RNA (siRNA), small nuclear RNA (snRNA), antigens, fragments thereof, proteins, peptides, vaccines and small-molecules or mixtures thereof.
  • RNA nucleic acid encoded with a product of interest
  • rRNA ribosomal RNA
  • miRNA micro RNA
  • tRNA transfer RNA
  • siRNA small inhibitory RNA
  • snRNA small nuclear RNA
  • a further embodiment pertains to pharmaceutical compositions comprising a Lipid- Based Particle and a pharmaceutically acceptable carrier.
  • a further embodiment pertains to a method of treating cancer in a mammal comprising administering thereto a therapeutically acceptable amount of a Lipid-Based Particle.
  • Yet another embodiment pertains to a method of decreasing tumor volume in a mammal comprising administering thereto a therapeutically acceptable amount of a Lipid- Based Particle.
  • a further embodiment pertains to a method of making CaBLES or Lipid-Based Particles, comprising: (a) mixing the cationic lipid(s), the non-cationic lipid(s) and the PEG- lipid conjugate(s); (b) adding the mixture of step (a) to one or more therapeutic agents; and (c) separating and purifying resulting suspension of step (b).
  • This invention pertains to in vitro and in vivo delivery of therapeutic agents.
  • the invention pertains to compositions that allow for delivery of nucleic acids, including but not limited to RNA, antisense oligonucleotide, a DNA, a plasmid, a ribosomal RNA (rRNA), a micro RNA (miRNA), transfer RNA (tRNA), a small inhibitory RNA (siRNA), small nuclear RNA(snRNA), antigens, fragments thereof, proteins, peptides, and small molecules.
  • nucleic acids including but not limited to RNA, antisense oligonucleotide, a DNA, a plasmid, a ribosomal RNA (rRNA), a micro RNA (miRNA), transfer RNA (tRNA), a small inhibitory RNA (siRNA), small nuclear RNA(snRNA), antigens, fragments thereof, proteins, peptides, and small molecules.
  • Variable moieties of compounds herein are represented by identifiers (capital letters with numerical and/or alphabetical superscripts) and may be specifically embodied.
  • variable moiety may be the same or different as another specific embodiment having the same identifier and that asymmetric divalent moieties are drawn from left to right.
  • alkenyl means monovalent, straight or branched chain hydrocarbon moieties having one or more than one carbon-carbon double bonds, such as C 2 - alkenyl, C 3 -alkenyl, C 4 -alkenyl, Cs-alkenyl, C ⁇ -alkenyl and the like.
  • C 1 -C ⁇ -alkylene means divalent, saturated, straight or branched chain hydrocarbon moieties bonds, such as C 1 -alkylene, C2-alkylene, C3-alkylene, C4-alkylene, Cs-alkylene and C ⁇ -alkylene.
  • alkyl means monovalent, straight or branched chain hydrocarbon moieties such as C 1 -alkyl, C 2 -alkyl, C 3 -alkyl, C 4 -alkyl, Cs-alkyl and C ⁇ -alkyl.
  • alkynyl means monovalent, straight or branched chain hydrocarbon moieties having one or more than one carbon-carbon triple bonds, such as C 2 -alkynyl, C 3 -alkynyl, C 4 -alkynyl, Cs-alkynyl, C ⁇ -alkynyl and the like.
  • C 1 -Cg-alkyl as used herein, means C 1 -alkyl, C 2 -alkyl, C 3 -alkyl, C 4 -alkyl, Cs-alkyl, C 6 -alkyl, C 7 -alkyl and C 8 -alkyl.
  • C 14 -C 20 -alkenyl means C 1 4-alkenyl,” C 15 - alkenyl,” C 16 - alkenyl,” C n - alkenyl,” C 18 - alkenyl,” C 19 - alkenyl” and C 20 - alkenyl.”
  • C 14 -C 20 -alkyl means C 14 -alkyl,” C 15 -alkyl,” C ⁇ -alkyl,” C ⁇ -alkyl,” C 18 -alkyl,” C 19 -alkyl” and C 20 -alkyl.”
  • cycloalkane means saturated cyclic or bicyclic hydrocarbon moieties, such as C3-cycloalkane, C4-cycloalkane, Cs-cycloalkane, C ⁇ -cycloalkane and the like.
  • cycloalkyl means monovalent, saturated cyclic and bicyclic hydrocarbon moieties, such as C 3 -cycloalkyl, C 4 -cycloalkyl, C 5 -cycloalkyl, C ⁇ -cycloalkyl and the like.
  • cycloalkene means cyclic and bicyclic hydrocarbon moieties having one or more than one carbon-carbon double bonds, such as Cs-cycloalkene, C ⁇ -cycloalkene and the like.
  • cycloalkenyl means monovalent, cyclic hydrocarbon moieties having one or more than one carbon-carbon double bonds, such as C 4 -cycloalkenyl, C 5 -cycloalkenyl, C ⁇ -cycloalkenyl and the like.
  • heteroene means a f ⁇ ve-membered or six-membered aromatic ring having at least one carbon atom and one or more than one independently selected nitrogen, oxygen or sulfur atom.
  • the heteroarenes of this invention are connected through any adjacent atoms in the ring, provided that proper valences are maintained.
  • heteroarenes include, but are not limited to furan, imidazole, isothiazole, isoxazole, oxadiazole, oxazole, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, thiazole, thiadiazole thiophene, tetrazine, tetrazole, triazine, triazole and the like.
  • heteroaryl means a monovalent f ⁇ ve-membered or six- membered aromatic ring having at least one carbon atom and one or more than one independently selected nitrogen, oxygen or sulfur atom.
  • the heteroaryls of this invention are connected through any carbon atom or any nitrogen atom in the ring, provided that proper valences are maintained.
  • heteroaryls include, but are not limited to, furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazinyl, triazolyl and the like.
  • heterocycloalkane means cycloalkane having one or two or three CH 2 moieties replaced with independently selected O, S, S(O), SO 2 or NH and one or two CH moieties unreplaced or replaced with N and also means cycloalkane having one or two or three CH 2 moieties unreplaced or replaced with independently selected O, S, S(O), SO 2 or NH and one or two CH moieties replaced with N.
  • heterocycloalkene means cycloalkene having one or two or three CH 2 moieties replaced with independently selected O, S, S(O), SO 2 or NH and one or two CH moieties unreplaced or replaced with N and also means cycloalkene having one or two or three CH 2 moieties unreplaced or replaced with independently selected O, S, S(O), SO 2 or NH and one or two CH moieties replaced with N.
  • heterocycloalkyl means cycloalkyl having one or two or three CH 2 moieties replaced with independently selected O, S, S(O), SO 2 or NH and one or two CH moieties unreplaced or replaced with N and also means cycloalkyl having one or two or three CH 2 moieties unreplaced or replaced with independently selected O, S, S(O), SO 2 or NH and one or two CH moieties replaced with N.
  • heterocycloalkenyl means cycloalkenyl having one or two or three CH 2 moieties replaced with independently selected O, S, S(O), SO 2 or NH and one or two CH moieties unreplaced or replaced with N and also means cycloalkenyl having one or two or three CH 2 moieties unreplaced or replaced with independently selected O, S, S(O), SO 2 or NH and one or two CH moieties replaced with N.
  • cyclic moiety means benzene, cycloalkane, cycloalkyl, cycloalkene, cycloalkenyl, heteroarene, heteroaryl, heterocycloalkane, heterocycloalkyl, heterocycloalkene, heterocycloalkenyl and phenyl.
  • DSPC 1,2-distearoyl-5/?-glycero-3-phosphocholine.
  • Choi as used herein, means cholesterol.
  • PEG-Chol means poly(oxy-1,2-ethanediyl)-2000- ⁇ -(3 ⁇ )- cholest-5-en-3-yl-omega-hydroxy.
  • Pal-PEG-Cera N-palmitoyl-sphingosine-1- [succinyl(methoxypolyethylene glycol)-2000] .
  • PEG-DMPE N-(carbonyl- methoxypolyethyleneglycol-2000)-1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine.
  • PEG-DPPE N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine.
  • PEG-DSPE N-(carbonyl- methoxypolyethyleneglycol-2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine.
  • PEG-DMG means 1 ,2-dimyristoyl-sn-glycerol- methoxypolyethyleneglycol-2000.
  • PEG-DPG means 1 ,2-dipalmitoyl-sn-glycerol- methoxypolyethyleneglycol-2000.
  • PEG-DSG means 1 ,2-distearoyl-sn-glycerol- methoxypolyethyleneglycol-2000.
  • MALDI matrix assisted laser desorption ionization
  • particle means a small object that behaves as a whole unit in terms of its transport and properties.
  • nanoparticle means any particle having a diameter of less than 1000 nanometers. In some embodiments, nanoparticles have a diameter of 500 or less. In some embodiments, nanoparticles have a diameter of 200 or less.
  • nucleic acid or “polynucleotide” refers to a polymer containing at least two deoxyribonucleotides or ribonucleotides in either single- or double-stranded form.
  • Nucleic acids include nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides.
  • analogs include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs).
  • PNAs peptide-nucleic acids
  • the terms encompasses nucleic acids containing known analogues of natural nucleotides that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides.
  • a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions), alleles, orthologs, SNPs, and complementary sequences as well as the sequence explicitly indicated.
  • degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or deoxyinosine residues (Batzer et al., Nucleic Acid Res. 19:5081 (1991); Ohtsuka et al., J. Biol. Chem. 260:2605-2608 (1985); and Cassol et al. (1992); Rossolini et al., MoI. Cell. Probes 8:91-98 (1994)).
  • "Nucleotides” contain a sugar deoxyribose (DNA) or ribose (RNA), a base, and a phosphate group.
  • Nucleotides are linked together through the phosphate groups. Nucleotides include chemically modified nucleotides as described in, e.g., WO 03/74654. "Bases" include purines and pyrimidines, which further include natural compounds adenine, thymine, guanine, cytosine, uracil, inosine, and natural analogs, and synthetic derivatives of purines and pyrimidines, which include, but are not limited to, modifications which place new reactive groups such as, but not limited to, amines, alcohols, thiols, carboxylates, and alkylhalides.
  • DNA may be in the form of antisense, plasmid DNA, parts of a plasmid DNA, pre-condensed DNA, product of a polymerase chain reaction (PCR), vectors (Pl, PAC, BAC, YAC, artificial chromosomes), expression cassettes, chimeric sequences, chromosomal DNA, or derivatives of these groups.
  • PCR polymerase chain reaction
  • vectors Pl, PAC, BAC, YAC, artificial chromosomes
  • expression cassettes chimeric sequences, chromosomal DNA, or derivatives of these groups.
  • nucleic acid is used interchangeably with gene, plasmid, cDNA, mRNA, and an interfering RNA molecule (e.g. a synthesized siRNA or an siRNA expressed from a plasmid).
  • RNA means a small inhibitory RNA, and molecules having endogenous RNA bases or chemically modified nucleotides. The modifications shall not abolish cellular activity, but rather impart increased stability and/or increased cellular potency. Examples of chemical modifications include phosphorothioate groups, T- deoxynucleotide, 2'-OCH3-containing ribonucleotides, 2'-F-ribonucleotides, 2'-methoxyethyl ribonucleotides or a combination thereof.
  • SPC soybean phosphatidylcholine
  • small molecule means antibiotics, antineoplastics, antiinflammatories, anitivirals, immunomodulators and agents that act upon the respiratory system, the cardiovascular system, the central nervous system or a metabolic pathway involved with dyslipidemia, diabetes or Syndrome X.
  • Compounds of this invention may contain asymmetrically substituted carbon atoms in the R or S configuration, wherein the terms "R” and “S” are as defined in Pure Appl. Chem. (1976) 45, 13-10.
  • Compounds having asymmetrically substituted carbon atoms with equal amounts of R and S configurations are racemic at those atoms. Atoms having excess of one configuration over the other are assigned the configuration in excess, preferably an excess of about 85%-90%, more preferably an excess of about 95%-99%, and still more preferably an excess greater than about 99%. Accordingly, this invention is meant to embrace racemic mixtures and relative and absolute diastereoisomers and the compounds thereof.
  • Compounds of this invention may also contain carbon-carbon double bonds or carbon-nitrogen double bonds in the E or Z configuration, wherein the term “E” represents higher order substituents on opposite sides of the carbon-carbon or carbon-nitrogen double bond and the term “Z” represents higher order substituents on the same side of the carbon- carbon or carbon-nitrogen double bond as determined by the Cahn-Ingold-Prelog Priority Rules.
  • the compounds of this invention may also exist as a mixture of "E” and "Z" isomers.
  • One embodiment of this invention therefore pertains to a cationic lipid or mixtures thereof, having Formula (I)
  • R and R are independently H, cycloalkyl, cycloalkenyl or R ; or
  • L 1 is O, OC(O) or (O)CO
  • L 2 is O, OC(O) or (O)CO
  • R and R is H, and the other is C 14 -C 20 -alkenyl or C 14 -C 20 -alkyl; or
  • R and R are independently C 14 -C 20 -alkenyl or C 14 -C 20 -alkyl; or 3 4 20 21 20 21
  • R and R combine to form CR R , wherein R is H and R is C 14 -C 20 -alkenyl or
  • C 14 -C 20 -alkyl or R and R are the same or are different and are C 14 -C 20 -alkenyl, C 14 -C 20 - alkyl, or (CH 2 O)-C 14 -C 2 O alkenyl;
  • R is alkyl, which is unsubstituted or substituted with one or more R , OR , SR ,
  • r R> 6 i • s r R> 7 , r R> 8 , r R> 9 , or r R> 10 ;
  • R is phenyl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is heteroaryl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is alkyl, alkenyl or alkynyl
  • each foregoing cyclic moiety is independently unsubstituted or substituted with one or more R 11 , OR 11 , SR 11 , S(O)R 11 , SO 2 R 11 , C(O)R 11 , CO(O)R 11 , OC(O)R 11 , OC(O)OR 11 , NH 2 , NHR 11 , N(R 1 ⁇ 2 , NHC(O)R 11 , NR 11 C(O)R 11 , NHS(O) 2 R 11 , N R 1 ! S(O) 2 R 1 ! , NHC(O)OR 1 ⁇ NR U C(O)OR 1 ! , NHC(O)NH 2 , NHC(O)NHR 1 !
  • R is R , R , R , or R ;
  • R is phenyl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is heteroaryl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is alkyl, alkenyl or alkynyl, each of which is unsubstituted or substituted with one or two of independently selected R 16 , OR 16 , SR 16 , S(O) 2 R 16 , C(O)OH, NH 2 , NHR 16 N(R 16 ) 2 , C(O)R 16 , C(O)NH 2 , C(O)NHR 16 , C(O)N(R 16 ) 2 , NHC(O)R 16 , NR 16 C(O)R 16 , NHC(O)OR 16 , NR 16 C(O)OR 16 , OH, F, Cl, Br or I;
  • R is alkyl, alkenyl, alkynyl, or R ;
  • R is phenyl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R , R , R , and R are independently unsubstituted or substituted with one or more R 18 , OR 18 , SR 18 , S(O)R 18 , SO 2 R 18 , C(O)R 18 , CO(O)R 18 , OC(O)R 18 , OC(O)OR 18 , NH 2 , NHR 18 , N(R 18 ) 2 , NHC(O)R 18 , NR 18 C(O)R 18 , NHS(O) 2 R 18 , NR 18 S(O) 2 R 18 , NHC(O)OR 18 , NR 18 C(O)OR 18 , NHC(O)NH 2 , NHC(O)NHR 18 , NHC(O)N(R 18 ) 2 , NR 18 C(O)NHR 18 , NR 18 C(O)N(R 18 ) 2 , C(O)NH 2 , C(O)NHR 18 , C(O)NHR 18 ,
  • R is alkyl, alkenyl, alkynyl, phenyl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl.
  • Another embodiment of this invention therefore pertains to a cationic lipid or mixtures thereof, having Formula (I) Formula (I)
  • R and R are independently R ;
  • R and R taken together with the atoms to which they are attached, are heterocycloalkyl or heteroaryl;
  • L 1 is O, or OC(O);
  • L is O, or OC(O);
  • R and R is H, and the other is C 14 -C 20 -alkenyl; or
  • R and R are independently C 14 -C 20 -alkenyl or C 14 -C 20 -alkyl; or
  • R is alkyl, which is unsubstituted or substituted with R ;
  • R 6 is R 7 ;
  • R is phenyl
  • each foregoing cyclic moiety is independently unsubstituted or substituted with one or more R 1 ⁇ N(R 1 ⁇ 2 , or OH;
  • R is R , R , or R ;
  • R is phenyl
  • R is heteroaryl
  • R is alkyl, which is unsubstituted or substituted with one or two of independently selected R 16 , or N(R 16 ) 2 ;
  • R is alkyl, or R ; and R is heterocycloalkyl.
  • R 1 and R 2 are each independently R 5 .
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heteroaryl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are pyrrolidinyl, piperazinyl, 1 ,4-diazepanyl, piperidinyl, or morpholinyl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are pyrrolidinyl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are imidazolyl.
  • One embodiment of this invention pertains to compounds of Formula (I), wherein L 1 and L 2 are O. In another embodiment of Formula (I), L 1 and L 2 are OC(O). In another embodiment of Formula (I), L 1 and L 2 are (O)CO. In another embodiment of Formula (I), one of L 1 and L 2 is O, and the other is OC(O).
  • One embodiment of this invention pertains to compounds of Formula (I), wherein one of R 3 and R 4 is H, and the other is C 14 -C 20 -alkenyl.
  • one of R 3 and R 4 is H, and the other is C 14 -C 20 -alkyl.
  • R 3 and R 4 are C 1 4-C2o-alkenyl.
  • R 3 and R 4 are C14-C20- alkyl.
  • one of R 3 and R 4 is C 14 -C 20 -alkenyl, and the other is C 14 -C 20 -alkyl.
  • R 3 and R 4 combine to form CR 20 R 21 , wherein R 20 and R 21 are C 14 -C 20 -alkenyl. In another embodiment of Formula (I), R 3 and R 4 combine to form CR 20 R 21 , wherein R 20 and R 21 are C 14 -C 20 -alkyl.
  • One embodiment of this invention pertains to compounds of Formula (I), wherein R 5 is alkyl, which is unsubstituted. In another embodiment of Formula (I), R 5 is alkyl, which is substituted with R 6 . In another embodiment of Formula (I), R 6 is phenyl which is unsubstituted.
  • One embodiment of Formula (I) pertains to compounds wherein all foregoing cyclic moieties are unsubstituted. In another embodiment of Formula (I), one or more cyclic moieties are substituted. In another embodiment of Formula (I), one or more cyclic moieties are substituted with one or more R , N(R ) 2 , or OH.
  • One embodiment of this invention pertains to compounds of Formula (I) wherein R 1 and R 2 , taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 and L 2 are O; and R 3 and R 4 are C 14 -C 20 -alkenyl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heteroaryl; L 1 and L 2 are O; and R 3 and R 4 are C 14 -C 20 -alkenyl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 and L 2 are O; and R 3 and R 4 are C 14 -C 20 -alkenyl; wherein the heterocycloalkyl is substituted with R 11 ; R 11 is R 15 ; and R 15 is alkyl which is unsubstituted.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 and L 2 are O; and R 3 and R 4 are C 14 -C 20 -alkenyl; wherein the heterocycloalkyl is substituted with
  • R 11 ; R 11 is R 12 ; and R 12 is phenyl which is unfused.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 and L 2 are O; and R 3 and R 4 are C 14 -C 20 -alkenyl; wherein the heterocycloalkyl is substituted with R 11 ; R 11 is R 13 ; and R 13 is heteroaryl which is unfused.
  • R 1 and R 2 are each independently R 5 ; wherein one R 5 is alkyl which is unsubstituted, and the other R 5 is alkyl which is substituted with R 6 ; R 6 is R 7 ; R 7 is phenyl which is unfused; L 1 and L 2 are O; and R 3 and R 4 are C 14 -C 20 -alkenyl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 and L 2 are O; and R 3 and R 4 are C 14 -C 20 -alkenyl; wherein the heterocycloalkyl is substituted with R 11 ; R 11 is R 15 ; R 15 is alkyl which is substituted with N(R 16 ) 2 ; and R 16 is alkyl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 and L 2 are O; and R 3 and R 4 are C 14 -C 20 -alkenyl; wherein the heterocycloalkyl is substituted with R 11 ; R 11 is R 15 ; R 15 is alkyl which is substituted with R 16 ; R 16 is R 17 ; and R 17 is heterocycloalkyl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 and L 2 are O; and R 3 and R 4 are C 14 -C 20 -alkenyl; wherein the heterocycloalkyl is substituted with N(R ⁇ )2; R 11 is R 15 ; and R 15 is alkyl which is unsubstituted.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 and L 2 are O; and one of R 3 and R 4 is H, and the other is C14-C20- alkenyl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 and L 2 are OC(O); and R 3 and R 4 are C 14 - C 20 -alkenyl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 is O; L 2 is OC(O); and R 3 and R 4 are C 14 - C 20 -alkenyl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 and L 2 are O; and R 3 and R 4 combine to form CR 20 R 21 ; wherein R 20 and R 21 are C 14 -C 20 -alkenyl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 and L 2 are O; and R 3 and R 4 are C 14 -C 20 -alkyl.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 and L 2 are O; and R 3 and R 4 are C 14 -C 20 -alkenyl; wherein the heterocycloalkyl is substituted OH.
  • R 1 and R 2 taken together with the atoms to which they are attached, are heterocycloalkyl; L 1 and L 2 are O; and R 3 is C 1 4-C2o-alkyl and R 4 is C 14 -C 20 -alkenyl.
  • Still another embodiment pertains to compounds of Formula I which are l-(2,3- bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine, l-(2,3-bis((9Z,12Z)-octadeca- 9,12-dienyloxy)propyl)-1H-imidazole, l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)- 4-methylpiperazine, l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)-4-methyl-1,4- diazepane, l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)-4-phenylpiperazine, l-(2,3- bis((9Z,12Z)
  • Still another embodiment pertains to compounds of this invention wherein one or more cationic lipids are chosen from l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)-4- methylpiperazine, N-(2-(4-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)piperazin-1- yl)ethyl)-N,N-dimethylamine, l-((2S)-2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)-4- methylpiperazine, l-((2R)-2,3-bis((9Z,12Z)-octadeca-9
  • Still another embodiment pertains to compounds of this invention wherein one or more cationic lipids are chosen from l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)-4-
  • a further embodiment pertains to particles comprising one or more cationic lipid(s) having Formula I.
  • a further embodiment pertains to particles comprising one or more cationic lipid(s) having Formula I and one or more therapeutic agents.
  • said therapeutic agent is a nucleic acid encoded with a product of interest, including but not limited to, RNA, antisense oligonucleotide, a DNA, a plasmid, a ribosomal RNA (rRNA), a micro RNA (miRNA), transfer RNA (tRNA), a small inhibitory RNA (siRNA), small nuclear RNA (snRNA), antigens, fragments thereof, proteins, peptides, and small-molecules.
  • RNA nucleic acid encoded with a product of interest, including but not limited to, RNA, antisense oligonucleotide, a DNA, a plasmid, a ribosomal RNA (rRNA), a micro RNA (miRNA), transfer RNA (tRNA), a small inhibitory RNA (siRNA), small nuclear RNA (snRNA
  • a further embodiment pertains to nanoparticles comprising one or more cationic lipid(s) having Formula I.
  • a further embodiment pertains to nanoparticles comprising one or more cationic lipid(s) having Formula I and one or more therapeutic agents.
  • said therapeutic agent is a nucleic acid encoded with a product of interest, including but not limited to, RNA, antisense oligonucleotide, a DNA, a plasmid, a ribosomal RNA (rRNA), a micro RNA (miRNA), transfer RNA (tRNA), a small inhibitory RNA (siRNA), small nuclear RNA (snRNA), antigens, fragments thereof, proteins, peptides, and small-molecules.
  • RNA nucleic acid encoded with a product of interest, including but not limited to, RNA, antisense oligonucleotide, a DNA, a plasmid, a ribosomal RNA (rRNA), a micro RNA (miRNA), transfer RNA (tRNA), a small inhibitory RNA (siRNA), small nuclear RNA (
  • a still further embodiment pertains to Cationic-Based Lipid Encapsulation Systems (CaBLES) comprising non-cationic lipid(s), polyethylene glycol (PEG)-lipid conjugate(s) and cationic lipid(s) having Formula I.
  • CaBLES Cationic-Based Lipid Encapsulation Systems
  • a still further embodiment pertains to Cationic-Based Lipid Encapsulation Systems (CaBLES) comprising one or more cationic lipids having Formula (I)
  • R and R are independently H, cycloalkyl, cycloalkenyl or R ; or
  • R and R taken together with the atoms to which they are attached, are heterocycloalkyl or heteroaryl;
  • IT is O, OC(O) or (O)CO
  • R and R is H, and the other is C 14 -C 20 -alkenyl or C 14 -C 20 -alkyl; or
  • R and R are independently C 14 -C 20 -alkenyl or C 14 -C 20 -alkyl; or
  • R and R combine to form CR R , wherein R is H and R is C 14 -C 20 -alkenyl or
  • R and R are the same or are different and are C 14 -C 20 -alkenyl, C 14 -C 20 - alkyl, or (CH 2 O)-C 14 -C 2 O alkenyl;
  • R is alkyl, which is unsubstituted or substituted with one or more R , OR , SR , S(O)R 6 , SO 2 R 6 , C(O)R 6 , CO(O)R 6 , OC(O)R 6 , OC(O)OR 6 , NH 2 , NHR 6 , N(R 6 ) 2 , NHC(O)R 6 , NR 6 C(O)R 6 , NHS(O) 2 R 6 , N R 6 S(O) 2 R 6 , NHC(O)OR 6 , NR 6 C(O)OR 6 , NHC(O)NH 2 ,
  • R is phenyl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is heteroaryl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is alkyl, alkenyl or alkynyl
  • each foregoing cyclic moiety is independently unsubstituted or substituted with one or more R 11 , OR 11 , SR 11 , S(O)R 11 , SO 2 R 11 , C(O)R 11 , CO(O)R 11 , OC(O)R 11 , OC(O)OR 11 , NH 2 , NHR 11 , N(R 1 1 ) 2 , NHC(O)R 11 , NR 11 C(O)R 11 , NHS(O) 2 R 11 , N R 11 S(O) 2 R 11 , NHC(O)OR 11 , NR 11 C(O)OR 11 , NHC(O)NH 2 , NHC(O)NHR 11 , NHC(O)N(R 1 1 ) 2 , NR 11 C(O)NHR 11 , NR 11 C(O)N(R 1 ⁇ 2 , C(O)NH 2 , C(O)NHR 11 , C(O)N(R 11
  • R is R , R , R , or R ;
  • R is phenyl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused
  • R is heteroaryl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is alkyl, alkenyl or alkynyl, each of which is unsubstituted or substituted with one or two of independently selected R 16 , OR 16 , SR 16 , S(O) 2 R 16 , C(O)OH, NH 2 , NHR 16 N(R 16 ) 2 , C(O)R 16 , C(O)NH 2 , C(O)NHR 16 , C(O)N(R 16 ) 2 , NHC(O)R 16 , NR 16 C(O)R 16 , NHC(O)OR 16 , NR 16 C(O)OR 16 , OH, F, Cl, Br or I;
  • R is alkyl, alkenyl, alkynyl, or R ;
  • R is phenyl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R , R , R , and R are independently unsubstituted or substituted with one or more R 18 , OR 18 , SR 18 , S(O)R 18 , SO 2 R 18 , C(O)R 18 , CO(O)R 18 , OC(O)R 18 , OC(O)OR 18 , NH 2 , NHR 18 , N(R 18 ) 2 , NHC(O)R 18 , NR 18 C(O)R 18 , NHS(O) 2 R 18 , NR 18 S(O) 2 R 18 , NHC(O)OR 18 , NR 18 C(O)OR 18 , NHC(O)NH 2 , NHC(O)NHR 18 , NHC(O)N(R 18 ) 2 , NR 18 C(O)NHR 18 , NR 18 C(O)N(R 18 ) 2 , C(O)NH 2 , C(O)NHR 18 ,
  • R is alkyl, alkenyl, alkynyl, phenyl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl;
  • non-cationic lipids one or more non-cationic lipids, and one or more polyethylene glycol-lipid conjugates.
  • Lipid-Based Particles of the present invention are defined as CaBLES which further comprise one or more therapeutic agent(s).
  • Therapeutic agents that can be delivered with CaBLES include RNA, antisense oligonucleotide, a DNA, a plasmid, a ribosomal RNA (rRNA), a micro RNA (miRNA), transfer RNA (tRNA), a small inhibitory RNA (siRNA), small nuclear RNA (snRNA), chimeric nucleic acids, an antigen, fragments thereof, a protein, a peptide, small-molecules, or mixtures thereof.
  • This invention describes delivery of RNA's such as small inhibitory RNA or microRNA.
  • the nucleic acid can have varying lengths (10-200 bps) and structures (hairpins, single/double strands, bulges, nicks/gaps, mismatches) and processed in the cell to provide active gene silencing.
  • a double-stranded siRNA can have the same number of nucleotides on each strand (blunt ends) or asymmetric ends (overhangs). The overhang of 1-2 nucleotides can be present on the sense and/or the antisense strand, as well as present on the 5'- and/ or the 3 '-ends of a given strand.
  • the CaBLES and/or the Lipid-Based Particle formulation can have a ligand attached, such as a targeting ligand or a chelating moiety for complexing calcium.
  • a ligand attached such as a targeting ligand or a chelating moiety for complexing calcium.
  • the cationic lipids of Formula I maintains a positive charge.
  • the ligand that is attached has a positive charge.
  • Suitable ligands include, but are not limited to, a compound or device with a reactive functional group and include lipids, amphipathic lipids, carrier compounds, bioaffinity compounds, biomaterials, biopolymers, biomedical devices, analytically detectable compounds, therapeutically active compounds, enzymes, peptides, proteins, antibodies, immune stimulators, radiolabels, fluorogens, biotin, drugs, haptens, DNA, RNA, polysaccharides, liposomes, virosomes, micelles, immunoglobulins, functional groups, other targeting moieties, or toxins.
  • a targeting ligand is conjugated to the periphery of the PEG-lipid in a Lipid-Based Particle formulation.
  • the targeting moiety is a ligand of a receptor present on a target cell and the receptor is preferentially expressed by the target cell versus a non-target cell.
  • the targeting moiety is an antibody or fragments thereof.
  • the targeting moiety is a small protein, or peptide. In another aspect, the targeting moiety is a small-molecule.
  • these Lipid-Based Particles are nanoparticles and have mean diameter sizes of about 50-300 nm, of which 50-250 nm is preferred and 50-200 nm is most preferred.
  • a further embodiment pertains to CaBLES or Lipid-Base Particles wherein the PEG lipid conjugate is about 0.1-20 weight/weight % of total lipid in particle, the non-cationic lipid is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the cationic lipid is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to CaBLES or Lipid-Base Particles wherein the PEG lipid conjugate is about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the cationic lipid is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a pharmaceutical composition
  • a pharmaceutical composition comprising a Lipid- Based Particle and a pharmaceutically acceptable carrier.
  • a further embodiment pertains to a pharmaceutical composition, wherein the Lipid- Based Particle comprises cholesterol, DSPC, l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, one or more PEG-lipid conjugates, and one or more nucleic acids.
  • the Lipid- Based Particle comprises cholesterol, DSPC, l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, one or more PEG-lipid conjugates, and one or more nucleic acids.
  • a further embodiment pertains to a pharmaceutical composition, wherein the (PEG)- lipid conjugates are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight %
  • l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugate is N-(2,3- dimyristyloxypropyl)carbamate polyethyleneglycol-2000 methyl ether, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • the PEG-lipid conjugate is N-(2,3- dimyristyloxypropyl)carbamate polyethyleneglycol-2000 methyl ether
  • the therapeutic agent is siRNA.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-(2,3- dimyristyloxypropyl)carbamate polyethyleneglycol-2000 methyl ether is about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugate is N-(carbonyl- methoxypolyethyleneglycol-2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • the PEG-lipid conjugate is N-(carbonyl- methoxypolyethyleneglycol-2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-distearoyl-sn-glycero-3-phosphoethanolamine is about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugate is N-(carbonyl- methoxypolyethyleneglycol-2000)-1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • the PEG-lipid conjugate is N-(carbonyl- methoxypolyethyleneglycol-2000)-1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-(carbonyl- methoxypolyethyleneglycol-2000)-1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine is about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5- 60 weight/weight % of total lipid in particle.
  • the N-(carbonyl- methoxypolyethyleneglycol-2000)-1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine is about 0.1-20 weight/weight % of total lipid in particle
  • the DSPC is about 1-30 weight/weight % of total lipid in particle
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugate is N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-dimyristoyl-sn-glycero-3-phosphoethanolamine, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • the PEG-lipid conjugate is N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-dimyristoyl-sn-glycero-3
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-(carbonyl- methoxypolyethyleneglycol-2000)-l ,2-dimyristoyl-sn-glycero-3-phosphoethanolamine is about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5- 60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugate is 1 ,2-distearoyl-sn-glycerol- methoxypolyethyleneglycol-2000, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • the PEG-lipid conjugate is 1 ,2-distearoyl-sn-glycerol- methoxypolyethyleneglycol-2000
  • the therapeutic agent is siRNA.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the 1 ,2-distearoyl- sn-glycerol-methoxypolyethyleneglycol-2000 is about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca- 9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3bis((9Z,12Z)-octadeca-9,12-
  • the PEG-lipid conjugate is 1 ,2-dipalmitoyl-sn-glycerol- methoxypolyethyleneglycol-2000, and the therapeutic agent is siRNA.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the 1 ,2-dipalmitoyl- sn-glycerol-methoxypolyethyleneglycol-2000 is about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca- 9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugate is 1 ,2-dimyristoyl-sn-glycerol- methoxypolyethyleneglycol-2000, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • the PEG-lipid conjugate is 1 ,2-dimyristoyl-sn-glycerol- methoxypolyethyleneglycol-2000
  • the therapeutic agent is siRNA.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the 1 ,2-dimyristoyl- sn-glycerol-methoxypolyethyleneglycol-2000 is about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca- 9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugate is N-[3,4-bis(tetradecyloxy)butyl]- 2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,74,77,80,83,86,89,92, 95,98,101,104,107,110,113,116,119,122,125,128,131,134,137- hexatetracontaoxanonatriacontahectan-139-amide, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the DSPC is about 1-30 weight/weight % of total lipid in particle
  • the cholesterol is about 5-45 weight/weight % of total lipid in particle
  • 29 bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugate is N-[3,4-bis(hexadecyloxy)butyl]- 2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,74,77,80,83,86,89,92, 95,98,101,104,107,110,113,116,119,122,125,128,131,134,137- hexatetracontaoxanonatriacontahectan-139-amide, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-[3,4- bis(hexadecyloxy)butyl] -
  • 2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,74,77,80,83,86,89,92, 95,98,101,104,107,110,113,116,119,122,125,128,131,134,137- hexatetracontaoxanonatriacontahectan- 139-amide is about 0.1-20 weight/weight % of total lipid in particle
  • the DSPC is about 1-30 weight/weight % of total lipid in particle
  • the cholesterol is about 5-45 weight/weight % of total lipid in particle
  • the l-(2,3- bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a pharmaceutical composition, wherein the Lipid- Based Particle comprises, cholesterol, SPC, l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, one or more PEG-lipid conjugates, and one or more nucleic acids.
  • the Lipid- Based Particle comprises, cholesterol, SPC, l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, one or more PEG-lipid conjugates, and one or more nucleic acids.
  • a further embodiment pertains to a pharmaceutical composition, wherein the (PEG)- lipid conjugates are about 0.1-20 weight/weight % of total lipid in particle, the SPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • the (PEG)- lipid conjugates are about 0.1-20 weight/weight % of total lipid in particle
  • the SPC is about 1-30 weight/weight % of total lipid in particle
  • the cholesterol is about 5-45 weight/weight % of total lipid in particle
  • l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and SPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12-
  • the PEG-lipid conjugate is 1 ,2-dipalmitoyl-sn-glycerol- methoxypolyethyleneglycol-2000 and , and the therapeutic agent is siRNA.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the 1 ,2-dipalmitoyl- sn-glycerol-methoxypolyethyleneglycol-2000 is about 0.1-20 weight/weight % of total lipid in particle, the SPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca- 9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • the 1 ,2-dipalmitoyl- sn-glycerol-methoxypolyethyleneglycol-2000 is about 0.1-20 weight/weight % of total lipid in particle
  • the SPC is about 1-30 weight/weight % of total lipid in particle
  • the cholesterol is about 5-45 weight/weight % of total lipid in particle
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugates are 1 ,2-dimyristoyl-sn-glycerol- methoxypolyethyleneglycol-2000 and 1 ,2-dipalmitoyl-sn-glycerol- methoxypolyethyleneglycol-2000, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • the PEG-lipid conjugates are 1 ,2-dimyristoyl-sn-glycerol- meth
  • a further embodiment pertains to a Lipid-Based Particle, wherein the 1 ,2-dimyristoyl- sn-glycerol-methoxypolyethyleneglycol-2000 and 1 ,2-dipalmitoyl-sn-glycerol- methoxypolyethyleneglycol-2000 are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5- 45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugates are 1 ,2-dimyristoyl-sn-glycerol- methoxypolyethyleneglycol-2000 and 1 ,2-distearoyl-sn-glycerol- methoxypolyethyleneglycol-2000, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • the PEG-lipid conjugates are 1 ,2-dimyristoyl-sn-glycerol- meth
  • a further embodiment pertains to a Lipid-Based Particle, wherein the 1 ,2-dimyristoyl- sn-glycerol-methoxypolyethyleneglycol-2000 and 1 ,2-distearoyl-sn-glycerol- methoxypolyethyleneglycol-2000 are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5- 45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugates are 1 ,2-dipalmitoyl-sn-glycerol- methoxypolyethyleneglycol-2000 and 1 ,2-distearoyl-sn-glycerol- methoxypolyethyleneglycol-2000, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • the PEG-lipid conjugates are 1 ,2-dipalmitoyl-sn-glycerol- meth
  • a further embodiment pertains to a Lipid-Based Particle, wherein the 1 ,2-dipalmitoyl- sn-glycerol-methoxypolyethyleneglycol-2000 and 1 ,2-distearoyl-sn-glycerol- methoxypolyethyleneglycol-2000 are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5- 45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugates are N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-dimyristoyl-sn-glycero-3-phosphoethanolamine and N-(carbonyl-methoxypolyethyleneglycol-2000)-1,2-dipalmitoyl-sn-glycero-3- phosphoethanolamine, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-dimyristoyl-sn-glycero-3-phosphoethanolamine and N-(carbonyl-methoxypolyethyleneglycol-2000)-1,2-dipalmitoyl-sn-glycero-3- phosphoethanolamine are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugates are N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-distearoyl-sn-glycero-3-phosphoethanolamine and N- (carbonyl-methoxypolyethyleneglycol-2000)-1,2-dipalmitoyl-sn-glycero-3- phosphoethanolamine, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-distearoyl-sn-glycero-3-phosphoethanolamine and N- (carbonyl-methoxypolyethyleneglycol-2000)-1,2-dipalmitoyl-sn-glycero-3- phosphoethanolamine are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugates are N-[3,4-bis(hexadecyloxy)butyl]- 2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,74,77,80,83,86,89,92, 95,98,101,104,107,110,113,116,119,122,125,128,131,134,137- hexatetracontaoxanonatriacontahectan- 139-amide and 1 ,2-distearoyl-sn-glycerol- methoxypolyethyleneglycol
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-[3,4- bis(hexadecyloxy)butyl] - 2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,74,77,80,83,86,89,92, 95,98,101,104,107,110,113,116,119,122,125,128,131,134,137- hexatetracontaoxanonatriacontahectan- 139-amide and 1 ,2-distearoyl-sn-glycerol- methoxypolyethyleneglycol-2000 are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5- 45 weight/weight % of total lipid in particle, and the l-(2,3-bis((
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugates are N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-dimyristoyl-sn-glycero-3-phosphoethanolamine and N-(carbonyl-methoxypolyethyleneglycol-2000)-1,2-distearoyl-sn-glycero-3- phosphoethanolamine, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-dimyristoyl-sn-glycero-3-phosphoethanolamine and N-(carbonyl-methoxypolyethyleneglycol-2000)-1,2-distearoyl-sn-glycero-3- phosphoethanolamine are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugates are 1 ,2-dimyristoyl-sn-glycerol- methoxypolyethyleneglycol-2000 and N-(carbonyl-methoxypolyethyleneglycol-2000)-l ,2- distearoyl-sn-glycero-3-phosphoethanolamine, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • the PEG-lipid conjugates are 1 ,2-dimyristoy
  • a further embodiment pertains to a Lipid-Based Particle, wherein the 1 ,2-dimyristoyl- sn-glycerol-methoxypolyethyleneglycol-2000 and N-(carbonyl-methoxypolyethyleneglycol- 2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3- bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugates are 1 ,2-distearoyl-sn-glycerol- methoxypolyethyleneglycol-2000 and N-(carbonyl-methoxypolyethyleneglycol-2000)-l ,2- dimyristoyl-sn-glycero-3-phosphoethanolamine, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • the PEG-lipid conjugates are 1 ,2-distearoy
  • a further embodiment pertains to a Lipid-Based Particle, wherein the 1 ,2-distearoyl- sn-glycerol-methoxypolyethyleneglycol-2000 and N-(carbonyl-methoxypolyethyleneglycol- 2000)-1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3- bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugates are N-[3,4-bis(hexadecyloxy)butyl]-
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-[3,4- bis(hexadecyloxy)butyl]-
  • the cholesterol is about 5-45 weight/weight % of total lipid in particle
  • the l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5-
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugates are N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-dimyristoyl-sn-glycero-3-phosphoethanolamine and
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-dimyristoyl-sn-glycero-3-phosphoethanolamine and 1,2-dipalmitoyl-sn-glycerol-methoxypolyethyleneglycol-2000 are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3- bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a pharmaceutical composition, wherein the Lipid- Based Particle comprises, cholesterol, DSPC, l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)-4-methylpiperazine, one or more PEG-lipid conjugates, and one or more nucleic acids.
  • the Lipid- Based Particle comprises, cholesterol, DSPC, l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)-4-methylpiperazine, one or more PEG-lipid conjugates, and one or more nucleic acids.
  • a further embodiment pertains to a pharmaceutical composition, wherein the (PEG)- lipid conjugates are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)-4- methylpiperazine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)-4-methylpiperazine, the PEG-lipid conjugate is N-(2,3- dimyristyloxypropyl)carbamate polyethyleneglycol-2000 methyl ether, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)-4-methylpiperazine
  • the PEG-lipid conjugate is N-(2,3- dimyristyloxypropyl)carbamate polyethyleneglycol-2000 methyl ether
  • the therapeutic agent is siRNA.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-(2,3- dimyristyloxypropyl)carbamate polyethyleneglycol-2000 methyl ether is about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)-4-methylpiperazine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a pharmaceutical composition, wherein the Lipid- Based Particle comprises, cholesterol, DSPC, l-((2S)-2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, one or more PEG-lipid conjugates, and one or more nucleic acids.
  • the Lipid- Based Particle comprises, cholesterol, DSPC, l-((2S)-2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, one or more PEG-lipid conjugates, and one or more nucleic acids.
  • a further embodiment pertains to a pharmaceutical composition, wherein the (PEG)- lipid conjugates are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and l-((2S)-2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-((2S)-2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugate is N-(2,3- dimyristyloxypropyl)carbamate polyethyleneglycol-2000 methyl ether, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-((2S)-2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • the PEG-lipid conjugate is N-(2,3- dimyristyloxypropyl)carbamate polyethyleneglycol-2000 methyl ether
  • the therapeutic agent is siRNA.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-(2,3- dimyristyloxypropyl)carbamate polyethyleneglycol-2000 methyl ether is about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-((2S)-2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a pharmaceutical composition, wherein the Lipid-
  • Based Particle comprises, cholesterol, DSPC, l-((2R)-2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, one or more PEG-lipid conjugates, and one or more nucleic acids.
  • a further embodiment pertains to a pharmaceutical composition, wherein the (PEG)- lipid conjugates are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and l-((2R)-2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l-((2R)-2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine, the PEG-lipid conjugate is N-(2,3- dimyristyloxypropyl)carbamate polyethyleneglycol-2000 methyl ether, and the therapeutic agent is siRNA.
  • the non-cationic lipids are cholesterol and DSPC
  • the cationic lipid is l-((2R)-2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)pyrrolidine
  • the PEG-lipid conjugate is N-(2,3- dimyristyloxypropyl)carbamate polyethyleneglycol-2000 methyl ether
  • the therapeutic agent is siRNA.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-(2,3- dimyristyloxypropyl)carbamate polyethyleneglycol-2000 methyl ether is about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l-((2R)-2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a pharmaceutical composition, wherein the Lipid-
  • Based Particle comprises, cholesterol, DSPC, l- ⁇ 2-[(9E, 12E)-octadeca-9,12-dienyloxy]-3- [(9Z,12Z)-octadeca-9,12-dienyloxy]propyl ⁇ pyrrolidine, one or more PEG-lipid conjugates, and one or more nucleic acids.
  • a further embodiment pertains to a pharmaceutical composition, wherein the (PEG)- lipid conjugates are about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and l- ⁇ 2-[(9E, 12E)-octadeca-9,12-dienyloxy]-3-[(9Z,12Z)- octadeca-9,12-dienyloxy]propyl ⁇ pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l- ⁇ 2-[(9E, 12E)-octadeca-9,12- dienyloxy]-3-[(9Z,12Z)-octadeca-9,12-dienyloxy]propyl ⁇ pyrrolidine, the PEG-lipid
  • conjugate is l ⁇ -dipalmitoyl-sn-glycerol-methoxypolyethyleneglycol ⁇ OOO, and the therapeutic agent is siRNA.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the 1 ,2-dipalmitoyl- sn-glycerol-methoxypolyethyleneglycol-2000 is about 0.1-20 weight/weight % of total lipid in particle, the DSPC is about 1-30 weight/weight % of total lipid in particle, the cholesterol is about 5-45 weight/weight % of total lipid in particle, and the l- ⁇ 2-[(9E, 12E)-octadeca- 9,12-dienyloxy]-3-[(9Z,12Z)-octadeca-9,12-dienyloxy]propyl ⁇ pyrrolidine is about 5-60 weight/weight % of total lipid in particle.
  • a further embodiment pertains to Lipid-Based Particles, wherein the ratio of one or more (PEG)-lipid conjugates, one or more non-cationic lipids, and one or more cationic lipids of Formula (I), to one or more therapeutic agents is between about 50:1 to about 5:1.
  • the A further embodiment pertains to Lipid-Based Particles, wherein the ratio of one or more (PEG)-lipid conjugates, one or more non-cationic lipids, and one or more cationic lipids of Formula (I), to one or more therapeutic agents is between about 30:1 to about 10:1.
  • functional CaBLES comprising one or more (PEG)- lipid conjugates, one or more non-cationic lipids, and one or more cationic lipids of Formula 1 effectively encapsulate nucleic acids, such as siRNA, with efficiencies from about 50- 100%.
  • functional CaBLES comprising one or more (PEG)- lipid conjugates, one or more non-cationic lipids, and one or more cationic lipids of Formula 1 effectively encapsulate nucleic acids, such as siRNA, with efficiencies from about 80- 100%.
  • functional CaBLES comprising one or more (PEG)- lipid conjugates, one or more non-cationic lipids, and one or more cationic lipids chosen from l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine, l-(2,3-bis((9Z,12Z)- octadeca-9,12-dienyloxy)propyl)-1H-imidazole, l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)-4-methylpiperazine, l-(2,3-bis((9Z,12Z)-octadeca-9,12- dienyloxy)propyl)-4-methyl-1,4-diazepane, l-(2,3-bis((9Z,12Z)-octadeca-9,12- dieny
  • functional CaBLES comprising one or more (PEG)- lipid conjugates, one or more non-cationic lipids, and one or more cationic lipids chosen from l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine, l-(2,3-bis((9Z,12Z)- octadeca-9,12-dienyloxy)propyl)-1H-imidazole, l-(2,3-bis((9Z,12Z)-octadeca-9,12-
  • a further embodiment pertains to examples of non-cationic lipids that are useful for the practice of this invention which include, but are not limited to, cholesterol, cholesterol sulfate, ceramide, sphingomyelin, lecithin, sphingomyelin, egg sphingomyelin, milk sphingomyelin; egg phosphatidylcholine, hydrogenated egg phosphatidylcholine, hydrogenated soybean phosphatidylethanolamine, egg phosphatidylethanolamine, hydrogenated soybean phosphatidylcholine, soybean phosphatidylcholine, 1 ,2-dilauroyl-sn- glycerol, 1 ,2-dimyristoyl-sn-glycerol, 1 ,2-dipalmitoyl-sn-glycerol, 1 ,2-distearoyl-sn-glycerol, 1 ,2-dilauroyl-sn-glycero-3-phosphatidic acid
  • PEG-lipid conjugates that are useful for the practice of this invention which include, but are not limited to, 2-(tetradecyloxy)-1-
  • PEG-lipid conjugates are described in, e.g., US App. No. 61/095,748, which was filed
  • PEG-lipid conjugates are described in, e.g., US App. No. 61/095,769, which was filed on September 10, 2008 and is incorporated herein by reference.
  • a still further embodiment pertains to combinations of polyethylene glycol (PEG)- lipid conjugates which are useful for the practice of this invention, wherein two PEG-lipid conjugates are chosen from N-(carbonyl-methoxypolyethyleneglycol-2000)-1,2-dimyristoyl- sn-glycero-3-phosphoethanolamine , N-(carbonyl-methoxypolyethyleneglycol-2000)- 1 ,2- dipalmitoyl-sn-glycero-3 -phosphoethanolamine, N-(carbonyl-methoxypolyethyleneglycol- 200O)-1 ,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1 ,2-distearoyl-sn-glycerol- methoxypolyethyleneglycol-2000, 1,2-dimyristoyl-sn-glycerol-methoxypolyethyleneglycol- 2000, 1,2-dipalmitoyl-sn-glycerol-
  • a still further embodiment pertains to combinations of polyethylene glycol (PEG)- lipid conjugates which are useful for the practice of this invention, wherein at least one of the PEG-lipid conjugates is chosen from N-(carbonyl-methoxypolyethyleneglycol-2000)- 1,2- dimyristoyl-sn-glycero-3-phosphoethanolamine, N-(carbonyl-methoxypolyethyleneglycol- 2000)-1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine, N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-distearoyl-sn-glycero-3 -phosphoethanolamine, 1 ,2- distearoyl-sn-glycerol-methoxypolyethyleneglycol-2000, 1 ,2-dimyristoyl-sn-glycerol- methoxypolyethyleneglycol-2000, 1,2-dipalmitoyl-sn-glycerol-
  • a still further embodiment pertains to combinations of polyethylene glycol (PEG)- lipid conjugates which are useful for the practice of this invention and include 1,2- dimyristoyl-sn-glycerol-methoxypolyethyleneglycol-2000 and N-(carbonyl-
  • Particles comprises about 2 to about 60 weight/weight percent of total lipid in the particle.
  • the non-cationic lipids of the Cables and Lipid-Based Particles comprises about 5 to about 90 weight/weight percent of total lipid in the particle.
  • the PEG-lipid conjugates of the CaBLES and Lipid- Based Particles comprises from 0.1 to about 20 weight/weight percent of total lipid in the particle.
  • Still another embodiment pertains to a method of treating cancer in a mammal comprising administering thereto a Lipid-Based Particle.
  • Still another embodiment comprises methods of treating cancer in a mammal comprising administering thereto a Lipid-Based Particle comprising one or more cationic lipids having Formula (I)
  • R and R are independently H, cycloalkyl, cycloalkenyl or R ; or
  • R and R taken together with the atoms to which they are attached, are heterocycloalkyl or heteroaryl;
  • L 1 is O, OC(O) or (O)CO
  • L 2 is O, OC(O) or (O)CO
  • R and R is H, and the other is C 14 -C 20 -alkenyl or C 14 -C 20 -alkyl; or
  • R and R are independently C 14 -C 20 -alkenyl or C 14 -C 20 -alkyl; or
  • R and R combine to form CR R , wherein R is H and R is C 14 -C 20 -alkenyl or
  • C 14 -C 20 -alkyl or R and R are the same or are different and are C 14 -C 20 -alkenyl, C 14 -C 20 - alkyl, or (CH 2 O)-C 14 -C 2 O alkenyl;
  • R is alkyl, which is unsubstituted or substituted with one or more R , OR , SR , S(O)R 6 , SO 2 R 6 , C(O)R 6 , CO(O)R 6 , OC(O)R 6 , OC(O)OR 6 , NH 2 , NHR 6 , N(R 6 ) 2 , NHC(O)R 6 , NR 6 C(O)R 6 , NHS(O) 2 R 6 , N R 6 S(O) 2 R 6 , NHC(O)OR 6 , NR 6 C(O)OR 6 , NHC(O)NH 2 , NHC(O)NHR 6 , NHC(O)N(R 6 ) 2 , NR 6 C(O)NHR 6 , NR 6 C(O)N(R 6 ) 2 , C(O)NH 2 , C(O)NHR 6 , C(O)N(R 6 ) 2 , C(O)
  • r R> 6 i • s r R> 7 , r R> 8 , r R> 9 , or r R> 10 ;
  • R is phenyl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is heteroaryl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is alkyl, alkenyl or alkynyl
  • each foregoing cyclic moiety is independently unsubstituted or substituted with one or more R 11 , OR 11 , SR 11 , S(O)R 11 , SO 2 R 11 , C(O)R 11 , CO(O)R 11 , OC(O)R 11 , OC(O)OR 11 , NH 2 , NHR 11 , N(R 1 1 ) 2 , NHC(O)R 11 , NR 11 C(O)R 11 , NHS(O) 2 R 11 , N R 11 S(O) 2 R 11 , NHC(O)OR 11 , NR 11 C(O)OR 11 , NHC(O)NH 2 , NHC(O)NHR 11 , NHC(O)N(R 1 1 ) 2 , NR 11 C(O)NHR 11 , NR 11 C(O)N(R 1 ⁇ 2 , C(O)NH 2 , C(O)NHR 11 ,
  • R is R , R , R , or R ;
  • R is phenyl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is heteroaryl which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R is alkyl, alkenyl or alkynyl, each of which is unsubstituted or substituted with one or two of independently selected R 16 , OR 16 , SR 16 , S(O) 2 R 16 , C(O)OH, NH 2 , NHR 16 N(R 16 ) 2 , C(O)R 16 , C(O)NH 2 , C(O)NHR 16 , C(O)N(R 16 ) 2 , NHC(O)R 16 , NR 16 C(O)R 16 , NHC(O)OR 16 , NR 16 C(O)OR 16 , OH, F, Cl, Br or I;
  • R is alkyl, alkenyl, alkynyl, or R ;
  • R is phenyl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene, each of which is unfused or fused with benzene, heteroarene, cycloalkane, cycloalkene, heterocycloalkane, or heterocycloalkene;
  • R , R , R , and R are independently unsubstituted or substituted with one or more R 18 , OR 18 , SR 18 , S(O)R 18 , SO 2 R 18 , C(O)R 18 , CO(O)R 18 , OC(O)R 18 , OC(O)OR 18 , NH 2 , NHR 18 , N(R 18 ) 2 , NHC(O)R 18 , NR 18 C(O)R 18 , NHS(O) 2 R 18 , NR 18 S(O) 2 R 18 , NHC(O)OR 18 , NR 18 C(O)OR 18 , NHC(O)NH 2 , NHC(O)NHR 18 , NHC(O)N(R 18 ) 2 , NR 18 C(O)NHR 18 , NR 18 C(O)N(R 18 ) 2 , C(O)NH 2 , C(O)NHR 18 , C(O)NHR 18 ,
  • R is alkyl, alkenyl, alkynyl, phenyl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl;
  • non-cationic lipids one or more non-cationic lipids, one or more polyethylene glycol-lipid conjugates and one or more therapeutic agents.
  • Particles comprising: (a) mixing the cationic lipid(s), the non-cationic lipid(s) and the PEG- lipid conjugate(s); (b) adding the mixture of step (a) to one or more therapeutic agents; and
  • step (c) separating and purifying resulting suspension of step (b).
  • a further embodiment pertains to a method of making Lipid-Based Particles wherein the therapeutic agent is warmed to about 60° C prior to the addition of the mixture of step (a) via needle injection.
  • Lipid-Based Particles of this invention depend on recipient of treatment, disease treated and severity thereof, composition comprising it, time of administration, route of administration, duration of treatment, potency, rate of clearance and whether or not another drug is co-administered.
  • the amount of Lipid-Based Particles of this invention used to make compositions to be administered daily to a patient in a single dose or in divided doses is from about 0.001 to about 200 mg/kg body weight.
  • Single dose compositions contain these amounts or a combination of submultiples thereof.
  • One embodiment pertains to a pharmaceutical composition
  • a pharmaceutical composition comprising one or more (PEG)-lipid conjugates, one or more non-cationic lipids, one or more cationic lipids of Formula 1 , one or more therapeutic agents, and a pharmaceutically acceptable excipient.
  • Lipid-Based Particles of this invention may be administered, for example, bucally, ophthalmically, orally, osmotically, parenterally (intramuscularly, intraperintoneally intrasternally, intravenously, subcutaneously), rectally, topically, transdermally, vaginally and intraarterially as well as by intraarticular injection, infusion, and placement in the body, such as, for example, the vasculature.
  • Lipid-Based Particles may be administered with or without an excipient.
  • Excipients include, but are not limited to, encapsulators and additives such as absorption accelerators, antioxidants, binders, buffers, coating agents, coloring agents, diluents, disintegrating agents, emulsif ⁇ ers, extenders, fillers, flavoring agents, humectants, lubricants, perfumes, preservatives, propellants, releasing agents, sterilizing agents, sweeteners, solubilizers, wetting agents, mixtures thereof and the like.
  • encapsulators and additives such as absorption accelerators, antioxidants, binders, buffers, coating agents, coloring agents, diluents, disintegrating agents, emulsif ⁇ ers, extenders, fillers, flavoring agents, humectants, lubricants, perfumes, preservatives, propellants, releasing agents, sterilizing agents, sweeteners, solubilizers, wetting agents, mixtures thereof
  • Excipients for preparation of compositions comprising Lipid-Based Particles to be administered orally include, but are not limited to, agar, alginic acid, aluminum hydroxide, benzyl alcohol, benzyl benzoate, 1,3-butylene glycol, carbomers, castor oil, cellulose, cellulose acetate, cocoa butter, corn starch, corn oil, cottonseed oil, cross-povidone, diglycerides, ethanol, ethyl cellulose, ethyl laureate, ethyl oleate, fatty acid esters, gelatin, germ oil, glucose, glycerol, groundnut oil, hydroxypropylmethyl celluose, isopropanol, isotonic saline, lactose, magnesium hydroxide, magnesium stearate, malt, mannitol, monoglycerides, olive oil, peanut oil, potassium phosphate salts, potato starch, povidone, propylene glycol,
  • compositions comprising a compound having formula (I) to be administered osmotically include, but are not limited to, chlorofluorohydrocarbons, ethanol, water, mixtures thereof and the like.
  • Excipients for preparation of compositions comprising a compound having formula (I) to be administered parenterally include, but are not limited to, 1,3-butanediol, castor oil, corn oil, cottonseed oil, dextrose, germ oil, groundnut oil, liposomes, oleic acid, olive oil, peanut oil, Ringer's solution, safflower oil, sesame oil, soybean oil, U.S.P. or isotonic sodium chloride solution, water, mixtures thereof and the like.
  • Excipients for preparation of compositions comprising a compound having formula (I) to be administered rectally or vaginally include, but are not limited to, cocoa butter, polyethylene glycol, wax, mixtures thereof and the like.
  • compositions and the method of the present invention may further comprise other therapeutically active compounds as noted herein which are usually applied in the treatment of the above-mentioned pathological conditions.
  • the present invention further provides methods of using a compound, formulation, or composition of the invention in combination with one or more additional active agents.
  • Lipid-Based Particles are expected to be useful when used with: alkylating agents, angiogenesis inhibitors, antibodies, antimetabolites, antimitotics, antiproliferatives, aurora kinase inhibitors, apoptosis promoters (for example, Bcl-xL, Bcl-w and BfI-I) inhibitors, Bcr-Abl kinase inhibitors, BiTE (Bi-Specif ⁇ c T cell Engager) antibodies, biologic response modifiers, cyclin-dependent kinase inhibitors, cell cycle inhibitors, cyclooxygenase-2 inhibitors, DVD's, leukemia viral oncogene homolog (ErbB2) receptor inhibitors, growth factor inhibitors, heat shock protein (HSP)-90 inhibitors, histone deacetylase (HDAC) inhibitors, hormonal therapies, immunologicals, inhibitors of apoptosis proteins (IAP's) intercalating antibiotics, kinase inhibitors, mammalian target of
  • tyrosine kinase inhibitors 53 tyrosine kinase inhibitors, retinoids/deltoids plant alkaloids, small inhibitory ribonucleic acids (siRNA's), topoisomerase inhibitors, combinations thereof and the like.
  • a BiTE antibody is a bi-specific antibody that directs T-cells to attach cancer cells by simultaneously binding the two cells. The T-cell then attacks the target cancer cell.
  • Exemplary BiTE antibodies include adecatumumab (Micromet MT201), blinatumomab (Micromet MT103) and the like.
  • SiRNA's are molecules having endogenous RNA bases or chemically modified nucleotides. The modifications shall not abolish cellular activity, but rather impart increased stability and/or increased cellular potency. Examples of chemical modifications include phosphorothioate groups, 2'-deoxynucleotide, 2'-OCH 3 -containing ribonucleotides, 2'-F- ribonucleotides, 2'-methoxyethyl ribonucleotides or a combination thereof.
  • the siRNA can have varying lengths (10-200 bps) and structures (hairpins, single/double strands, bulges, nicks/gaps, mismatches) and processed in the cell to provide active gene silencing.
  • a double-stranded siRNA can have the same number of nucleotides on each strand (blunt ends) or asymmetric ends (overhangs). The overhang of 1-2 nucleotides can be present on the sense and/or the antisense strand, as well as present on the 5'- and/ or the 3 '-ends of a given strand.
  • Multivalent binding proteins are binding proteins comprising two or more antigen binding sites.
  • the multivalent binding protein is preferably engineered to have the three or more antigen binding sites and is generally not a naturally occurring antibody.
  • multispecific binding protein means a binding protein capable of binding two or more related or unrelated targets.
  • Dual variable domain (DVD) binding proteins are tetravalent or multivalent binding proteins binding proteins comprising two or more antigen binding sites. Such DVDs may be monospecific, i.e., capable of binding one antigen or multispecific, i.e., capable of binding two or more antigens. DVD binding proteins comprising two heavy chain DVD polypeptides and two light chain DVD polypeptides are referred to as DVD Ig. Each half of a DVD Ig comprises a heavy chain DVD polypeptide, a light chain DVD polypeptide, and two antigen binding sites. Each binding site comprises a heavy chain variable domain and a light chain variable domain with a total of 6 CDRs involved in antigen binding per antigen binding site.
  • Alkylating agents include altretamine, AMD-473, AP-5280, apaziquone, bendamustine, brostallicin, busulfan, carboquone, carmustine (BCNU), chlorambucil,
  • CLORETAZINE laromustine, VNP 4010 IM
  • cyclophosphamide decarbazine
  • estramustine fotemustine
  • glufosfamide ifosfamide
  • KW-2170 lomustine (CCNU)
  • mafosfamide melphalan
  • mitobronitol mitolactol
  • nimustine nitrogen mustard N-oxide
  • ranimustine ranimustine
  • temozolomide thiotepa
  • TREANDA biendamustine
  • treosulfan rofosfamide and the like.
  • Angiogenesis inhibitors include endothelial-specific receptor tyrosine kinase (Tie-2) inhibitors, epidermal growth factor receptor (EGFR) inhibitors, insulin growth factor-2 receptor (IGFR-2) inhibitors, matrix metalloproteinase-2 (MMP-2) inhibitors, matrix metalloproteinase-9 (MMP-9) inhibitors, platelet-derived growth factor receptor (PDGFR) inhibitors, thrombospondin analogs, vascular endothelial growth factor receptor tyrosine kinase (VEGFR) inhibitors and the like.
  • Tie-2 endothelial-specific receptor tyrosine kinase
  • EGFR epidermal growth factor receptor
  • IGFR-2 insulin growth factor-2 receptor
  • MMP-2 matrix metalloproteinase-2
  • MMP-9 matrix metalloproteinase-9
  • PDGFR platelet-derived growth factor receptor
  • VEGFR vascular endothelial growth factor receptor tyrosine
  • Antimetabolites include ALIMTA (metrexed disodium, LY231514, MTA),
  • ALKERAN (melphalan), mercaptopurine, 6-mercaptopurine riboside, methotrexate, mycophenolic acid, nelarabine, nolatrexed, ocfosfate, pelitrexol, pentostatin, raltitrexed, Ribavirin, triapine, trimetrexate, S-I, tiazofurin, tegafur, TS-I, vidarabine, UFT and the like.
  • Bcl-2 proteins inhibitors include AT-IOl ((-)gossypol), GENASENSE ® (G3139 or oblimersen (Bcl-2-targeting antisense oligonucleotide)), IPI-194, IPI-565, N-(4-(4-((4'- chloro( 1 , 1 '-biphenyl)-2-yl)methyl)piperazin- 1 -yl)benzoyl)-4-((( 1 R)-3 -(dimethylamino)- 1 - ((phenylsulfanyl)methyl)propyl)amino)-3-nitrobenzenesulfonamide) (ABT-737), N-(4-(4-((2- (4-chlorophenyl)-5 ,5 -dimethyl- 1 -cyclohex- 1 -en- 1 -yl)methyl)piperazin- 1 -yl)benzoyl)-4- ((( 1
  • Bcr-Abl kinase inhibitors include DASATINIB ® (BMS-354825), GLEEVEC ® (imatinib) and the like.
  • CDK inhibitors include AZD-5438, BMI-1040, BMS-032, BMS-387, CVT-2584, flavopyridol, GPC-286199, MCS-5A, PD0332991, PHA-690509, seliciclib (CYC-
  • COX-2 inhibitors include ABT-963, ARCOXIA ® (etoricoxib), BEXTRA ® (valdecoxib), BMS347070, CELEBREX ® (celecoxib), COX- 189 (lumiracoxib), CT-3, DERAMAXX ® (deracoxib), JTE-522, 4-methyl-2-(3 ,4-dimethylphenyl)- 1 -(4- sulfamoylphenyl-1H-pyrrole), MK-663 (etoricoxib), NS-398, parecoxib, RS-57067, SC-58125, SD-8381, SVT-2016, S-2474, T-614, VIOXX ® (rofecoxib) and the like.
  • EGFR inhibitors include ABX-EGF, anti-EGFR immunoliposomes, EGF-vaccine, EMD-7200, ERBITUX ® (cetuximab), HR3, IgA antibodies, IRESSA ® (gefitinib),
  • TARCEVA ® (erlotinib or OSI-774), TP-38, EGFR fusion protein, TYKERB ® (lapatinib) and the like.
  • ErbB2 receptor inhibitors include CP-724-714, CI-1033 (canertinib), HERCEPTIN ® (trastuzumab), TYKERB ® (lapatinib), OMNITARG ® (2C4, petuzumab), TAK- 165 , GW-572016 (ionafarnib), GW-282974, EKB-569, PI- 166, dHER2 (HER2 vaccine), APC-8024 (HER-2 vaccine), anti-HER/2neu bispecific antibody, B7.her2IgG3, AS HER2 trifunctional bispecfic antibodies, mAB AR-209, mAB 2B- 1 and the like.
  • Histone deacetylase inhibitors include depsipeptide, LAQ-824, MS-275, trapoxin, suberoylanilide hydroxamic acid (SAHA), TSA, valproic acid and the like.
  • HSP-90 inhibitors include 17-AAG-nab, 17-AAG, CNF-101, CNF-1010, CNF-2024, 17-DMAG, geldanamycin, IPI-504, KOS-953, MYCOGRAB ® (human recombinant antibody to HSP-90), NCS-683664, PU24FC1, PU-3, radicicol, SNX-2112, STA-9090 VER49009 and the like.
  • Inhibitors of apoptosis proteins include ApoMab (a fully human affinity-matured IgGl monoclonal antibody), antibodies that target TRAIL or death receptors (e.g., pro- apoptotic receptor agonists DR4 and DR5), conatumumab, ETR2-ST01, GDCO 145, (lexatumumab), HGS-1029, LBY-135, PRO-1762 and tratuzumab.
  • ApoMab a fully human affinity-matured IgGl monoclonal antibody
  • antibodies that target TRAIL or death receptors e.g., pro- apoptotic receptor agonists DR4 and DR5
  • conatumumab e.g., ETR2-ST01, GDCO 145, (lexatumumab), HGS-1029, LBY-135, PRO-1762 and tratuzumab.
  • MEK inhibitors include ARRY-142886, ARRY-438162 PD-325901, PD-98059 and the like.
  • mTOR inhibitors include AP-23573, CCI-779, everolimus, RAD-OOl, rapamycin, temsirolimus and the like.
  • Non-steroidal anti-inflammatory drugs include AMIGESIC (salsalate), DOLOBID (difiunisal), MOTRIN ® (ibuprofen), ORUDIS ® (ketoprofen), RELAFEN ® (nabumetone),
  • FELDENE piroxicam
  • ibuprofen cream ALEVE
  • NAPROSYN naproxen
  • VOLTAREN ® diclofenac
  • INDOCIN ® indomethacin
  • CLINORIL ® sinalindac
  • TOLECTIN ® tolmetin
  • LODINE ® etodolac
  • TORADOL ® ketorolac
  • DAYPRO ® oxaprozin
  • PDGFR inhibitors include C-451, CP-673, CP-868596 and the like.
  • Platinum chemotherapeutics include cisplatin, ELOXATIN (oxaliplatin) eptaplatin,
  • Polo-like kinase inhibitors include BI-2536 and the like.
  • Thrombospondin analogs include ABT-510, ABT-567, TSP-I and the like.
  • VEGFR inhibitors include AVASTIN ® (bevacizumab), ABT-869, AEE-788, ANGIOZYMETM (a ribozyme that inhibits angiogenesis (Ribozyme Pharmaceuticals (Boulder, CO.) and Chiron, (Emeryville, CA)) , axitinib (AG- 13736), AZD-2171, CP-547,632, IM-862, MACUGEN (pegaptamib), NEXAVAR ® (sorafenib, BAY43-9006), pazopanib (GW-786034), vatalanib (PTK-787, ZK-222584), SUTENT ® (sunitinib, SU- 11248), VEGF trap, ZACTIMATM (vandetanib, ZD-6474) and the like.
  • AVASTIN ® bevacizumab
  • ABT-869 ABT-869
  • AEE-788 ANGIOZY
  • Antibiotics include intercalating antibiotics aclarubicin, actinomycin D, amrubicin, annamycin, adriamycin, BLENOXANE (bleomycin), daunorubicin, CAELYX or
  • MYOCET liposomal doxorubicin
  • elsamitrucin epirbucin
  • glarbuicin ZAVEDOS
  • rebeccamycin stimalamer, streptozocin, VALSTAR (valrubicin), zinostatin and the like.
  • Topoisomerase inhibitors include aclarubicin, 9-aminocamptothecin, amonafide, amsacrine, becatecarin, belotecan, BN-80915, CAMPTOSAR (irinotecan hydrochloride),
  • PHARMORUBICIN epirubicin
  • etoposide exatecan
  • 10-hydroxycamptothecin gimatecan
  • lurtotecan mitoxantrone
  • orathecin pirarbucin
  • pixantrone rubitecan
  • sobuzoxane SN-38
  • tafluposide topotecan and the like.
  • Antibodies include AVASTIN (bevacizumab), CD40-specif ⁇ c antibodies, chTNT- 1/B, denosumab, ERBITUX ® (cetuximab), HUMAX-CD4 ® (zanolimumab), IGFlR-specif ⁇ c antibodies, lintuzumab, PANOREX ® (edrecolomab), RENCAREX ® (WX G250),
  • Hormonal therapies include ARIMIDEX ® (anastrozole), AROMASIN ® (exemestane), arzoxifene, CASODEX ® (bicalutamide), CETROTIDE ® (cetrorelix), degarelix, deslorelin, DESOPAN (trilostane), dexamethasone, DROGENIL , (flutamide), EVISTA ® (raloxifene), AFEMATM (fadrozole), FARESTON ® (toremifene), FASLODEX ®
  • Deltoids and retinoids include seocalcitol (EB 1089, CB 1093), lexacalcitrol (KH 1060), fenretinide, PANRETIN ® (aliretinoin), ATRAGEN ® (liposomal tretinoin), TARGRETIN ® (bexarotene), LGD- 1550 and the like.
  • PARP inhibitors include ABT-888, olaparib, KU-59436, AZD-2281, AG-014699,
  • Plant alkaloids include, but are not limited to, vincristine, vinblastine, vindesine, vinorelbine and the like.
  • Proteasome inhibitors include VELC ADE ® (bortezomib), MG 132, NPI-0052, PR-171 and the like.
  • immunologicals include interferons and other immune-enhancing agents.
  • Interferons include interferon alpha, interferon alpha-2a, interferon alpha-2b, interferon beta, interferon gamma- Ia, ACTIMMUNE (interferon gamma- Ib), or interferon gamma-nl, combinations thereof and the like.
  • Other agents include ALF AFERONE ,(IFN- ⁇ ), BAM- 002 (oxidized glutathione), BEROMUN ® (tasonermin), BEXXAR ® (tositumomab),
  • CAMPATH (alemtuzumab), CTLA4 (cytotoxic lymphocyte antigen 4), decarbazine,
  • SSM Session Management
  • WF-IO Tetrachlorodecaoxide
  • TCDO Tetrachlorodecaoxide
  • PROLEUKIN ® aldesleukin
  • ZADAXIN ® thymalfasin
  • ZENAP AX ® diaclizumab
  • ZEVALIN ® 90Y-Ibritumomab tiuxetan
  • Biological response modifiers are agents that modify defense mechanisms of living organisms or biological responses, such as survival, growth, or differentiation of tissue cells
  • 59 to direct them to have anti-tumor activity include include krestin, lentinan, sizof ⁇ ran, picibanil PF-3512676 (CpG-8954), ubenimex and the like.
  • Pyrimidine analogs include cytarabine (ara C or Arabinoside C), cytosine arabinoside, doxifluridine, FLUDARA (fludarabine), 5-FU (5-fluorouracil), floxuridine, GEMZAR
  • Purine analogs include LANVIS ® (thioguanine) and PURI-NETHOL ® (mercaptopurine) .
  • Antimitotic agents include batabulin, epothilone D (KOS-862), N-(2-((4- hydroxyphenyl)amino)pyridin-3 -yl)-4-methoxybenzenesulfonamide, ixabepilone (BMS 247550), paclitaxel, TAXOTERE ® (docetaxel), PNU100940 (109881), patupilone, XRP-9881 (larotaxel), vinflunine, ZK-EPO (synthetic epothilone) and the like.
  • Radiosensitizeser that enhance the efficacy of radiotherapy.
  • radiotherapy include external beam radiotherapy, teletherapy, brachtherapy and sealed, unsealed source radiotherapy and the like.
  • compounds having Formula I may be combined with other chemptherapeutic agents such as ABRAXANETM (ABI-007), ABT- 100 (farnesyl transferase inhibitor), ADVEXIN ® (Ad5CMV-p53 vaccine), ALTOCOR ® or MEVACOR ® (lovastatin), AMPLIGEN ® (poly Lpoly C12U, a synthetic RNA), APTOS YN ® (exisulind), AREDIA ® (pamidronic acid), arglabin, L-asparaginase, atamestane (l-methyl-3,17-dione-androsta-1,4- diene), AVAGE (tazarotene), AVE-8062 (combreastatin derivative) BEC2 (mitumomab), cachectin or cachexin (tumor necrosis factor), canvaxin (vaccine), CEAVAC (cancer vaccine), CELEUK ® (celmoleukin), CEPLENE
  • TransMID-107RTM diphtheria toxins
  • dacarbazine dactinomycin, 5,6- dimethylxanthenone-4-acetic acid (DMXAA), eniluracil, EVIZONTM (squalamine lactate), DIMERICINE ® (T4N5 liposome lotion), discodermolide, DX-8951f (exatecan mesylate), enzastaurin, EPO906 (epithilone B), GARDASIL (quadrivalent human papillomavirus (Types 6, 11, 16, 18) recombinant vaccine), GASTRIMMUNE ® , GENASENSE ® , GMK
  • PANVAC -VF investigational cancer vaccine
  • pegaspargase PEG Interferon A
  • phenoxodiol procarbazine
  • rebimastat REMOVAB (catumaxomab)
  • REVLIMID lenalidomide
  • RSRl 3 efaproxiral
  • SOMATULINE ® LA lanreotide
  • CaBLES comprise one or more non-cationic lipids, one or more cationic lipids having Formula I and one or more polyethylene glycol (PEG)-lipid conjugate.
  • PEG polyethylene glycol
  • Lipid-Based Particles of the present invention are defined as CaBLES which further comprise one or more therapeutic agent(s). These particles have mean diameter sizes of 50- 300 nm, of which 50-250 nm is preferred and 50-200 nm is most preferred.
  • Functional CaBLES effectively encapsulate nucleic acids, (e.g., single stranded or double stranded DNA, single stranded or double stranced RNA, RNAi, siRNA, and the like). Suitable nucleic acids include, but are not limited to, plasmids, antisense oligonucleotides, ribozymes as well as other poly- and oligonucleotides.
  • the nucleic acid encodes a product, e.g., a therapeutic product, of interest.
  • the CaBLES of the present invention can be used to deliver the nucleic acid to a cell (e.g., a cell in a mammal) for, e.g., expression of the nucleic acid or for silencing of a target sequence expressed by the cell.
  • the nucleic acid is a siRNA molecule that silences the gene of interest, with efficiencies from about 50-100%, and more preferably between about 80-100%.
  • the therapeutic agents that can be delivered with CaBLES include RNA, antisense oligonucleotide, a DNA, a plasmid, a ribosomal RNA (rRNA), a micro RNA (miRNA), transfer RNA (tRNA), a small inhibitory RNA (siRNA), small nuclear RNA (snRNA), chimeric nucleic acids, an antigen, fragments thereof, a protein, a peptide, small-molecules, or mixtures thereof.
  • RNA' s such as small inhibitory RNA or microRNA.
  • the siRNA can have varying lengths (10-200 bps) and structures (hairpins, single/double strands, bulges, nicks/gaps, mismatches) and processed in the cell to provide active gene silencing.
  • a double- stranded siRNA can have the same number of nucleotides on each strand (blunt ends) or asymmetric ends (overhangs). The overhang of 1-2 nucleotides can be present on the sense and/or the antisense strand, as well as present on the 5'- and/ or the 3 '-ends of a given strand.
  • Suitable siRNA sequences can be identified using means known in the art (e.g., methods described in Elbashir, et al, Nature 411 :494-498 (2001) and Elbashir, et al, EMBO
  • Non-cationic lipids have a neutral charge or an anionic charge at physiological pH.
  • a neutral lipid also known as a "helper lipid,” has no net charge at physiological pH.
  • helper lipid has no net charge at physiological pH.
  • These lipids can also be zwitterionic.
  • Polyethylene glycol (PEG)-lipid conjugates are used to minimize particle aggregation in solution, provide increased in vivo serum circulation, and enhance distribution of nanoparticles to organs, tissues, cell types, and tumors of interest.
  • These shielding lipids consist of a lipid portion linked to a "PEG” portion via carbamate, ester, amide, ether, amine, thioether, or dithiol linkages.
  • PEG is a polyethylene glycol consisting of repeating C2H4O units with an average molecular weight between 500 to 10,000 daltons and may be substituted by alkoxy, acyl, alkyl, or aryl.
  • the PEG can be substituted at its terminus with one or more of the following functional groups: hydroxy, methoxy, primary, secondary, or tertiary amine, thiol, thioether, thiopyridyl, dithiol, maleimide, or ester.
  • the CaBLES and/or Lipid Based Particles may target using targeting moieties that are specific to a cell type or tissue.
  • targeting moieties such as ligands, cell surface receptors, glycoproteins, vitamins, (e.g., ribolflavin) and moncoleonal antibodies, has been previously described (see, e.g., U.S. Pat. Nos. 4,957,773 and 4,603,044).
  • the targeting moeities can comprise the entire entire protein or fragments thereof.
  • the targeting moiety is a small protein, or peptide.
  • the targeting moiety is a small-molecule.
  • Cationic lipids are those having one or more moieties that are positively charged at a physiologically relevant pH, typically between 4-8. Particular cationic lipids are as shown in Formula I. Examples of cationic lipids that are useful for the practice of this invention include, but are not limited to, N,N-dioleyl-N,N-dimethylammonium chloride, DC-Choi, 1,3-dioleoyloxy-2-(6-carboxyspermyl)-propyl amide, dioctadecylamidoglycyl spermine, N,N-distearyl-N,N-dimethylammonium bromide, N-(2,3-dioleyloxy)propyl)-N,N- dimethylammonium chloride, 1,2-dioleoyl-3-trimethylammonium-propane chloride, 1,2-dilineoyl-3-dimethylammonium-propane, N-(l-(
  • Lipid-Based Particles are a mixture of one or more cationic lipids of Formula (I), one or more non-cationic lipids, one or more PEG-lipid conjugates, and one or more therapeutic agents.
  • Specific Lipid-Based Particles comprise the following lipid mixtures: cationic lipid(s) (about 2-60% by weight), non-cationic lipid(s) (about 5-90% by weight), and PEG-lipid conjugate(s) (about 0.1-20%).
  • Non-cationic 10 0.5 lipid
  • the mixing solution of cationic lipids, cholesterol, non-cationic lipids and PEG-lipids was prepared in ethanol (total concentration at 10 mg/mL).
  • siSTABLE purchased from ThermoFisher
  • % stock solution was prepared in 10 mg/mL of solution by dissolving 10 mg siRNA in 1 mL of RNAse-free UltraPure Water.
  • the calculated amount of siRNA solution was added to 1 mL of citrate buffer (pH 4.0, 20 mM), to provide an siRNA concentration of 0.2 mg/mL, and warmed to 6OC.
  • the calculated amount of lipid solution was warmed to 6OC, transferred to a 0.5 mL syringe with 28/4 gauge needle, and injected into the citrate buffer with stirring at 6OC.
  • 3 mL of PBS solution at room temperature (pH 7.4) was added into the lipid mixture with stirring.
  • the Lipid-Based Particle solution was cooled to room temperature.
  • siRNA concentrations were measured using Quanti-iT RiboGreen RNA reagent (Molecular Probes, (Rl 1490)). Vesicle sizes were characterized by dynamic light scattering with a DynaProTM Plate Reader (Wyatt Technology) in 96-well half-area UV plate (Coring) after diluting the formulation sample (20 ⁇ L) in phosphate buffered saline (80 ⁇ L) at a pH of about 7-8. A 1% agarose gel-based assay was used for analyzing nuclease degradation and protection. Encapsulation efficiency (EE) was calculated using data obtained from a RiboGreen assay.
  • EE Encapsulation efficiency
  • RNA concentration and encapsulation efficiency were determined using a Quant-iT ® Ribogreen RNA reagent and kit available from Invitrogen.
  • the siRNA was released from the Lipid-Based Particle using one of the following reagents: ethanol, Triton X-100, or
  • the siRNA concentration is quantified using fluorescent reading at 480 nm/520 nm.
  • Particle Sizing Assay Particle sizes and size distributions were characterized by using dynamic light scattering (DLS).
  • DLS plate reader (DynaproTM, Wyatt Technology) was used for the DLS measurement. This DLS plate reader uses an 830 nm laser and the scattering angle is 158°. It also can control temperature from 4°C to 70°C.
  • a 96-well format was employed for the samples. Samples for DLS analysis were prepared by mixing 20 ⁇ L of each sample stock solution with 80 ⁇ L PBS directly in the 96-well plate (#3697, Corning). Sample mixing was accomplished using a microplate shaker (Orbis, Mikura Ltd.).
  • the lipid solution was prepared (10 mg/ml) by dissolving the lipid in 200 proof ethanol.
  • the lipid mixture solution is prepared according to the above composition in Table
  • siRNA (TetR_ODC_12, G.G.G.G.A.A.A.G.C.U.G.G.C.A.A.G.A.U.U.U) (ThermoFisher) solution is prepared in a concentration of 10 mg/ml by dissolving 10 mg siRNA in 1 ml of DNAse/RNAse-free distilled water.
  • a round bottom flask was submerged into a 65°C water bath.
  • Citrate buffer (37.5 ml) of pH 4.0 was pipetted into the flask. The solution was stirred by a magnetic stirring bar at a speed of 900 rpm. Both the pH 4.0 citrate buffer and the lipid solution were pre-warmed in the 65°C water bath for about 3 minutes.
  • a siRNA solution (0.5 ml) was pipetted into the pH 4.0 citrate buffer. The 12.5 ml lipid mixture solution was injected through a 27 gauge needle into the citrate buffer in about 30 seconds. The needle tip was inserted into the solution during the injection. The resulting solution was stirred for 5 minutes at a speed of 900 rpm.
  • the flask was pulled up from the water bath and a 50 ml pH 7.4 PBS buffer was added into the flask. The final solution was further mixed at a speed of 900 rpm for 5 minutes.
  • a dialysis filter (Millipore, 10OK, Cat# PXB100C50) was used to remove ethanol in the above solution.
  • 20 ml of pH 7.4 PBS was added to the sample solution.
  • the diafiltration was continued until the volume was reduced to 20 ml.
  • the diafiltration process was repeated 4 times.
  • the volume of the sample solution was reduced to about 12 ml and pH 7.4 PBS was added to make the final volume of 15 ml.
  • the 15 ml solution was filtered sequentially through the 0.45 and 0.22 ⁇ m sterile PVDF membrane filters (Millipore) and immediately transferred into a sterile vial.
  • lipid-based particles were prepared as described above.
  • the particle solution 60 ⁇ L was pipetted into a disposable cuvette (UVette, Eppendorf, cat# 952010051) and measured in the "General Purpose" mode. Attenuator and position were optimized by the device. Measurements were performed using a Zetasizer Nano ZS (Malvern Instruments) equipped with a 4 mW He-Ne laser at a wavelength of 633 nm at 25 °C. Scattered light was detected at a 173° backward scattering angle. The viscosity and refractive index of water at 25 °C was used for data analysis with the Dispersion Technology Software 5.00 (Malvern Instruments).
  • MDA435-TetR-Luc cells (The positive readout reporter cell line MDA435-TetR-Luc contained a stably integrated copy of the luciferase gene expressed from a CMV promoter containing the tetR operator site. In addition, gene coding for a destabilized TetR protein was expressed in this cell line.) were plated in 96 well plate at a density of 1OK cells per well in 100 ul of DMEM (Dulbecco's Modified Eagles Medium, Invitrogen Corp.) containing 10% fetal bovine serum (Invitrogen Corp.).
  • DMEM Dulbecco's Modified Eagles Medium, Invitrogen Corp.
  • 10% fetal bovine serum Invitrogen Corp.
  • Lipid-Based Particles were made in DMEM+10% fetal bovine serum medium, 10 ul of the diluted material was transferred into each well in triplicate. Transfected cells were further incubated at 37°C for a period of 72 hours. Supernatent from each well was removed and cells were assayed for luciferase activity (Steady GIo kit , ProMega Corp.) as per the manufacturers recommendation. Positive controls included cells treated with 100 ul of doxycycline at 0.5 mg/ml, 20 nM tetR siRNA transfected with lipofectamine (Invitrogen Corp.) or untreated
  • the graphs represent average of triplicate readings of the Lipid-Based Particles treated sample divided by the average of readings from 9 wells treated with doxycycline.
  • Formulated or unformulated siRNAs were administrated via tail vein (i.v) injection.
  • DAB (3,3'-diaminobenzidine) was used as the chromogen.
  • 1HC images were acquired using the Nikon TE2000 inverted microscope.
  • the ⁇ -galactosidase staining was evaluated by 2 people independently based on the scoring system listed below. The average of the score was calculated for each tumor.
  • IHC scoring system - there was no staining, a small area of weak staining, or disperse strong single cell staining
  • Bioluminescence Imaging and Analysis In vivo bioluminescence imaging and analysis were conducted on the IVIS 200 system using the Living Image acquisition and analysis software (Caliper Life Science,
  • luciferin Promege, Madison, WI
  • mice were anesthetized with isofluorane.
  • ROI Regions of interest
  • cationic lipid transfection agents that typically contain either a dimethylamino- or trimethylammonium cationic head group (e.g., DOTAP, DODAP, and DOTMA (Avanti Polar Lipids Inc., Alabaster, AL).
  • DOTAP dimethylamino- or trimethylammonium cationic head group
  • DODAP DODAP
  • DOTMA Advanti Polar Lipids Inc., Alabaster, AL
  • changing the head group to the related diethylamino cationic head group may diminish activity (vide infra, Example 57).
  • a formulation that employs a cationic lipid having a heterocyclic amine, such as pyrrolidine unexpectedly provided in vitro transfection activity as well as efficient in vivo delivery.
  • the in vitro transfection efficiency of a given formulation may or may not predict for in vivo delivery.
  • the in vivo delivery may depend upon the properties of other co-lipid components in the formulation. Properties of the co-lipids that may modulate in vivo delivery, include for example, PEG lipid alkyl length, PEG polymer length, concentration of the PEG lipid conjugate, presence and concentration of neutral helper lipid, as well as the manner of which the co-lipid components are formulated (Sadzuka, et.al., J. Liposome Research, 13,2, (2003) 157-172; Sadzuka, et. al, Int. J.
  • the aggregate effect of these co-lipids and their formulation impacts a set of parameters that includes for example particle stabilization, serum stability, circulation half- life, particle internalization, intracellular release of the therapeutic agent. These factors in total are likely to mitigate effective in vivo delivery.
  • ADDP means 1,1'- (azodicarbonyl)dipiperidine
  • AD-mix- ⁇ means a mixture Of (DHQD) 2 PHAL, K 3 Fe(CN) 6 , K 2 CO 3 and K 2 SO 4 );
  • AIBN means 2,2'-azobis(2-methylpropionitrile);
  • 9-BBN means 9- borabicyclo(3.3.1)nonane;
  • Cp means cyclopentadiene;
  • DHQD) 2 PHAL means hydroquinidine 1 ,4-phthalazinediyl diethyl ether;
  • DBU means 1,8-diazabicyclo(5.4.0)undec- 7-ene;
  • DCC means dicyclohexylcarbodiimide;
  • DIBAL means diisobutylaluminum hydride;
  • DIEA means diisopropylethylamine
  • DMAP means N,N-dimethylaminopyridine
  • DME means 1,2-dimethoxyethane
  • DMF means N,N-dimethylformamide
  • dmpe means 1,2- bis(dimethylphosphino)ethane
  • DMSO means dimethylsulfoxide
  • dppa means diphenylphosphoryl azide
  • dppb means 1,4-bis(diphenylphosphino)butane
  • dppe means 1,2- bis(diphenylphosphino)ethane
  • dppf means l,r-bis(diphenylphosphino)ferrocene
  • dppm means l,l-bis(diphenylphosphino)methane
  • EDAC means l-(3-dimethylaminopropyl)-3- ethylcarbodiimide
  • Fmoc means fluorenylmeth
  • the diol of Formula (5) may commercially available or synthesized as described in Example 9 and reacted with a compound of Formula (3) to obtain a compound of Formula (1).
  • the reaction typically requires the use of a base such as but not limited to sodium hydride and a solvent such as but not limited to toluene at elevated temperatures.
  • R 3 and R 4 can be the same, two equivalents of (3) can be used. IfR 3 and R 4 are to be different, one equivalent of (3) can be used to obtain a compound wherein R 4 is H after purification. This intermediate can then be reacted with CH 3 (SOs)R 4 to obtain a compound of Formula (I).
  • Compounds of Formula (I), wherein R 1 and R 2 when taken together are heteroaryl or heterocycloalkyl, can be prepared by reacting compounds of Formula (5) and base, such as but not limited to sodium hydride, with a compound of Formula (3).
  • the reaction is typically performed in at an elevated temperature in a solvent such as but not limited to toluene. If it is desired for R 3 and R 4 to be the same, two equivalents of (3) can be used. IfR 3 and R 4 are to be different, one equivalent of (3) can be used to obtain a compound wherein R 4 is H after purification. This intermediate can then be reacted with CHs(SOs)R 4 to obtain a compound of Formula (I).
  • the diol of Formula (5) which is commercially available or synthesized as described in Example 9, can be reacted with a compound of Formula (6) to obtain a compound of Formula (I), as shown in Scheme 4.
  • the reaction typically requires the use solvent such as but not limited to dichloromethane in the presence of DMAP and EDC.
  • R 3 and R 4 are to be the same, two equivalents of (6) can be used. IfR 3 and R 4 are to be different, one equivalent of (6) can be used to obtain a compound wherein R 4 is H after purification. This intermediate can then be reacted with R 4 C(O)OH to obtain a compound of Formula (I).
  • a bromide of Formula (7) which may be commercially available, can be reacted with magnesium in diethyl ether to obtain the compound of Formula (8).
  • a compound of Formula (6) can be reacted with oxalyl chloride in CHCI3 followed by the addition of a compound of Formula (8) to obtain a compound of Formula (9).
  • the diol of Formula (5) which may be commercially available or synthesized as described in Example 9, can be reacted with a compound of Formula (9), which may be commercially available, to obtain a compound of Formula (I).
  • the reaction typically requires the use solvent such as but not limited to toluene at elevated temperatures in the presence of an acid catalyst.
  • EXAMPLE 4 l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)-4-methyl-1,4-diazepane Prepared as described in EXAMPLE 3 by substituting 1 -methyl- 1 ,4-diazepane for 1- methyl-piperazine.
  • Linoleyl methane sulfonate (1 g) was added to a mixture of 3-(4-phenylpiperazin-1- yl)propane-1,2-diol (241 mg) and 60% oily sodium hydride (200 mg) in toluene (5 mL). After 5 minutes, the mixture was heated at 100°C in a sealed vial for 2 hours, cooled to room temperature, quenched with methanol and treated with ethyl acetate and water. The extract was washed with brine and dried over Na 2 SO 4 , filtered and concentrated. The concentrate was purified by flash chromatography on silica gel (0-15% methanol in dichloromethane). MS (ESI-) m/e 734.6 (M+H) + ; 1 H NMR (400 MHz, CDCl 3 ) ⁇ 7.16-7.34 (m, 2H), 6.75-7.02
  • EXAMPLE 7 l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)piperidine
  • 3-(piperidin-1-yl)propane-1,2-diol (0.2 g) and NaH (0.32 g) in toluene (12.56 ml) was stirred at room temperature for 30 minutes, treated with (9Z,12Z)-octadeca- 9,12-dienyl methanesulfonate (0.952 g) in toluene (1 mL), heated at 9O°C for 4 hours, cooled to room temperature, quenched with methanol and treated with ethyl acetate and water.
  • EXAMPLE 10 l-((2S)-2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidine Prepared as described in EXAMPLE 9 by substituting (R)-3-chloropropane-1,2-diol for (S)-3-chloropropane-1,2-diol.
  • EXAMPLE 11 l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)-4-ethylpiperazine
  • 1-ethylpiperazine (625 mg) and 3-chloropropane-1,2-diol (528 mg) in dioxane (4 mL) was heated in a microwave (Biotage Initiator) at 14O°C for 40 minutes, cooled, added to saturated sodium bicarbonate solution and washed with chloroform.
  • the extract was dried over anhydrous Na 2 SO 4 , filtered and concentrated.
  • the concentrate was dissolved in toluene (5 mL) and treated with inoleyl methane sulfonate (1 g) and 60% oily sodium hydride (250 mg). After 5 minutes at room temperature, the mixture was heated at 100°C in a sealed vial for 2 hours. After cooling to room temperature, the mixture was
  • EXAMPLE 14 l-((2S)-2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)-4-methylpiperazine Prepared as described in EXAMPLE 11 by substituting 1-methylpiperazine for 1- ethylpiperazine and (R)-3-chloropropane-1,2-diol for 3-chloropropane-1,2-diol.
  • EXAMPLE 24 was prepared following the procedure detailed for EXAMPLE 1 in which linoleyl methane sulfonate was replaced with (5Z,8Z,1 lZ,14Z)-icosa-5, 8,11,14- tetraenyl methane sulfonate. MS (ESI(+)) m/e 691 (M+H) + .
  • EXAMPLE 25 was prepared by taking EXAMPLE 22A and treating it with (5Z,8Z,1 lZ,14Z)-icosa-5,8,l 1,14-tetraenyl methane sulfonate. MS (ESI(+)) m/e 667 (M+H) + .
  • EXAMPLE 27 l- ⁇ 2-[(9E, 12E)-octadeca-9,12-dienyloxy]-3-[(9Z,12Z)-octadeca-9,12- dienyloxy]propyl ⁇ pyrrolidine
  • NaH 0.046 g, 1.94 mmol
  • 9E,12E)- octadeca-9,12-dienyl methanesulfonate 0.5 g, 1.45 mmol
  • EXAMPLE 28 l-[2,3-bis(tetradecyloxy)propyl]pyrrolidine EXAMPLE 28 was prepared following the procedure detailed for EXAMPLE 1 in which linoleyl methane sulfonate was replaced with tetradecyl methanesulfonate. MS (ESI(+)) m/e 539 (M+H) + .
  • EXAMPLE 29 was prepared following the procedure detailed for EXAMPLE 1 in which linoleyl methane sulfonate was replaced with octadecyl methanesulfonate.
  • EXAMPLE 30 was prepared following the procedure detailed for EXAMPLE 1 in which linoleyl methane sulfonate was replaced with oleyl methane sulfonate. MS (ESI(+)) m/e 651 (M+H) + .
  • EXAMPLE 32E l-(2,3-bis((9Z,12Z)-octadeca-9,12-dienyloxy)propyl)pyrrolidin-3-ol
  • pyrrolidin-3-ol 0.14 g, 1.64 mmol
  • acetic acid 0.25 g, 4.1 mmol
  • the silica was loaded into an Analogix DASI module, and the product was isolated by flash chromatography (Analogix, SF25x60g, 1% methanol/ethyl acetate for six column volumes, then 1-10% methanol/ethyl acetate over six column volumes) to give the title compound.
  • MS (ESI(+)) m/e 607 (M+H) + .
  • EXAMPLE 33 was prepared as described in EXAMPLE 27, replacing (9E,12E)- octadeca-9,12-dienyl methanesulfonate with (Z)-octadec-9-enyl methanesulfonate.
  • the aqueous layer was extracted twice with ethyl acetate and the extract was dried (Na 2 SO 4 ), filtered, and concentrated.
  • the concentrate was purified by flash column chromatography (Analogix hexanes: ethyl acetate, 0-75%) to afford the title compound.
  • EXAMPLE 35B was added to tetrahydrofuran (20 rnL) and 2N HCl (20 mL), and the mixture was stirred at room temperature for 30 minutes. 5N NaOH was added until the solution was basic, and the aqueous layer was extracted with chloroform. The extract was dried (MgSO 4 ), filtered and concentrated by rotary evaporation and the concentrate was used in the next step without further purification. MS (ESI) m/z 285.9 (M+H) + .
  • EXAMPLE 35D 500 mg was added to methanol/dichloromethane/ethyl acetate (1/1/1, 10 niL) and combined with catalytic Pd/C (10%). Hydrogen was introduced via a balloon, and the mixture was stirred overnight then filtered through Celite®. The filtrate was concentrated and the concentrate was used in the next step without further purification.
  • EXAMPLE 36B was prepared using the procedure described for EXAMPLE 35E, substituting EXAMPLE 36A for EXAMPLE 35D.
  • MS (ESI) m/z 498.5 (M+H) + ; 1 H NMR (300 MHz, CDCl 3 ) ⁇ ppm 8.24 (s, 2H) 3.53-3.70 (m, 1H) 3.34-3.53 (m, 6H) 3.07-3.34 (m, 2H) 1.87-2.13 (m, 2H) 1.48-1.67 (m, 4H) 1.16-1.39 (m, 44H) 0.82-0.94 (m, 6H).
  • EXAMPLE 36C was prepared using the procedure described for EXAMPLE 35F, substituting EXAMPLE 26B for EXAMPLE 35E.
  • MS (MALDI) m/z 2617.6; 1 H NMR (300 MHz, CDCl 3 ) ⁇ 3.95-4.02 (m, 2H) 3.83-3.92 (m, 1H) 3.68-3.72 (m, 1H) 3.65 (m, 180H) 3.35- 3.60 (m, 10H) 1.59-1.73 (m, 2H) 1.49-1.60 (m, 4H) 1.18-1.36 (m, 44H) 0.82-0.94 (m, 6H).
  • EXAMPLE 37A was prepared using the procedure described for EXAMPLE 36 A, substituting hexadecyl methanesulfonate for tetradecyl methanesulfonate.
  • MS (ESI) m/z 734.6 (M+H) + ; 1 H NMR (300 MHz, CDCl 3 ) ⁇ ppm 7.15-7.41 (m, 10H) 3.12-3.64 (m, HH) 2.41-2.64 (m, 2H) 1.35-1.80 (m, 6H) 1.15-1.34 (m, 52H) 0.81-0.94 (m, 6H).
  • EXAMPLE 37B was prepared using the procedure described for EXAMPLE 36B, substituting EXAMPLE 37A for EXAMPLE 36A.
  • MS (ESI) m/z 554.6 (M+H) + ; 1 U NMR (300 MHz, CDCl 3 ) ⁇ ppm 8.12-8.38 (m, 2H) 3.54-3.70 (m, 1H) 3.33-3.53 (m, 6H) 3.06-3.33 (m, 2H) 1.84-2.14 (m, 2H) 1.46-1.71 (m, 4H) 1.14-1.37 (m, 52H) 0.81-0.94 (m, 6H).
  • EXAMPLE 37C was prepared using the procedure described for EXAMPLE 35F, substituting EXAMPLE 37B for EXAMPLE 35E.
  • MS (MALDI) m/z 2866.7; 1 U NMR (300 MHz, CDCl 3 ) ⁇ ppm 3.98 (s, 2H) 3.84-3.91 (m, 1H) 3.60-3.68 (m, 180H) 3.36-3.60 (m, HH) 1.50-1.72 (m, 6H) 1.26 (s, 52H) 0.84-0.92 (m, 6H).
  • EXAMPLE 38A N,N-dibenzyl-3 ,4-bis(octadecyloxy)butan- 1 -amine
  • EXAMPLE 558 A was prepared using the same procedure described for EXAMPLE 36A, substituting octadecyl methanesulfonate for tetradecyl methanesulfonate.
  • EXAMPLE 38B was prepared using the same procedure described for EXAMPLE 35E, substituting EXAMPLE 38A for EXAMPLE 35D.
  • LCMS APCI
  • EXAMPLE 38C was prepared using the same procedure described for EXAMPLE 35F, substituting EXAMPLE 38B for EXAMPLE 35E.
  • MS (MALDI) m/z 2773.6; 1 H NMR (300 MHz, CDCl 3 ) ⁇ ppm 3.95-4.01 (m, 2H) 3.84-3.91 (m, 1H) 3.59-3.70 (m, 180H) 3.27- 3.59 (m, HH) 1.49-1.86 (m, 6H) 1.18-1.35 (m, 60H) 0.80-0.94 (m, 6H).
  • EXAMPLE 40 3,6,9,12,15, 18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72,75,78,81,84,87,90,93,
  • EXAMPLE 40 was prepared using the same procedure described for EXAMPLE 39, substituting EXAMPLE 37B for EXAMPLE 36B.
  • EXAMPLE 41 was prepared using the same procedure described for EXAMPLE 39, substituting EXAMPLE 38B for EXAMPLE 36B.
  • EXAMPLE 42 was prepared using the same procedure described for EXAMPLE 35F, substituting RAPP 12 2000-35 (Rapp Polymere) for mPEG2000-SCM.
  • MS (MALDI) m/z 2584.3; 1 H NMR (300 MHz, CDCl 3 ) ⁇ ppm 6.43-6.61 (m, 2H) 3.60-3.68 (m, 200H) 3.36-3.58 (m, 16H) 2.42-2.57 (m, 4H) 1.49-1.85 (m, 6H) 1.19-1.35 (m, 52H) 0.82-0.92 (m, 6H).
  • EXAMPLE 43 was prepared using the same procedure described for EXAMPLE 35F, substituting mPEG-NPC (Creative PEGWorks) for mPEG2000-SCM (Laysan Bio, Inc.). MS (MALDI) m/z 2588.5; 1 H NMR (300 MHz, CDCl 3 ) ⁇ ppm 5.14 (m, 1H) 4.17-4.26 (m, 3H) 4.01-4.11 (m, 1H) 3.83-3.91 (m, 1H) 3.60-3.71 (m, 180H) 3.48-3.60 (m, 4H) 3.35-3.44 (m, 5H) 2.23-2.37 (m, 4H) 1.62-1.86 (m, 6H) 1.21-1.37 (m, 40H) 0.83-0.93 (m, 6H).
  • EXAMPLE 44A was prepared using the same procedure described for EXAMPLE 35D, substituting hexadecanoic acid for tetradecanoic acid.
  • EXAMPLE 44B was prepared using the same procedure described for EXAMPLE 35E, substituting EXAMPLE 44A for EXAMPLE 35D. MS (ESI) m/z 482.6 (M+H) + .
  • EXAMPLE 44C was prepared using the same procedure described for EXAMPLE 35F, substituting EXAMPLE 44B for EXAMPLE 35E.
  • MS (MALDI) m/z 2689.0; 1 H NMR (300 MHz, CDCl 3 ) ⁇ ppm 5.09-5.19 (m, 1H) 4.17-4.26 (m, 3H) 4.01-4.11 (m, 1H) 3.73-3.91 (m, 1H) 3.61-3.70 (m, 180H) 3.48-3.60 (m, 4H) 3.35-3.44 (m, 5H) 2.23-2.36 (m, 4H) 1.54- 1.84 (m, 6H) 1.21-1.36 (m, 48H) 0.82-0.93 (m, 6H).
  • EXAMPLE 37B (100 mg) and mPEG-COOH (278 mg, PSA-288, Creative PEGWorks) were combined in dichloromethane (2 rnL). N 1 -((ethylimino)methylene)-N 3 ,N 3 - dimethylpropane- 1,3 -diamine hydrochloride (346 mg) was added followed by 4- (dimethylamino)pyridine (2 mg). The mixture was stirred overnight at room temperature then loaded directly onto a 4 g silica gel column (Analogix) and purified (Analogix 280, dichloromethane:methanol 0-20%).
  • N 1 -((ethylimino)methylene)-N 3 ,N 3 - dimethylpropane- 1,3 -diamine hydrochloride (346 mg) was added followed by 4- (dimethylamino)pyridine (2 mg). The mixture was stirred overnight at room temperature then loaded directly onto a 4 g silica gel
  • EXAMPLE 47A l-(2,2-dimethoxy-3-(tetradecyloxy)propoxy)tetradecane To a solution of 2,2-dimethoxypropane-1,3-diol (1 g) in toluene (30 mL) at 0°C was added NaH (1.484 g). The mixture was stirred at room temperature for 1 hour. The mixture was cooled to 0°C, and 1-bromotetradecane (4.99 mL) was added. The mixture was heated at reflux for 2 hours. The mixture was cooled to 0°C, and ethanol was added until it became clear. The mixture was concentrated. The concentrate was taken up in dichloromethane and dried onto silica gel.
  • the silica gel was loaded into an Analogix DASI module, and the product was isolated by flash chromatography (Analogix, SF65 ⁇ 200g, 2% ethyl acetate/hexanes for six column volumes, then 4% ethyl acetate/hexanes until the product eluted.
  • MS (ESI) m/z 500.4 (M+18) + .
  • 1,3-bis(tetradecyloxy)propan-2-yl 4-nitrophenyl carbonate To a solution of 1,3-bis(tetradecyloxy)propan-2-ol (0.3 g) in dichloromethane (3 mL) at 0°C were added triethlyamine (0.129 mL) and 4-nitrophenyl carbonochloridate (0.137 g). The mixture was stirred at room temperature overnight and concentrated. The concentrate was purified by flash chromatography (1 :10 ethyl acetate/hexanes).
  • This EXAMPLE was prepared as described in EXAMPLE 47, substituting hexadecyl methanesulfonate for 1-bromotetradecane in EXAMPLE 47A.
  • 1 H NMR 300 MHz, CDCl 3 ) ⁇ ppm 3.54-3.66 (m, 180H), 3.32-3.49 (m, 9H), 3.38 (s, 3H), 1.51-1.59 (m, 4H), 1.21-1.36 (m, 48H), 0.86-0.90 (m, 6H); MS (MALDI) m/z 2614.
  • This EXAMPLE was prepared as described in EXAMPLE 47 substituting octadecyl methanesulfonate for 1-bromotetradecane in EXAMPLE 47A.
  • 1 H NMR 300 MHz, CDCl 3 ) ⁇ ppm 3.52-3.66 (m, 180H), 3.32-3.49 (m, 9H), 3.38 (s, 3H), 1.51-1.59 (m, 4H), 1.21-1.36 (m, 52H), 0.86-0.90 (m, 6H); MS (MALDI) m/z 2557.
  • EXAMPLE 53A In a 100 mL round-bottomed flask was added N-Boc-serinol (1,1-dimethylethyl (2- hydroxy-1-(hydroxymethyl)ethyl)carbamate) (2.0 g) and sodium hydride (1.255 g) in N 5 N- dimethylformamide (50 mL). The mixture was cooled using an ice/water bath, and 1- bromohexadecane (7.98 g) was added to it. The mixture was heated at 70°C overnight, then cooled to room temperature. The mixture was cooled to 0°C and quenched with a few drops of cold water. The mixture was diluted with saturated ammonium chloride (50 mL).

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Abstract

L'invention concerne des lipides cationiques, des systèmes d'administration de médicaments à base de lipides cationiques, des façons de les fabriquer et des procédés de traitement de maladies avec ceux-ci.
PCT/US2009/040817 2008-04-16 2009-04-16 Lipides cationiques et utilisations de ceux-ci WO2009129395A1 (fr)

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Cited By (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010030739A1 (fr) * 2008-09-10 2010-03-18 Abbott Laboratories Conjugués lipidiques du polyéthylène glycol et leurs utilisations
WO2010030730A1 (fr) * 2008-09-10 2010-03-18 Abbott Laboratories Conjugués polyéthylène glycol-lipide et leurs utilisations
WO2011000835A3 (fr) * 2009-06-30 2011-03-03 Justus-Liebig-Universität Giessen Liposomes destinés à une application pulmonaire
WO2014007398A1 (fr) 2012-07-06 2014-01-09 協和発酵キリン株式会社 Lipide cationique
US8652512B2 (en) 2002-04-04 2014-02-18 United Therapeutics Corporation Nebulized liposomes for the pulmonary application of drug compounds
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US9352485B2 (en) 2012-03-23 2016-05-31 Empire Technology Development Llc Dioxaborinanes and uses thereof
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US10857105B2 (en) 2017-03-15 2020-12-08 MordernaTX, Inc. Compounds and compositions for intracellular delivery of therapeutic agents
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US11583504B2 (en) 2016-11-08 2023-02-21 Modernatx, Inc. Stabilized formulations of lipid nanoparticles
US11969506B2 (en) 2017-03-15 2024-04-30 Modernatx, Inc. Lipid nanoparticle formulation
US12077501B2 (en) 2017-06-14 2024-09-03 Modernatx, Inc. Compounds and compositions for intracellular delivery of agents

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4166132A (en) * 1977-08-18 1979-08-28 Pfizer Inc. Antiviral amine derivatives of glycerol and propanediols
US4255426A (en) * 1979-07-09 1981-03-10 Pfizer Inc. 1-(2-Hydroxy-3-n-alkoxypropyl)-4-substituted-piperazines and piperidines
US5723497A (en) * 1993-03-17 1998-03-03 Kao Corporation Amine derivative and dermatologic preparation containing the same
US6177469B1 (en) * 1995-10-14 2001-01-23 Harald Zilch Lipid alcohols as new immunosuppressive and antiviral drugs
WO2002003959A1 (fr) * 2000-07-06 2002-01-17 The General Hospital Corporation Preparations et ciblage d'apport de medicament
US6670332B1 (en) * 1995-11-30 2003-12-30 Vical Incorporated Complex cationic lipids having quarternary nitrogens therein
WO2005121348A1 (fr) * 2004-06-07 2005-12-22 Protiva Biotherapeutics, Inc. Arn interferant encapsule dans des lipides
US20060039942A1 (en) * 2004-08-20 2006-02-23 Greten Zachariah C Enhanced activity alcohol-based antimicrobial compositions

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4166132A (en) * 1977-08-18 1979-08-28 Pfizer Inc. Antiviral amine derivatives of glycerol and propanediols
US4255426A (en) * 1979-07-09 1981-03-10 Pfizer Inc. 1-(2-Hydroxy-3-n-alkoxypropyl)-4-substituted-piperazines and piperidines
US5723497A (en) * 1993-03-17 1998-03-03 Kao Corporation Amine derivative and dermatologic preparation containing the same
US6177469B1 (en) * 1995-10-14 2001-01-23 Harald Zilch Lipid alcohols as new immunosuppressive and antiviral drugs
US6670332B1 (en) * 1995-11-30 2003-12-30 Vical Incorporated Complex cationic lipids having quarternary nitrogens therein
WO2002003959A1 (fr) * 2000-07-06 2002-01-17 The General Hospital Corporation Preparations et ciblage d'apport de medicament
WO2005121348A1 (fr) * 2004-06-07 2005-12-22 Protiva Biotherapeutics, Inc. Arn interferant encapsule dans des lipides
US20060039942A1 (en) * 2004-08-20 2006-02-23 Greten Zachariah C Enhanced activity alcohol-based antimicrobial compositions

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
DATABASE BEILSTEIN [online] BEILSTEIN INSTITUTE FOR ORGANIC CHEMISTRY, FRANKFURT-MAIN, DE; XP002535770, Database accession no. BRN:10049676 *

Cited By (57)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8652512B2 (en) 2002-04-04 2014-02-18 United Therapeutics Corporation Nebulized liposomes for the pulmonary application of drug compounds
WO2010030739A1 (fr) * 2008-09-10 2010-03-18 Abbott Laboratories Conjugués lipidiques du polyéthylène glycol et leurs utilisations
WO2010030730A1 (fr) * 2008-09-10 2010-03-18 Abbott Laboratories Conjugués polyéthylène glycol-lipide et leurs utilisations
WO2011000835A3 (fr) * 2009-06-30 2011-03-03 Justus-Liebig-Universität Giessen Liposomes destinés à une application pulmonaire
US10258570B2 (en) 2009-06-30 2019-04-16 Lung Biotechnology Inc. Liposomes for pulmonary administration
US11246933B1 (en) 2011-12-07 2022-02-15 Alnylam Pharmaceuticals, Inc. Biodegradable lipids for the delivery of active agents
US11400158B2 (en) 2011-12-07 2022-08-02 Alnylam Pharmaceuticals, Inc. Biodegradable lipids for the delivery of active agents
US11382979B2 (en) 2011-12-07 2022-07-12 Alnylam Pharmaceuticals, Inc. Biodegradable lipids for the delivery of active agents
US11679158B2 (en) 2011-12-07 2023-06-20 Alnylam Pharmaceuticals, Inc. Biodegradable lipids for the delivery of active agents
US11633479B2 (en) 2011-12-07 2023-04-25 Alnylam Pharmaceuticals, Inc. Biodegradable lipids for the delivery of active agents
US11633480B2 (en) 2011-12-07 2023-04-25 Alnylam Pharmaceuticals, Inc. Biodegradable lipids for the delivery of active agents
US11612657B2 (en) 2011-12-07 2023-03-28 Alnylam Pharmaceuticals, Inc. Biodegradable lipids for the delivery of active agents
US11590229B2 (en) 2011-12-07 2023-02-28 Alnylam Pharmaceuticals, Inc. Biodegradable lipids for the delivery of active agents
US9352485B2 (en) 2012-03-23 2016-05-31 Empire Technology Development Llc Dioxaborinanes and uses thereof
WO2014007398A1 (fr) 2012-07-06 2014-01-09 協和発酵キリン株式会社 Lipide cationique
US9120938B2 (en) 2012-07-31 2015-09-01 Empire Technology Development Llc Polymerizable organoboron alkyd resin anti fouling coatings
US10323076B2 (en) 2013-10-03 2019-06-18 Modernatx, Inc. Polynucleotides encoding low density lipoprotein receptor
WO2015095340A1 (fr) * 2013-12-19 2015-06-25 Novartis Ag Lipides et compositions lipidiques pour le largage d'agents actifs
US11420933B2 (en) 2013-12-19 2022-08-23 Novartis Ag Lipids and lipid compositions for the delivery of active agents
EP3623361A1 (fr) * 2013-12-19 2020-03-18 Novartis AG Lipides et compositions de lipides pour l'administration d'agents actifs
US10059655B2 (en) 2013-12-19 2018-08-28 Novartis Ag Lipids and lipid compositions for the delivery of active agents
EP3872066A1 (fr) * 2013-12-19 2021-09-01 Novartis AG Lipides et compositions lipidiques pour l'administration d'agents actifs
US11013696B2 (en) 2013-12-19 2021-05-25 Novartis Ag Lipids and lipid compositions for the delivery of active agents
US10426737B2 (en) 2013-12-19 2019-10-01 Novartis Ag Lipids and lipid compositions for the delivery of active agents
US10906867B2 (en) 2013-12-19 2021-02-02 Novartis Ag Lipids and lipid compositions for the delivery of active agents
US10342761B2 (en) 2014-07-16 2019-07-09 Novartis Ag Method of encapsulating a nucleic acid in a lipid nanoparticle host
US9868691B2 (en) 2015-09-17 2018-01-16 Modernatx, Inc. Compounds and compositions for intracellular delivery of therapeutic agents
JP7326395B2 (ja) 2015-09-17 2023-08-15 モデルナティエックス インコーポレイテッド 治療剤の細胞内送達のための化合物および組成物
US9868693B2 (en) 2015-09-17 2018-01-16 Modernatx, Inc. Compounds and compositions for intracellular delivery of therapeutic agents
US9868692B2 (en) 2015-09-17 2018-01-16 Modernatx, Inc. Compounds and compositions for intracellular delivery of therapeutic agents
EP3736261A1 (fr) * 2015-09-17 2020-11-11 ModernaTX, Inc. Composés et compositions pour l'administration intracellulaire d'agents thérapeutiques
WO2017049245A3 (fr) * 2015-09-17 2017-05-18 Modernatx, Inc. Composés et compositions pour l'administration intracellulaire d'agents thérapeutiques
EP4286012A3 (fr) * 2015-09-17 2024-05-29 ModernaTX, Inc. Composés et compositions pour l'administration intracellulaire d'agents thérapeutiques
US10442756B2 (en) 2015-09-17 2019-10-15 Modernatx, Inc. Compounds and compositions for intracellular delivery of therapeutic agents
US10392341B2 (en) 2015-09-17 2019-08-27 Modernatx, Inc. Compounds and compositions for intracellular delivery of therapeutic agents
US9867888B2 (en) 2015-09-17 2018-01-16 Modernatx, Inc. Compounds and compositions for intracellular delivery of therapeutic agents
US10266485B2 (en) 2015-09-17 2019-04-23 Modernatx, Inc. Compounds and compositions for intracellular delivery of therapeutic agents
JP2018532721A (ja) * 2015-09-17 2018-11-08 モデルナティエックス インコーポレイテッドＭｏｄｅｒｎａＴＸ，Ｉｎｃ． 治療剤の細胞内送達のための化合物および組成物
US11220476B2 (en) 2015-09-17 2022-01-11 Modernatx, Inc. Compounds and compositions for intracellular delivery of therapeutic agents
JP2022008431A (ja) * 2015-09-17 2022-01-13 モデルナティエックス インコーポレイテッド 治療剤の細胞内送達のための化合物および組成物
JP6948313B6 (ja) 2015-09-17 2022-01-14 モデルナティエックス インコーポレイテッド 治療剤の細胞内送達のための化合物および組成物
AU2016377681B2 (en) * 2015-12-22 2021-05-13 Modernatx, Inc. Compounds and compositions for intracellular delivery of agents
US10195156B2 (en) 2015-12-22 2019-02-05 Modernatx, Inc. Compounds and compositions for intracellular delivery of agents
WO2017112865A1 (fr) * 2015-12-22 2017-06-29 Modernatx, Inc. Composés et compositions pour l'administration intracellulaire d'agents thérapeutiques et/ou prophylactiques
US10799463B2 (en) 2015-12-22 2020-10-13 Modernatx, Inc. Compounds and compositions for intracellular delivery of agents
US11583504B2 (en) 2016-11-08 2023-02-21 Modernatx, Inc. Stabilized formulations of lipid nanoparticles
US11203569B2 (en) 2017-03-15 2021-12-21 Modernatx, Inc. Crystal forms of amino lipids
US11969506B2 (en) 2017-03-15 2024-04-30 Modernatx, Inc. Lipid nanoparticle formulation
US10857105B2 (en) 2017-03-15 2020-12-08 MordernaTX, Inc. Compounds and compositions for intracellular delivery of therapeutic agents
WO2018177383A1 (fr) * 2017-03-29 2018-10-04 中国医学科学院基础医学研究所 Application d'un composé ou d'un extrait de médecine chinoise traditionnelle dans la préparation d'un agent d'administration d'acide nucléique et produits apparentés correspondants
US12077501B2 (en) 2017-06-14 2024-09-03 Modernatx, Inc. Compounds and compositions for intracellular delivery of agents
WO2019027055A1 (fr) 2017-08-04 2019-02-07 協和発酵キリン株式会社 Nanoparticules lipidiques contenant un acide nucléique
CN111918858B (zh) * 2018-03-29 2023-03-21 中国医学科学院基础医学研究所 化合物或中药提取物在制备核酸递送试剂中的应用及其相关产品
WO2019184991A1 (fr) * 2018-03-29 2019-10-03 中国医学科学院基础医学研究所 Utilisation d'un composé ou d'un extrait de médecine chinoise traditionnelle dans la préparation d'un agent d'administration d'acide nucléique, et produits associés
CN111918858A (zh) * 2018-03-29 2020-11-10 中国医学科学院基础医学研究所 化合物或中药提取物在制备核酸递送试剂中的应用及其相关产品
US11597698B2 (en) 2019-09-19 2023-03-07 Modernatx, Inc. Branched tail lipid compounds and compositions for intracellular delivery of therapeutic agents
US11066355B2 (en) 2019-09-19 2021-07-20 Modernatx, Inc. Branched tail lipid compounds and compositions for intracellular delivery of therapeutic agents

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