WO2009129387A2 - Lipides cationiques et utilisations associees - Google Patents

Lipides cationiques et utilisations associees Download PDF

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Publication number
WO2009129387A2
WO2009129387A2 PCT/US2009/040803 US2009040803W WO2009129387A2 WO 2009129387 A2 WO2009129387 A2 WO 2009129387A2 US 2009040803 W US2009040803 W US 2009040803W WO 2009129387 A2 WO2009129387 A2 WO 2009129387A2
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bis
octadeca
dienyloxy
butyl
weight
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PCT/US2009/040803
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English (en)
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WO2009129387A3 (fr
Inventor
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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Publication of WO2009129387A2 publication Critical patent/WO2009129387A2/fr
Publication of WO2009129387A3 publication Critical patent/WO2009129387A3/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/543—Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6905—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion
    • A61K47/6911—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion the form being a liposome
    • 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
    • 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
    • A61K9/1277—Processes for preparing; Proliposomes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C235/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
    • C07C235/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/04—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C235/08—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton being acyclic and saturated having the nitrogen atom of at least one of the carboxamide groups bound to an acyclic carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C271/00—Derivatives of carbamic acids, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
    • C07C271/06—Esters of carbamic acids
    • C07C271/08—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms
    • C07C271/10—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C271/16—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by singly-bound oxygen atoms
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D203/00—Heterocyclic compounds containing three-membered rings with one nitrogen atom as the only ring hetero atom
    • C07D203/04—Heterocyclic compounds containing three-membered rings with one nitrogen atom as the only ring hetero atom not condensed with other rings
    • C07D203/06—Heterocyclic compounds containing three-membered rings with one nitrogen atom as the only ring hetero atom not condensed with other rings having no double bonds between ring members or between ring members and non-ring members
    • C07D203/08—Heterocyclic compounds containing three-membered rings with one nitrogen atom as the only ring hetero atom not condensed with other rings having no double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to the ring nitrogen atom
    • C07D203/10—Radicals substituted by singly bound oxygen atoms
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • 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
    • C07D207/06—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 radicals, containing only hydrogen and carbon atoms, attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings 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
    • C07D211/36—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings 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
    • C07D211/40—Oxygen atoms
    • C07D211/44—Oxygen atoms attached in position 4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings 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
    • C07D211/36—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings 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
    • C07D211/56—Nitrogen atoms
    • C07D211/58—Nitrogen atoms attached in position 4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/56—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
    • C07D233/61—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 nitrogen atoms not forming part of a nitro radical, attached to ring nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/08—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms
    • C07D295/084—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms with the ring nitrogen atoms and the oxygen or sulfur atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings
    • C07D295/088—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms with the ring nitrogen atoms and the oxygen or sulfur atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings to an acyclic saturated chain
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • 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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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/88—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation using microencapsulation, e.g. using amphiphile liposome vesicle

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)
  • Y is CH 2 , NH or O
  • R and R are independently H, cycloalkyl, cycloalkenyl or R ; or
  • R and R together with the nitrogen to which they are attached, are heterocycloalkyl or heteroaryl;
  • R and R is H, and the other is C 14 -C 2 o-alkenyl, or C 14 -C 2 o-alkyl; or
  • R and R are independently selected C 14 -C 2 o-alkenyl, or C 14 -C 2 o-alkyl; or
  • R and R together are CR R , wherein R is H and R is C 14 -C 2 o-alkenyl, C 14 - C 2 o-alkyl, or CH 2 ⁇ -C 14 -C 2 o-alkenyl; or R and R are independently selected 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 , NR 6 S(O) 2 R 6 , NHC(O)OR 6 , NR 6 C(O)OR 6 , NHC(O)NH 2 ,
  • 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 ⁇ 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 ⁇ 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 16E , 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
  • R is alkyl, alkenyl, alkynyl, or R ;
  • R is phenyl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl;
  • 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.
  • 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, II, III, or IV.
  • CaBLES Cationic-Based Lipid Encapsulation Systems
  • cationic lipids of the present invention i.e., cationic lipids of Formula I, II, III or IV
  • cationic lipids of Formula I, II, III or IV 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 antisense oligonucleotide
  • DNA a DNA
  • a plasmid a ribosomal RNA
  • miRNA micro RNA
  • tRNA transfer RNA
  • siRNA small inhibitory RNA
  • snRNA small nuclear RNA
  • antigens fragments thereof, proteins, peptides, vaccines and small-molecules or mixtures thereof
  • a further embodiment pertains to pharmaceutical compositions comprising a Lipid- Based Particle and a pharmaceutically acceptable carrier.
  • 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 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).
  • Figures 1-48 In vivo delivery and vitro transfection activity of selected cationic lipids that were formulated as disclosed herein.
  • 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.
  • Ci-C ⁇ -alkylene means divalent, saturated, straight or branched chain hydrocarbon moieties bonds, such as Ci-alkylene, C2-alkylene, C3-alkylene, C4-alkylene, Cs-alkylene and C ⁇ -alkylene.
  • alkyl as used herein, means monovalent, straight or branched chain hydrocarbon moieties such as Ci-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.
  • d-Cg-alkyl as used herein, means Ci-alkyl, C 2 -alkyl, C 3 -alkyl, C 4 -alkyl, Cs-alkyl, C 6 -alkyl, C 7 -alkyl and C 8 -alkyl.
  • Ci 4 -C 2 o-alkenyl means Ci 4 -alkenyl,” C 15 - alkenyl,” Ci 6 - alkenyl,” C n - alkenyl,” Ci 8 - alkenyl,” C 19 - alkenyl” and C 20 - alkenyl.”
  • C 14 -C 2 o-alkyl means C 14 -alkyl,” C 15 -alkyl,” C 16 -alkyl,” Civ-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 means l,2-distearoyl-5/?-glycero-3-phosphocholine.
  • Choi means cholesterol.
  • PEG-Chol means poly(oxy-l,2-ethanediyl)-2000- ⁇ -(3 ⁇ )- cholest-S-en-S-yl-omega-hydroxy.
  • Pal-PEG-Cera N-palmitoyl-sphingosine-1- [succinyl(methoxypolyethylene glycol)-2000] .
  • PEG-DMPE N-(carbonyl- methoxypolyethyleneglycol-2000)-l,2-dimyristoyl-sn-glycero-3-phosphoethanolamine.
  • PEG-DPPE N-(carbonyl- methoxypolyethyleneglycol-2000)-l,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine.
  • PEG-DSPE N-(carbonyl- methoxypolyethyleneglycol-2000)-l,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.
  • SPC soybean phosphatidylcholine
  • MALDI matrix assisted laser desorption ionization
  • nanoparticle 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.
  • 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)
  • Y is CH 2 , NH or O
  • Y is a bond or C(O);
  • Y is a bond or C 1 -C 6 alky lene; 1 2 5
  • R and R are independently H, cycloalkyl, cycloalkenyl or R ; or
  • R and R together with the nitrogen to which they are attached, are heterocycloalkyl or heteroaryl;
  • R and R is H, and the other is C 14 -C 2 o-alkenyl, or C 14 -C 2 o-alkyl; or
  • R and R are independently selected C 14 -C 2 o-alkenyl, or C 14 -C 2 o-alkyl; or
  • C 2 o-alkyl or CH 2 O-C 14 -C 2 o-alkenyl; or R and R are independently selected C 14 -C 2 o-alkenyl, C 14 -C 2 o-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 , NR 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)NHOH, C(O)NHOR 6 , C(O)NHSO 2 R 6 , C(O)NR 6 SO 2 R 6 , SO 2 NH 2 , SO 2 NHR 6 , SO 2 N(R 6 ) 2 , C(O)H, C(O)OH, C(N)NH 2 , C(N)NHR 6 , C(N)N(R 6 ) 2 , CNOH, CNO
  • 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 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 ⁇ 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 ⁇ 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 ) 2 , C(O)
  • 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 16E , 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
  • R is alkyl, alkenyl, alkynyl, or R ;
  • R is phenyl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl;
  • 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
  • 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)
  • Y is NH or O
  • Y is a bond or C 1 -C 6 alky lene
  • R and R are independently H or R ;
  • R and R together with the nitrogen to which they are attached, are heterocycloalkyl;
  • R and R are independently selected C 14 -C 2 o-alkenyl, or C 14 -C 2 o-alkyl; or 3 4 20 21 20 21
  • R and R together are CR R , wherein R and R are independently selected C 14 -C 2 o-alkenyl;
  • R is alkyl, which is unsubstituted or substituted with one or more R , OR , or N(R 6 ) 2 ;
  • R 6 is R 7 , R 8 , R 9 , or R 10 ;
  • R is phenyl
  • R is heteroaryl
  • R is heterocycloalkyl
  • R 10 is alkyl
  • each foregoing cyclic moiety is independently unsubstituted or substituted with one or more R , OR , N(R ) 2 , or F, Cl, Br or I;
  • R is R , R , or R ;
  • R is phenyl
  • R is heteroaryl
  • R is alkyl
  • R and R are independently unsubstituted or substituted with OR ;
  • R 18 is alkyl
  • One embodiment of this invention therefore pertains to a cationic lipid or mixtures thereof, having Formula (I)
  • Y 3 and Y 4 are each a bond. Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each C(O). Another embodiment of Formula (I) pertains to compounds wherein Y 3 is a bond and Y 4 is C(O). Another embodiment of Formula (I) pertains to compounds wherein Y 4 is a bond and Y 3 is C(O).
  • Formula (I) pertains to compounds wherein R 1 and R 2 are each R 5 . Another embodiment of Formula (I) pertains to compounds wherein R 1 is H and R 2 is R 5 . Another embodiment of Formula (I) pertains to compounds wherein R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl. Another embodiment of Formula (I) pertains to compounds wherein R 1 and R 2 together with the nitrogen to which they are attached, are heteroaryl. Another embodiment of Formula (I) pertains to compounds wherein R 1 and R 2 together with the nitrogen to which they are attached, are pyrrolidinyl, piperidinyl, morpholinyl, or piperazinyl. Another embodiment of Formula (I) pertains to compounds wherein R 1 and R 2 together with the nitrogen to which they are attached, are pyrrolidinyl.
  • One embodiment of Formula (I) pertains to compounds wherein R 3 and R 4 are C 14 -
  • Formula (I) pertains to compounds wherein R 5 is alkyl. Another embodiment of Formula (I) pertains to compounds wherein R 5 is alkyl which is unsubstituted. Another embodiment of Formula (I) pertains to compounds wherein R 5 is alkyl which is substituted. Another embodiment of Formula (I) pertains to compounds wherein R 5 is alkyl which is substituted with R 6 , OR 6 , or N(R 6 ) 2 .
  • Formula (I) pertains to compounds wherein R 6 is R 7 ; and R 7 is phenyl which is unfused. Another embodiment of Formula (I) pertains to compounds wherein R 6 is R 8 ; and R 8 is heteroaryl, which is unfused. Another embodiment of Formula (I) pertains to compounds wherein R 6 is R 9 ; and R 9 is heterocycloalkyl, which is unfused. Another embodiment of Formula (I) pertains to compounds wherein R 6 is R 10 ; and R 10 is alkyl, which is unsubstituted.
  • One embodiment of Formula (I) pertains to compounds wherein all foregoing cyclic moieties are unsubstituted. Another embodiment of Formula (I) pertains to compounds wherein one or more cyclic moieties are substituted. Another embodiment of Formula (I) pertains to compounds wherein one or more cyclic moieties are substituted with one or more R 11 , OR 11 , or N(R 1 %, or F.
  • One embodiment pertains to compounds of Formula (I) wherein Y 3 and Y 4 are each a bond; R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl; and R 3 and R 4 are Ci 4 -C 2 o-alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 are each independently R 5 ; R 5 is alkyl which is unsubstituted; and R 3 and R 4 are each Ci 4 -C 2 o-alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl; R 3 and R 4 are C14-C20- alkenyl; wherein the heterocycloalkyl is substituted with R 11 ; R 11 is R 12 ; and R 12 is phenyl which is unfused.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl; R 3 and R 4 are Ci 4 -C 2 o-alkenyl; wherein the heterocycloalkyl is substituted with R 11 ; R 11 is R 15 ; and R 15 is alkyl which is unsubstituted.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 are each independently R 5 ; one R 5 is alkyl which is unsubstituted, and the other R 5 is alkyl which is substituted with one OR 6 ; R 6 is R 10 ; R 10 is alkyl; and R 3 and R 4 are each Ci 4 -C 2 o-alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl; R 3 and R 4 are Ci 4 -C 2 o-alkenyl; wherein the heterocycloalkyl is substituted with R 11 ; R 11 is R ; R is phenyl which is unfused; wherein R is substituted with one OR ; and R is alkyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 are each independently R 5 ; one R 5 is alkyl which is unsubstituted, and the other R 5 is alkyl which is substituted with one N(R 6 ) 2 ; R 6 is R 10 ; R 10 is alkyl; and R 3 and R 4 are each Ci 4 -C 2 o-alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 are each independently R 5 ; one R 5 is alkyl which is unsubstituted, and the other R 5 is alkyl which is substituted with one R 6 ; R 6 is R 8 ; R 8 is heteroaryl which is unfused; and R 3 and R 4 are each Ci 4 -C 2 o-alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 are each independently R 5 ; one R 5 is alkyl which is unsubstituted, and the other R 5 is alkyl which is substituted with one R 6 ; R 6 is R 7 ; R 7 is phenyl which is unfused; and R 3 and R 4 are each C 14 - C 2 o-alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 are each independently R 5 ; one R 5 is alkyl which is unsubstituted, and the other R 5 is alkyl which is substituted with one R 6 ; R 6 is R 7 ; R 7 is phenyl which is unfused; and R 3 and R 4 are each Ci4-C2o-alkenyl; wherein R 7 is substituted with one F.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl; R 3 and R 4 are Ci 4 -C 2 o-alkenyl; wherein the heterocycloalkyl is substituted with R 11 ; R 11 is R 12 ; and R 12 is phenyl which is unfused; wherein R 12 is substituted with one F.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl; R 3 and R 4 are Ci4-C2o-alkenyl; wherein the heterocycloalkyl is substituted with N(R ⁇ )2; R 11 is R 15 ; and R 15 is alkyl which is unsubstituted.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 are each independently R 5 ; each R 5 is alkyl which is substituted with one OR 6 ; R 6 is R 10 ; R 10 is alkyl; and R 3 and R 4 are each C 14 -C 20 -alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl; R 3 and R 4 are Ci 4 -C 2 o-alkenyl; wherein the heterocycloalkyl is substituted with OR 11 ; R 11 is R 15 ; and R 15 is alkyl which is unsubstituted.
  • R 1 and R 2 together with the nitrogen to which they are attached are heterocycloalkyl
  • R 3 and R 4 are Ci 4 -C 2 o-alkenyl
  • the heterocycloalkyl is substituted with OR 11 ;
  • R 11 is R 15 ; and R 15 is alkyl which is unsubstituted.
  • Formula (I) pertains to compounds wherein Y 3 and Y 4 are each C(O); R 1 and R 2 are each independently R 5 ; R 5 is alkyl which is unsubstituted; and R 3 and R 4 are each Ci 4 -C 2 o-alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 are each independently R 5 ; R 5 is alkyl which is unsubstituted; and R 3 and R 4 together are CR 20 R 21 , wherein R 20 and R 21 are each Ci 4 -C 2 o-alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each C(O); R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl; and R 3 and R 4 are C14-C20- alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl; and R 3 and R 4 are Ci 4 -C 2 o-alkyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 is H; R 2 is R 5 ; R 5 is alkyl which is substituted with R 6 ; R 6 is R 8 ; R 8 is heteroaryl which is unfused; and R 3 and R 4 are Ci 4 -C 2 o-alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 is H; R 2 is R 5 ; R 5 is alkyl which is substituted with R 6 ; R 6 is R 8 ; R 8 is heteroaryl which is unfused; and R 3 and R 4 are Ci 4 -C 2 o-alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and
  • Y 4 are each a bond; R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl; R 3 and R 4 are Ci 4 -C 2 o-alkenyl; wherein the heterocycloalkyl is substituted with R 11 ; R 11 is R 13 ; and R 13 is heteroaryl which is unfused.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 is H; R 2 is R 5 ; R 5 is alkyl which is substituted with R 6 ; R 6 is R 9 ; R 9 is heterocycloalkyl which is unfused; and R 3 and R 4 are Ci 4 -C 2 o-alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 is H; R 2 is R 5 ; R 5 is alkyl which is substituted with N(R 6 ) 2 ; R 6 is R 10 ; R 10 is alkyl; and R 3 and R 4 are Ci 4 -C 2 o-alkenyl.
  • Another embodiment of Formula (I) pertains to compounds wherein Y 3 and Y 4 are each a bond; R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl; R 3 and R 4 are Ci 4 -C 2 o-alkenyl; wherein the heterocycloalkyl is substituted with two R 11 ; each R 11 is R 15 ; and each R 15 is alkyl which is unsubstituted.
  • One embodiment of this invention therefore pertains to a cationic lipid or mixtures thereof, having Formula (II)
  • Y 3 and Y 4 are each a bond.
  • Another embodiment of Formula (II) pertains to compounds wherein Y 3 and Y 4 are each C(O).
  • Another embodiment of Formula (II) pertains to compounds wherein Y 3 is a bond and Y 4 is C(O).
  • Another embodiment of Formula (II) pertains to compounds wherein Y 4 is a bond and Y 3 is C(O).
  • One embodiment of Formula (II) pertains to compounds wherein R 1 and R 2 are each R 5 .
  • Another embodiment of Formula (II) pertains to compounds wherein R 1 is H and R 2 is R 5 .
  • Another embodiment of Formula (II) pertains to compounds wherein R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl.
  • Another embodiment of Formula (II) pertains to compounds wherein R 1 and R 2 together with the nitrogen to which they are attached, are heteroaryl. Another embodiment of Formula (II) pertains to compounds wherein R 1 and R 2 together with the nitrogen to which they are attached, are pyrrolidinyl, azetidinyl, or piperazinyl. Another embodiment of Formula (II) pertains to compounds wherein R 1 and R 2 together with the nitrogen to which they are attached, are pyrrolidinyl.
  • Formula (II) pertains to compounds wherein R 3 and R 4 are C 14 - C 2 o-alkenyl. Another embodiment of Formula (II) pertains to compounds wherein R 3 and R 4 are Ci 4 -C 2 o-alkyl. Another embodiment of Formula (II) pertains to compounds wherein R 3 and R 4 together are CR 20 R 21 , wherein R 20 and R 21 are each Ci 4 -C 2 o-alkenyl. Another embodiment of Formula (II) pertains to compounds wherein R 3 is H; and R 4 is C14-C20- alkenyl.
  • Formula (II) pertains to compounds wherein R 5 is alkyl. Another embodiment of Formula (II) pertains to compounds wherein R 5 is alkyl which is unsubstituted. Another embodiment of Formula (II) pertains to compounds wherein R 5 is alkyl which is substituted. Another embodiment of Formula (II) pertains to compounds wherein R 5 is alkyl which is substituted with R , OR , or N(R ) 2 .
  • Formula (II) pertains to compounds wherein R 6 is R 7 ; and R 7 is phenyl which is unfused. Another embodiment of Formula (II) pertains to compounds wherein R 6 is R 8 ; and R 8 is heteroaryl, which is unfused. Another embodiment of Formula (II) pertains to compounds wherein R 6 is R 9 ; and R 9 is heterocycloalkyl, which is unfused. Another embodiment of Formula (II) pertains to compounds wherein R 6 is R 10 ; and R 10 is alkyl, which is unsubstituted.
  • One embodiment of Formula (II) pertains to compounds wherein all foregoing cyclic moieties are unsubstituted. Another embodiment of Formula (II) pertains to compounds wherein one or more cyclic moieties are substituted. Another embodiment of Formula (II) pertains to compounds wherein one or more cyclic moieties are substituted with one or more R 11 , OR 11 , or N(R 1 %, or F.
  • One embodiment pertains to compounds of Formula (II) wherein Y 3 and Y 4 are each a bond; Y 1 is C 1 -C 6 alkylene; R 1 and R 2 are each independently R 5 ; each R 5 is alkyl which is unsubstituted; and R 3 and R 4 are Ci 4 -C 2 o-alkenyl.
  • Another embodiment of Formula (II) pertains to compounds wherein Y 3 and Y 4 are each a bond; Y 1 is C 1 -C 6 alkylene; R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl; R 3 and R 4 are C 14 - C 2 o-alkenyl; wherein the heterocycloalkyl is substituted with one R 11 ; R 11 is R 15 ; and R 15 is alkyl which is unsubstituted.
  • Formula (II) pertains to compounds wherein Y 3 and Y 4 are each a bond; Y 1 is C 1 -C 6 alkylene; R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl; and R 3 and R 4 are C 14 -C 2 o-alkenyl.
  • One embodiment of this invention therefore pertains to a cationic lipid or mixtures thereof, having Formula (III)
  • Y 2 is NH.
  • Another embodiment of Formula (III) pertains to compounds wherein Y 2 is O.
  • Another embodiment of Formula (III) pertains to compounds wherein Y 2 is CH 2 .
  • Formula (III) pertains to compounds wherein Y 3 and Y 4 are each a bond. Another embodiment of Formula (III) pertains to compounds wherein Y 3 and Y 4 are each C(O). Another embodiment of Formula (III) pertains to compounds wherein Y 3 is a bond and Y 4 is C(O). Another embodiment of Formula (III) pertains to compounds wherein Y 4 is a bond and Y 3 is C(O). One embodiment of Formula (III) pertains to compounds wherein Y 6 is a bond. Another embodiment of Formula (III) pertains to compounds wherein Y 6 is Ci-C 6 alkylene.
  • Formula (III) pertains to compounds wherein R 1 and R 2 are each R 5 .
  • Another embodiment of Formula (III) pertains to compounds wherein R 1 is H and R 2 is
  • R 5 Another embodiment of Formula (III) pertains to compounds wherein R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl. Another embodiment of Formula (III) pertains to compounds wherein R 1 and R 2 together with the nitrogen to which they are attached, are heteroaryl.
  • Formula (III) pertains to compounds wherein R 3 and R 4 are C 14 - C 2 o-alkenyl. Another embodiment of Formula (III) pertains to compounds wherein R 3 and R 4 are Ci 4 -C 2 o-alkyl. Another embodiment of Formula (III) pertains to compounds wherein R and R 4 together are CR 20 R 21 , wherein R 20 and R 21 are each Ci 4 -C 2 o-alkenyl. Another embodiment of Formula (III) pertains to compounds wherein R 3 is H; and R 4 is C14-C20- alkenyl.
  • Formula (III) pertains to compounds wherein R 5 is alkyl. Another embodiment of Formula (III) pertains to compounds wherein R 5 is alkyl which is unsubstituted. Another embodiment of Formula (III) pertains to compounds wherein R 5 is alkyl which is substituted. Another embodiment of Formula (III) pertains to compounds wherein R 5 is alkyl which is substituted with R , OR , or N(R ) 2 .
  • Formula (III) pertains to compounds wherein R 6 is R 7 ; and R 7 is phenyl which is unfused. Another embodiment of Formula (III) pertains to compounds wherein R 6 is R 8 ; and R 8 is heteroaryl, which is unfused. Another embodiment of Formula (III) pertains to compounds wherein R 6 is R 9 ; and R 9 is heterocycloalkyl, which is unfused. Another embodiment of Formula (III) pertains to compounds wherein R 6 is R 10 ; and R 10 is alkyl, which is unsubstituted. One embodiment of Formula (III) pertains to compounds wherein all foregoing cyclic moieties are unsubstituted.
  • Another embodiment of Formula (III) pertains to compounds wherein one or more cyclic moieties are substituted.
  • Another embodiment of Formula (III) pertains to compounds wherein one or more cyclic moieties are substituted with one or more R 11 , OR 11 , or N(R 1 %, or F.
  • Another embodiment of this invention therefore pertains to a cationic lipid or mixtures thereof, having Formula (IV)
  • Y 2 is NH.
  • Another embodiment of Formula (IV) pertains to compounds wherein Y 2 is O.
  • Another embodiment of Formula (IV) pertains to compounds wherein Y 2 is CH 2 .
  • Formula (IV) pertains to compounds wherein Y 3 and Y 4 are each a bond. Another embodiment of Formula (IV) pertains to compounds wherein Y 3 and Y 4 are each C(O). Another embodiment of Formula (IV) pertains to compounds wherein Y is a bond and Y 4 is C(O). Another embodiment of Formula (IV) pertains to compounds wherein Y 4 is a bond and Y 3 is C(O).
  • Formula (IV) pertains to compounds wherein Y 5 is CH 2 . Another embodiment of Formula (IV) pertains to compounds wherein Y 5 is NH. Another embodiment of Formula (IV) pertains to compounds wherein Y 5 is O.
  • Formula (IV) pertains to compounds wherein Y 6 is a bond.
  • Another embodiment of Formula (IV) pertains to compounds wherein Y 6 is Ci-C 6 alkylene.
  • Formula (IV) pertains to compounds wherein R 1 and R 2 are each R 5 . Another embodiment of Formula (IV) pertains to compounds wherein R 1 is H and R 2 is R 5 . Another embodiment of Formula (IV) pertains to compounds wherein R 1 and R 2 together with the nitrogen to which they are attached, are heterocycloalkyl. Another embodiment of Formula (IV) pertains to compounds wherein R 1 and R 2 together with the nitrogen to which they are attached, are heteroaryl. . Another embodiment of Formula (IV) pertains to compounds wherein R 1 and R 2 together with the nitrogen to which they are attached, is pyrrolidinyl.
  • Formula (IV) pertains to compounds wherein R 3 and R 4 are C 14 - C 2 o-alkenyl. Another embodiment of Formula (IV) pertains to compounds wherein R 3 and R 4 are Ci 4 -C 2 o-alkyl. Another embodiment of Formula (IV) pertains to compounds wherein R 3 and R 4 together are CR 20 R 21 , wherein R 20 and R 21 are each C 14 -C 20 -alkenyl. Another embodiment of Formula (IV) pertains to compounds wherein R 3 is H; and R 4 is C14-C20- alkenyl.
  • Formula (IV) pertains to compounds wherein R 5 is alkyl. Another embodiment of Formula (IV) pertains to compounds wherein R 5 is alkyl which is unsubstituted. Another embodiment of Formula (IV) pertains to compounds wherein R 5 is alkyl which is substituted. Another embodiment of Formula (IV) pertains to compounds wherein R 5 is alkyl which is substituted with R , OR , or N(R ) 2 .
  • Formula (IV) pertains to compounds wherein R 6 is R 7 ; and R 7 is phenyl which is unfused. Another embodiment of Formula (IV) pertains to compounds wherein R 6 is R 8 ; and R 8 is heteroaryl, which is unfused. Another embodiment of Formula (IV) pertains to compounds wherein R 6 is R 9 ; and R 9 is heterocycloalkyl, which is unfused. Another embodiment of Formula (IV) pertains to compounds wherein R 6 is R 10 ; and R 10 is alkyl, which is unsubstituted.
  • One embodiment of Formula (IV) pertains to compounds wherein all foregoing cyclic moieties are unsubstituted. Another embodiment of Formula (IV) pertains to compounds wherein one or more cyclic moieties are substituted. Another embodiment of Formula (IV) pertains to compounds wherein one or more cyclic moieties are substituted with one or more R 11 , OR 11 , or N(R 1 %, or F.
  • One embodiment pertains to compounds of Formula (IV) wherein Y 3 and Y 4 are each a bond; Y 2 is O; Y 5 is NH; Y 6 is Ci-C 6 alkylene; R 1 and R 2 together with the nitrogen to which they are attached are heterocycloalkyl; and R 3 and R 4 are Ci 4 -C 2 o-alkenylAnother embodiment of Formula (IV) pertains to compounds wherein Y 3 and Y 4 are each a bond; Y 2 is NH; Y 5 is CH 2 ; Y 6 is a bond; R 1 and R 2 are each independently R 5 ; each R 5 is alkyl which is unsubstituted; and R 3 and R 4 are C 14 -C 20 -alkenyl.
  • Still another embodiment pertains to compounds of this invention which include, but are not limited to l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ piperidine, 4- ⁇ 3,4- bis[(9Z, 12Z)-octadeca-9, 12-dienyloxy]butyl ⁇ morpholine, N,N-diethyl-3,4-bis[(9Z, 12Z)- octadeca-9,12-dienyloxy]butan-l -amine, N,N-dimethyl-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butan-l -amine, l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ -4- phenylpiperazine, 1 - ⁇ 3 ,4-bis[(9Z, 12Z
  • Still another embodiment pertains to compounds of this invention wherein one or more cationic lipids are chosen from l- ⁇ (3S)-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ pyrrolidine, l- ⁇ (3R)-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ pyrrolidine, l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ -N,N- dimethylpyrrolidin-3-amine, and l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ pyrrolidine .
  • Still another embodiment pertains to compounds of this invention wherein one or more cationic lipids are chosen from N,N-diethyl-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butan- 1 -amine, N,N-dimethyl-3 ,4-bis[(9Z, 12Z)-octadeca-9, 12-dienyloxy]butan- 1 - amine, N-(2-methoxyethyl)-N-methyl-3 ,4-bis[(9Z, 12Z)-octadeca-9, 12-dienyloxy]butan- 1 - amine, N- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ -N,N',N'-trimethylethane-l,2- diamine, 1 - ⁇ 3 ,4-bis[(9Z, 12Z)-octadeca
  • a further embodiment pertains to particles comprising one or more cationic lipid(s) having Formula I, II, III, or IV.
  • a further embodiment pertains to particles comprising one or more cationic lipid(s) having Formula I, II, III, or IV 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
  • a further embodiment pertains to nanoparticles comprising one or more cationic lipid(s) having Formula I, II, III, or IV.
  • a further embodiment pertains to nanoparticles comprising one or more cationic lipid(s) having Formula I, II, III, or IV 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 antisense oligonucleotide
  • DNA 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.
  • rRNA ribosomal RNA
  • miRNA micro RNA
  • tRNA transfer 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, II, III, or IV.
  • 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)
  • Y 2 is CH 2 , NH or O
  • Y 3 is a bond or C(O);
  • Y 4 is a bond or C(O);
  • Y 5 is CH 2 , NH or O
  • Y is a bond or C 1 -C 6 alky lene
  • R and R are independently H, cycloalkyl, cycloalkenyl or R ; or
  • R and R together with the nitrogen to which they are attached, are heterocycloalkyl or heteroaryl;
  • R and R is H, and the other is C 14 -C 2 o-alkenyl, or C 14 -C 2 o-alkyl; or
  • R and R are independently selected C 14 -C 2 o-alkenyl, or C 14 -C 2 o-alkyl; or
  • R and R together are CR R , wherein R is H and R is C 14 -C 2 o-alkenyl, C 14 - C 2 o-alkyl, or CH 2 O-C 14 -C 2 o-alkenyl; or R and R are independently selected
  • 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 , NR 6 S(O) 2 R 6 , NHC(O)OR 6 , NR 6 C(O)OR 6 , NHC(O)NH 2 ,
  • r R> 6 i • s r R> 7 , r R> 8 , r R> 9 , or o 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 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 ⁇ 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 ; 12
  • 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 16E , 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;
  • 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; and 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- ⁇ (3S)-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ pyrrolidine, one or more PEG-lipid conjugates, and one or more nucleic acids.
  • the Lipid- Based Particle comprises, cholesterol, DSPC, l- ⁇ (3S)-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ 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- ⁇ (3S)-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ 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- ⁇ (3S)-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ 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- ⁇ (3S)-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ 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- ⁇ (3S)-3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ 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- ⁇ (3R)-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ pyrrolidine, one or more PEG-lipid conjugates, and one or more nucleic acids.
  • the Lipid- Based Particle comprises, cholesterol, DSPC, l- ⁇ (3R)-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ 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- ⁇ (3R)-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ 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- ⁇ (3R)-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ 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- ⁇ (3R)-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ 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- ⁇ (3R)-3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ 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- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ -N,N-dimethylpyrrolidin-3-amine, one or more PEG-lipid conjugates, and one or more nucleic acids.
  • the Lipid- Based Particle comprises, cholesterol, DSPC, l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ -N,N-dimethylpyrrolidin-3-amine, 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- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ -N,N- dimethylpyrrolidin-3 -amine 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 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
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ -N,N-dimethylpyrrolidin-3-amine, 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- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ -N,N-dimethylpyrrolidin-3-amine
  • the PEG-lipid conjugate is N-(2,3- dimyristyloxypropyl)carbamate polyethyleneglycol-2000
  • 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 1 - ⁇ 3 ,4-bis[(9Z, 12Z)-octadeca-9, 12-dienyloxy]butyl ⁇ -N,N-dimethylpyrrolidin-3-amine 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- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ 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- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ 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 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
  • l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ pyrrolidine is about 5-60 weight/weight %
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ pyrrolidine, the PEG-lipid conjugate is 2-(octadecyloxy)-l- ((octadecyloxy)methyl)ethyl 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, 96,99,102,105,108,111,114,117,120,123,126,129,132,135,138- hexatetracontaoxanonatriacontahect-1-ylcarbamate, and the therapeutic agent is siRNA.
  • a further embodiment pertains to a Lipid-Based Particle, wherein the 2- (octadecyloxy)- 1 -((octadecyloxy)methyl)ethyl
  • 96,99,102,105,108,111,114,117,120,123,126,129,132,135,138- hexatetracontaoxanonatriacontahect-1-ylcarbamate 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- ⁇ 3,4- bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ 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- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ 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, and the l- ⁇ 3,4- bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ pyrrolidine is about
  • a further embodiment pertains to a Lipid-Based Particle, wherein the non-cationic lipids are cholesterol and DSPC, the cationic lipid is l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ pyrrolidine, the PEG-lipid conjugate is N-[3,4-bis(octadecyloxy)butyl]-
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-[3,4- bis(octadecyloxy)butyl] -
  • 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- ⁇ 3,4- bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ 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- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ pyrrolidine, the PEG-lipid conjugate is 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- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ pyrrolidine
  • the PEG-lipid conjugate is 1 ,2-dipalmitoyl-sn-glycerol- methoxypolyethyleneglycol-2000
  • 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- ⁇ 3,4-bis[(9Z,12Z)-octadeca- 9,12-dienyloxy]butyl ⁇ 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- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ pyrrolidine, the PEG-lipid conjugate is N-(carbonyl- methoxypolyethyleneglycol-2000)-l,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- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butyl ⁇ pyrrolidine
  • the PEG-lipid conjugate is N-(carbonyl- methoxypolyethyleneglycol-2000)-l,2-dipalmitoyl-sn-glycero-3-phospho
  • a further embodiment pertains to a Lipid-Based Particle, wherein the N-(carbonyl- methoxypolyethyleneglycol-2000)-l,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- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ 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.
  • 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 I, II, III, or IV 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 I, II, III, or IV 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- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ piperidine, 4- ⁇ 3,4-bis[(9Z,12Z)- octadeca-9,12-dienyloxy]butyl ⁇ morpholine, N,N-diethyl-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butan- 1 -amine, N,N-dimethyl-3 ,4-bis[(9Z, 12Z)-octadeca-9, 12-dienyloxy]butan- 1 - amine, l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butan- 1 -
  • functional CaBLES comprising one or more (PEG)- lipid conjugates, one or more non-cationic lipids, and one or more cationic lipids chosen from l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ piperidine, 4- ⁇ 3,4-bis[(9Z,12Z)- octadeca-9,12-dienyloxy]butyl ⁇ morpholine, N,N-diethyl-3,4-bis[(9Z,12Z)-octadeca-9,12- dienyloxy]butan- 1 -amine, N,N-dimethyl-3 ,4-bis[(9Z, 12Z)-octadeca-9, 12-dienyloxy]butan- 1 - amine, l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butan- 1 -
  • 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
  • a further embodiment pertains to examples of PEG-lipid conjugates that are useful for the practice of this invention which include, but are not limited to, 2-(tetradecyloxy)-l- ((tetradecyloxy)methyl)ethyl 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, 96,99,102,105,108,111,114,117,120,123,126,129,132,135,138- hexatetracontaoxanonatriacontahect- 1 -ylcarbamate, 2-(hexadecyloxy)- 1 - ((hexadecyloxy)methyl)ethyl
  • PEG-lipid conjugates are described in, e.g., US App. No. 61/095,748, which was filed on September 10, 2008 and is incorporated herein by reference.
  • 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)-l,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, l,2-dipalmitoyl-sn-glyce
  • 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)-l,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-methoxypolyethyleneglycol- 2000, and N-[3,4-bis(hexadecyloxy)butyl]-
  • 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- methoxypolyethyleneglycol-2000)- 1 ,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1 ,2-distearoyl-sn-glycerol-methoxypolyethyleneglycol-2000 and N-(carbonyl- methoxypolyethyleneglycol-2000)- 1 ,2-dimyristoyl-sn-glycero-3-phosphoethanolamine, 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
  • the cationic lipids of the CaBLES and Lipid-Based 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)
  • Y is CH 2 , NH or O
  • Y is a bond or C 1 -C 6 alky lene
  • R and R are independently H, cycloalkyl, cycloalkenyl or R ; or
  • R and R together with the nitrogen to which they are attached, are heterocycloalkyl or heteroaryl;
  • R and R is H, and the other is C 14 -C 2 o-alkenyl, or C 14 -C 2 o-alkyl; or
  • R and R are independently selected C 14 -C 2 o-alkenyl, or C 14 -C 2 o-alkyl; or 3 4 20 21 20 21
  • R and R together are CR R , wherein R is H and R is C 14 -C 2 o-alkenyl, C 14 - C 2 o-alkyl, or CH 2 O-C 14 -C 2 o-alkenyl; or R and R are independently selected C 14 -C 2 o-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 ,
  • 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 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 ⁇ 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 ⁇ NR 11 C(O)NHR 11 , NR 11 C(O)N(R 1 ⁇ 2 , C(O)NH 2 , C(O)NHR 11 , C(0)N(R U ) 2 , C(O)NH 2
  • R is R , R , R , or R ; 12
  • 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 16E , 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;
  • 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;
  • a further embodiment pertains to a method of making 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).
  • 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.
  • amount of 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.
  • 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 I, II, III, or IV, 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, emulsifiers, 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, emulsifiers, extenders, fillers, flavoring agents, humectants, lubricants, perfumes, preservatives, propellants, releasing agents, sterilizing agents, sweeteners, solubilizers, wetting agents, mixtures thereof and the
  • 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,
  • Excipients for preparation of compositions comprising a compound having formula (I) to be administered ophthalmically or orally include, but are not limited to, 1,3-butylene glycol, castor oil, corn oil, cottonseed oil, ethanol, fatty acid esters of sorbitan, germ oil, groundnut oil, glycerol, isopropanol, olive oil, polyethylene glycols, propylene glycol, sesame oil, water, mixtures thereof and the like.
  • Excipients for preparation of 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.
  • Combination Therapy 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 homo log (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
  • 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'-methoxy ethyl 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.
  • the term "multispecif ⁇ c 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.
  • DVDs may be monospecific, i.e., capable of binding one antigen or multispecif ⁇ c, 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.
  • Antimetabolites include ALIMT A ® (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-
  • 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- 202, R-roscovitine), ZK-304709 and the like.
  • 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)-l-(4- sulfamoylphenyl-lH-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,
  • 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.
  • TRAIL or death receptors e.g., pro- apoptotic receptor agonists DR4 and DR5
  • conatumumab e.g., pro- apoptotic receptor agonists DR4 and DR5
  • conatumumab e.g., ETR2-ST01
  • GDCO 145 e.g., ETR2-ST01
  • GDCO 145 e.g., GDCO 145
  • HGS-1029 e.g., 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-001, rapamycin, temsirolimus and the like.
  • Non-steroidal anti-inflammatory drugs include AMIGESIC (salsalate), DOLOBID (diflunisal), MOTRIN ® (ibuprofen), ORUDIS ® (ketoprofen), RELAFEN ® (nabumetone), FELDENE (piroxicam), ibuprofen cream, ALEVE (naproxen) and NAPROSYN (naproxen), VOLTAREN ® (diclofenac), INDOCIN ® (indomethacin), CLINORIL ® (sulindac), TOLECTIN ® (tolmetin), LODINE ® (etodolac), TORADOL ® (ketorolac), DAYPRO ® (oxaprozin) and the like.
  • 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.
  • Antibiotics include intercalating antibiotics aclarubicin, actinomycin D, amrubicin, annamycin, adriamycin, BLENOXANE (bleomycin), daunorubicin, CAELYX or
  • MYOCET liposomal doxorubicin
  • elsamitrucin epirbucin
  • glarbuicin glarbuicin
  • ZAVEDOS idarubicin
  • mitomycin C nemorubicin
  • neocarzinostatin peplomycin
  • pirarubicin rebeccamycin
  • stimalamer streptozocin
  • VALSTAR valrubicin
  • Topoisomerase inhibitors include aclarubicin, 9-aminocamptothecin, amonafide, amsacrine, becatecarin, belotecan, BN-80915, CAMPTOSAR (irinotecan hydrochloride), camptothecin, CARDIOXANE (dexrazoxine), diflomotecan, edotecarin, ELLENCE or PHARMORUBICIN (epirubicin), etoposide, exatecan, 10-hydroxycamptothecin, gimatecan, lurtotecan, mitoxantrone, orathecin, pirarbucin, pixantrone, rubitecan, sobuzoxane, SN-38, tafluposide, topotecan and the like.
  • aclarubicin 9-aminocamptothecin, amonafide, amsacrine, becatecarin, belotecan
  • Antibodies include AVASTIN (bevacizumab), CD40-specif ⁇ c antibodies, chTNT- 1/B, denosumab, ERBITUX ® (cetuximab), HUMAX-CD4 ® (zanolimumab), IGFlR-specific antibodies, lintuzumab, PANOREX ® (edrecolomab), RENCAREX ® (WX G250),
  • RITUXAN rituximab
  • ticilimumab ticilimumab
  • trastuzimab tuzimab
  • 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 ®
  • NOLVADEX ® tamoxifen citrate
  • PLENAXISTM abarelix
  • prednisone PROPECIA ®
  • 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.
  • agents include ALF AFERONE ,(IFN- ⁇ ), BAM- 002 (oxidized glutathione), BEROMUN ® (tasonermin), BEXXAR ® (tositumomab), CAMPATH (alemtuzumab), CTLA4 (cytotoxic lymphocyte antigen 4), decarbazine, denileukin, epratuzumab, GRANOCYTE (lenograstim), lentinan, leukocyte alpha interferon, imiquimod, MDX-OlO (anti-CTLA-4), melanoma vaccine, mitumomab, molgramostim, MYLOTARGTM (gemtuzumab ozogamicin), NEUPOGEN (filgrastim), Onco VAC-CL, OVAREX ® (oregovomab), pemtumomab (Y-muHMFGl), PROVENGE ®
  • 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 to direct them to have anti-tumor activity and include include krestin, lentinan, sizofiran, 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 (gemcitabine), TOMUDEX ® (ratitrexed), TROXATYLTM (triacetyluridine troxacitabine) and the like.
  • Purine analogs include LANVIS ® (thioguanine) and PURI-NETHOL ®
  • 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, II, III, or IV 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-l,4- diene), AVAGE (tazarotene), AVE-8062 (combreastatin derivative) BEC2 (mitumomab), cachectin or cachexin (tumor necrosis factor), canvaxin (vaccine), CEAVAC (cancer vaccine), CELEUK ® (celmol
  • PANVAC -VF investment cancer vaccine
  • pegaspargase pegaspargase
  • PEG Interferon A PEG Interferon A
  • CR CR) CR) phenoxodiol, procarbazine, rebimastat, REMOVAB (catumaxomab), REVLIMID (lenalidomide), RSRl 3 (efaproxiral), SOMATULINE ® LA (lanreotide), SORIAT ANE ®
  • CR CR
  • necrosis factor- ⁇ TRACLEER or ZAVESCA (bosentan), tretinoin (Retin-A), tetrandrine, TRISENOX (arsenic trioxide), VIRULIZIN , ukrain (derivative of alkaloids from the
  • CaBLES comprise one or more non-cationic lipids, one or more cationic lipids having Formula I, II, III, or IV 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 J. 20: 6877-6888 (2001) are combined with rational design rules set forth in Reynolds et al., Nature Biotech. 22(3):326-330 (2004)). Further enhancing, isolating, synthesizing and generating of the siRNA can be done by various methods known in the art, (see, e.g., Elbashir, et al., EMBO J. 20: 6877-6888 (2001); Elbashir, et al., Genes Dev.
  • 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.
  • 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, II, III, or IV. 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, l,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, l,2-dioleoyl-3-trimethylammonium-propane chloride, l,2-dilineoyl-3-dimethylammoni
  • Lipid-Based Particles are a mixture of one or more cationic lipids of Formula I, II, III, or IV (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. Ribogreen Assay for Measuring SiRNA Concentration and Encapsulation Efficiency of of of
  • 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-IOO, or phenol/chloroform.
  • 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 0 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.). Plates were read at 20 0 C with an acquisition time of 50 seconds for each sample, and data was analyzed with Wyatt Technology's Dynamics V6 software. To rule out potential multiple scattering artifacts, a second plate at 4-fold reduced sample concentrations was independently prepared by mixing 5 ⁇ L stock solutions with 95 ⁇ L PBS. Under our experimental conditions the results at the two concentrations were very similar, and the final reported result for each sample represents the average of values obtained from the two plates.
  • 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.U SEQ ID NO: 1) (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 0 C. Scattered light was detected at a 173° backward scattering angle. The viscosity and refractive index of water at 25 0 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.
  • DMEM Dulbecco's Modified Eagles Medium, Invitrogen Corp.
  • 10% fetal bovine serum Invitrogen Corp.
  • Formulated or unformulated siRNAs were administrated via tail vein (i.v) injection.
  • IHC was carried out as previously described [Li, L., et al, Evaluating hypoxia- inducible factor- 1 alpha as a cancer therapeutic target via inducible RNA interference in vivo. Cancer Res, 2005. 65(16): p. 7249-58]. Briefly, tumors were excised, cut into pieces of less than 3 mm in thickness and immediately fixed in buffered formalin solution with neutral pH (Sigma, St. Louis, MO). The formalin-fixed and paraffin-embedded tumor sections were then used for staining. The mouse anti- ⁇ -galactosidase mAb (Promega, Madison, WI) was used to detect ⁇ -galactosidase in tumor sections.
  • DAB (3,3'-diaminobenzidine) was used as the chromogen.
  • IHC 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 +/-, there was 5% of the section with weak staining or one patch of strong staining +, there was 15% of the section with strong staining ++, there was ⁇ 50% of the section with strong straining , there was 80% of the section with strong straining +, the whole section had strong 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
  • 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 l,8-diazabicyclo(5.4.0)undec- 7-ene;
  • DCC means dicyclohexylcarbodiimide;
  • DIBAL means diisobutylaluminum hydride;
  • DIEA means diisopropylethylamine;
  • DMAP means N,N-dimethylaminopyridine;
  • 2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethyl 4- methylbenzenesulfonate which can be prepared as described in Example IA, when reacted with an amine of Formula (1) wherein R 1 and R 2 are as described herein, with a base such as but not limited to N,N-diisopropylethylamine, will provide a compound of Formula (2).
  • the reaction may be conducted in a commercial single mode microwave at elevated temperature.
  • Anhydrous solvents such as but not limited to 1,4-dioxane are typically employed.
  • compounds of Formula (3) can be prepared from compounds of Formula (2) by hydrolysis of the former using an acid such as but not limited to hydrochloric acid.
  • the reaction can be performed immediately after the formation of (2) in the same reaction vessel.
  • Compounds of Formula (I), wherein R 1 , R 2 , R 3 , and R 4 are described herein, can be prepared by reacting compounds of Formula (3) with base, such as but not limited to sodium hydride, with a compound of Formula (4).
  • the reaction is typically performed at an elevated temperature in a solvent such as but not limited to toluene.
  • R 3 and R 4 can be the same, two equivalents of (4) can be used. IfR 3 and R 4 are to be different, one equivalent of (4) 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 Y 3 and Y 4 are carbonyl can be prepared from compounds of Formula (3) by coupling with an acid of Formula (5) under standard coupling conditions known in the art and widely available in the literature. If it is desired for R 3 and R 4 to be the same, two equivalents of (8) can be used. IfR 3 and R 4 are to be different, one equivalent of (8) can be used to obtain a compound wherein R 4 is H after purification. This intermediate can then be reacted with R 4 COOH to obtain a compound of Formula (I).
  • 2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethyl 4- methylbenzenesulfonate which can be prepared as described in Example IA, when reacted with an amino alcohol of Formula (6), wherein R 1 , R 2 , and Y 1 are as described herein, with a base such as but not limited to sodium hydride, will provide a compound of Formula (7).
  • Compounds of Formula (8) can be prepared from compounds of Formula (7) by hydrolysis of the latter using an acid such as aqueous hydrochloric acid in a solvent such as but not limited to tetrahydrofuran.
  • compounds of Formula (9) can be prepared from 2-(2,2- dimethyl- l,3-dioxolan-4-yl)ethanol using chloromethyl methyl ether and a base such as but not limited to N-ethyl-N-isopropylpropan-2-amine in a solvent such as but not limited to dichloromethane.
  • Compounds of Formula (10) can be prepared from compounds of Formula (9) by hydrolysis of the latter using an acid such as but not limited to acetic acid in a solvent such as water.
  • Compounds of Formula (11), wherein Y 3 and Y 4 are a bond wherein R 3 and R 4 are described herein, can be prepared by reacting compounds of Formula (10) with base, such as but not limited to sodium hydride, with a compound of Formula (4). The reaction is typically performed 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 (4) can be used. IfR 3 and R 4 are to be different, one equivalent of (4) 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).
  • base such as but not limited to sodium hydride
  • Compounds of Formula (11), wherein Y 3 and Y 4 are carbonyl can be prepared from compounds of Formula (10) by coupling with an acid of Formula (5) under standard coupling conditions known in the art and widely available in the literature. If it is desired for R 3 and R 4 to be the same, two equivalents of (8) can be used. IfR 3 and R 4 are to be different, one equivalent of (8) can be used to obtain a compound wherein R 4 is H after purification. This intermediate can then be reacted with R 4 COOH to obtain a compound of Formula (III).
  • Compounds of Formula (12) can be prepared from compounds of Formula (11) by hydrolysis of the latter using an acid such as but not limited to hydrochloric acid.
  • Compounds of Formula (13) can be prepared from compounds of Formula (12) by oxidizing the latter using an oxidant such as but not limited to Dess-Martin Periodinane or any other oxidant known in the art or available in the literature.
  • an oxidant such as but not limited to Dess-Martin Periodinane or any other oxidant known in the art or available in the literature.
  • Compounds of Formula (14) can be prepared from compounds of Formula (13) by using a Lindgren oxidation or some other suitable oxidation procedure known in the art and available in the literature.
  • Compounds of Formula (III) can be prepared by reacting compounds of Formula (14) with a compound of Formula (15) where R 1 , R 2 and Y 6 are defined herein, using standard coupling conditions known in the art and widely available in the literature.
  • compounds of Formula (15) can be prepared from compounds of Formula (12), wherein Y 3 , Y 4 , R 3 and R 4 are defined herein, using reagents such as DPPA (diphenylphosphoryl azide) and DEAD (diethyl azodicarboxylate) in a solvent such as but not limited to tetrahydrofuran.
  • reagents such as DPPA (diphenylphosphoryl azide) and DEAD (diethyl azodicarboxylate) in a solvent such as but not limited to tetrahydrofuran.
  • Compounds of Formula (16) can be prepared from compounds of Formula (15) by using a reagent such as triphenylphosphine in solvents such as but not limited to water and tetrahydrofuran.
  • Compounds of Formula (IV) can be prepared from compounds of Formula (16) by coupling of an acid of Formula (17) wherein R 1 , R 2 and Y 5 are described herein, under standard coupling conditions known
  • a base such as but not limited to N-ethyl-N-isopropylpropan- 2-amine in a solvent such as dichloromethane.
  • Compounds of Formula (IV), wherein Y 2 is O and Y 5 is NH, can be prepared from compounds of Formula (20) by coupling with an amine of Formula (21), wherein R 1 , R 2 and Y 6 are described herein, in the presence of a base such as but not limited to N-methyl morpholine in a solvent such as but not limited to dichloromethane .
  • Example IB l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ piperidine 2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethyl 4-methylbenzenesulfonate (Example IA, 500 mg) was placed in a microwave reaction vial along with piperidine (1-2 eq.), Hunig's base (2 eq) and dioxane (2.2 ml). The reaction was placed in the microwave (Biotage Initiator) for 15 minutes at 140 0 C.
  • Example 2 4- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ morpholine
  • Example 2 was prepared using the procedure described in Example IB, substituting morpholine for piperidine.
  • MS (ESI) m/z 612.6 (M+H) + ; 1 U NMR (300 MHz, CDCl 3 ) ⁇ 5.26
  • Example 3 was prepared using the procedure described in Example IB, substituting diethyl amine for piperidine.
  • Example 4 was prepared using the procedure described in Example IB, substituting dimethyl amine hydrochloride for piperidine.
  • Example 5 was prepared using the procedure described in Example IB, substituting 4- phenylpiperazine for piperidine.
  • Example 6 l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ -4-methylpiperazine
  • Example 6 was prepared using the procedure described in Example IB, substituting A- methylpiperazine for piperidine.
  • MS (ESI) m/z 685.6 (M+H) + ; 1 U NMR (300 MHz, CDCl 3 ) ⁇ 5.23 - 5.46 (m, 8 H) 3.50 - 3.66 (m, 1 H) 3.33 - 3.50 (m, 6 H) 2.72 - 2.82 (m, 4 H) 2.36 -
  • Example 7 was prepared using the same procedure described in Example IB, substituting 2-methoxy-N-methylethanamine for piperidine.
  • Example 8 1 - ⁇ 3 ,4-bis[(9Z, 12Z)-octadeca-9, 12-dienyloxy]butyl ⁇ -4-(2-methoxyphenyl)piperazine
  • Example 8 was prepared using the same procedure described in Example IB, substituting l-(2-methoxyphenyl)piperazine for piperidine.
  • Example 9 was prepared using the same procedure in Example IB, substituting V,V,N 2 -trimethylethane-l,2-diamine for piperidine.
  • Example 10 N- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,
  • Example 10 was prepared using the same procedure in Example IB, substituting N- methyl-2-(pyridin-2-yl)ethanamine for piperidine. MS (ESI) m/z 121.6 (M+H) + ; 1 H NMR
  • Example 11 was prepared using the same procedure in Example IB, substituting N- methyl- 1-phenylmethanamine for piperidine. MS (ESI) m/z 706.6 (M+H) + ; 1 H NMR (300)
  • Example 12 was prepared using the same procedure in Example IB, substituting l-(4- fluorophenyl)-N-methylmethanamine for piperidine.
  • Example 13 l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ -4-(2-fluorophenyl)piperazine
  • Example 13 was prepared using the same procedure in Example IB, substituting l-(2- fluorophenyl)piperazine for piperidine.
  • Example 14 was prepared using the same procedure in Example IB, substituting N- benzylethanamine for piperidine.
  • Example 15 N- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ -N-ethyl-N',N'-dimethylethane-l,2- diamine
  • Example 15 was prepared using the same procedure in Example IB, substituting N 1 - ethyl-N 2 ,N 2 -dimethylethane-l,2-diamine for piperidine.
  • Example 16 1 - ⁇ 3 ,4-bis[(9Z, 12Z)-octadeca-9, 12-dienyloxy]butyl ⁇ -N,N-dimethylpiperidin-4-amine
  • Example 16 was prepared using the same procedure in Example IB, substituting N,N-dimethylpiperidin-4-amine for piperidine.
  • Example 17 was prepared using the same procedure in Example IB, substituting N,N-dimethylpyrrolidin-3 -amine for piperidine.
  • Example 18 was prepared using the same procedure in Example IB, substituting bis(2-methoxyethyl)amine for piperidine.
  • Example 19 l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ -4-methoxypiperidine
  • Example 18 was prepared using the same procedure in Example IB, substituting 4- methoxypiperidine for piperidine.
  • Example 2OB l- ⁇ (3R)-3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ pyrrolidine
  • Example 2OB was prepared using the same procedure in Example IB, substituting Example 2OA for Example IA and pyrrolidine for piperidine.
  • MS (ESI) m/z 656.6 (M+H) + ;
  • 1 H NMR 300 MHz, CDCl 3 ) ⁇ 5.25 - 5.45 (m, 8 H) 3.51 - 3.63 (m, 1 H) 3.33 - 3.51 (m, 6 H)
  • Example 2 IB l- ⁇ (3S)-3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ pyrrolidine
  • Example 2 IB was prepared using the same procedure in Example IB, substituting Example 21 A for Example IA and pyrrolidine for piperidine.
  • Example 22 was prepared using the same procedure in Example IB, substituting Example 2OA for Example IA and diethyl amine for piperidine.
  • Example 23 was prepared using the same procedure in Example IB, substituting Example 21A for Example IA and diethyl amine for piperidine.
  • Example 24 l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ pyrrolidine
  • Example 24 was prepared using the same procedure in Example IB, substituting pyrrolidine for piperidine.
  • Example 25 was prepared as described in Example 31 using the appropriate reagents and conditions.
  • Example 26 was prepared as described in Example 31 using the appropriate reagents and conditions.
  • Example 27 l-(2- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butoxy ⁇ ethyl)aziridine
  • Example 27 was prepared as described in Example 31 using the appropriate reagents and conditions.
  • Example 28 was prepared as described in Example 31 using the appropriate reagents and conditions.
  • MS (ESI) m/z 729.6 (M+H); 1 U NMR (400 MHz, CHLOROFORM-D) ⁇ ppm 5.26 - 5.44 (m, 8 H) 3.46 - 3.63 (m, 6 H) 3.35 - 3.47 (m, 5 H) 2.73 - 2.81 (m, 4 H) 2.58 (t,
  • Example 30 was prepared as described in Example 31 using the appropriate reagents and conditions.
  • Example 30A 4-(diethylamino)butane- 1 ,2-diol
  • Example IA 500 mg, 1.66 mmol
  • diethylamine 609 mg, 8.32 mmol
  • Example 30A (335 mg, 1.66 mmol) was dissolved in tetrahydrofuran and 2N HCl (aq) (8.5 mL each) and stirred overnight at room temperature. The acidic solution was made basic by the addition of 5 N NaOH ( aq ). The aqueous layer was extracted with chloroform (6x) and the combined organics were dried (MgSO 4 ), filtered and concentrated by rotary evaporation. The residue was used in the next step without further purification.
  • Example 30 The reaction was stirred overnight at room temperature. The reaction was quenched with water and diluted with ethyl acetate. The aqueous layer was extracted with ethyl acetate (3x). The combined organics were dried (Na2SC>4), filtered and concentrated by rotary evaporation. The residue was purified by regular phase chromatography (Analogix, 0-100% hexanes: ethyl acetate) to give Example 30.
  • Example 31A 1 -(2-(2-(2,2-dimethyl- 1 ,3-dioxolan-4-yl)ethoxy)ethyl)pyrrolidine
  • Example 3 IA The aqueous layer was extracted (2x) with ethyl acetate and the combined organics were dried (Na 2 SO 4 ), filtered and concentrated. The residue was purified by regular phase chromatography (Analogix, 0 - 100% hexanes:ethyl acetate) to give Example 3 IA.
  • Example 3 IB l-(2- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butoxy ⁇ ethyl)pyrrolidine
  • Example 31A (423 mg, 1.7 mmol) was dissolved in tetrahydrofuran (1-2 mL) and 4N HCl aq (10 mL) and the reaction was stirred overnight at room temperature. The acidic solution was basified with 6N NaOH (aq) and extracted with chloroform (6x).
  • Example 31 The aqueous layer was extracted with ethyl acetate (3x) and the combined organics were dried (Na 2 SO 4 ), filtered and concentrated by rotary evaporation. The residue was purified by regular phase chromatography (Analogix 0-100% hexanes: ethyl acetate) to give Example 31.
  • Example 30A 25 mg, 0.15 mmol
  • (5Z,8Z,26Z,29Z)-pentatriaconta-5,8,26,29- tetraen-18-one (93 mg, 0.19 mmol) were combined in toluene (3 mL).
  • /?-Toluenesulfonic acid (2.95 mg, 0.016 mmol) was added, the flask was fitted with a small Dean-Stark trap and the reaction was refluxed overnight.
  • Triethylamine, ethyl acetate and water were added and the layers were separated.
  • Example 32 The aqueous layer was extracted with ethyl acetate (2x) and the combined organics were dried (Na 2 SO 4 ), filtered and concentrated by rotary evaporation. The residue was purified by regular phase chromatography (Analogix, 0 - 100% hexanes:ethyl acetate) to give Example 32.
  • Example 33 A was prepared using the procedure from Example 3OA, substituting pyrrolidine for diethylamine.
  • MS (ESI) m/z 199.9 (M+H); 1 U NMR (300 MHz, CHLOROFORM-D) ⁇ ppm 4.02 - 4.20 (m, 2 H) 3.49 - 3.59 (m, 1 H) 2.39 - 2.65 (m, 6 H) 1.66 - 1.94 (m, 6 H) 1.41 (s, 3 H) 1.36 (s, 3 H).
  • Example 33B was prepared from Example 33A using the procedure from Example 3OB.
  • 1 H NMR 300 MHz, CHLOROFORM-D
  • ppm 3.84 - 3.96 m, 1 H) 3.56 - 3.65 (m, 1 H) 3.46 - 3.56 (m, 1 H) 2.88 - 3.01 (m, 1 H) 2.59 - 2.72 (m, 1 H) 2.45 - 2.59 (m, 4 H) 1.70 - 1.85 (m, 6 H).
  • Example 33B (300 mg, 1.9 mmol) was dissolved in dichloromethane (9.4 ml). Oleic acid (1.17 g, 4.15 mmol) was added followed by l-ethyl-3-[3-(dimethylamino)propyl]- carbodiimide hydrochloride (903 mg, 4.71 mmol) and 4-dimethylaminopyridine (46 mg, 0.38 mmol). The reaction was stirred overnight at room temperature. The reaction was quenched with water and diluted with ethyl acetate. The aqueous layer was extracted with ethyl acetate (3x) and the combined organics were dried (Na2SO4), filtered and concentrated by rotary evaporation.
  • Example 33B (400 mg, 2.51 mmol) was dissolved in toluene (7 mL). Sodium hydride (90%, 482 mg, 20.1 mmol) was added portion- wise. The flask was flushed with nitrogen and stirred for 30 minutes. (Z)-octadec-9-enyl methanesulfonate (2.1 g, 6.0 mmol) was taken up in toluene (3 mL) and added dropwise to the mixture. The flask was fit with a Vigourex column and refluxed for 4 hours. The reaction was cooled, quenched with ethanol and water and diluted with ethyl acetate.
  • Example 34 MS (ESI) m/z 660.7 (M+H); 1 H NMR (300 MHz, CHLOROFORM-D) ⁇ ppm 5.27 - 5.42 (m, 4 H) 3.50 - 3.62 (m, 1 H) 3.35 - 3.49
  • Example 35 was prepared using the same procedure as Example 33, substituting (5Z,8Z,1 lZ,14Z)-icosa-5,8,l 1,14-tetraenoic acid for oleic acid.
  • Example 36A (S)-4-(2-(methoxymethoxy)ethyl)-2,2-dimethyl- 1 ,3-dioxolane
  • Hunig's base (15.1 ml, 87 mmol)
  • the reaction was cooled to O 0 C and chloro(methoxy)methane (3.1 ml, 40.4 mmol) was added dropwise.
  • the reaction was warmed to room temperature and stirred overnight.
  • the solution was diluted with ether and poured into saturated NH4Cl( aq ).
  • Example 36A The aqueous layer was extracted with diethyl ether (2x) and the combined organics were dried (Na 2 SO 4 ), filtered and concentrated by rotary evaporation. The residue was purified by regular phase chromatography (Analogix, 0- 100%, hexanes : ethyl acetate) to give Example 36A.
  • Example 36A was taken up in water (29 ml) and acetic acid (44 ml) and stirred for 2 hours at room temperature, monitoring very carefully by TLC to avoid deprotection of the
  • MOM methoxymethyl
  • Example 36C (6Z,9Z)-18-((S)-4-(methoxymethoxy)-l-((9Z,12Z)-octadeca-9,12-dienyloxy)butan-2- yloxy)octadeca-6,9-diene
  • Example 36D (S)-3,4-bis((9Z,12Z)-octadeca-9,12-dienyloxy)butan-l-ol
  • Example 36C (1.84 g, 2.84 mmol) was dissolved in tetrahydrofuran (10.6 ml) and methanol (3.55 ml). To this solution, concentrated HCl( aq ) (5 ml) was added. The reaction was heated to 62 0 C for 5 hours, cooled to room temperature and 6 N NaOH was added until the solution was basic. The aqueous layer was extracted with ethyl acetate (3x) and the combined organics were dried (Na 2 SO 4 ), filtered and concentrated by rotary evaporation.
  • Example 36D (S)-3,4-bis((9Z, 12Z)-octadeca-9, 12-dienyloxy)butyl 4-nitrophenyl carbonate
  • Example 36D (27 mg, 0.045 mmol) was taken up in dichloromethane (1 ml) and triethylamine (6.2 uL) was added followed by 4-nitrophenyl carbonochloridate (11 mg, 0.054 mmol). The reaction was allowed to stir overnight. The reaction mixture was loaded directly onto a silica gel column and purified (Analogix, 0-75%, hexanes : ethyl acetate) to provide Example 36E.
  • Example 36F (3 S)-3 ,4-bis [(9Z, 12Z)-octadeca-9, 12-dienyloxy]butyl 3 -pyrrolidin- 1 -ylpropylcarbamate
  • Example 36E (34.4 mg, 0.045 mmol) and 3-(pyrrolidin-l-yl)propan-l-amine (excess) were combined in dichloromethane (1 ml) and stirred overnight. The mixture was loaded directly onto silica gel for purification (Analogix, 0-100%, hexanes : ethyl acetate) to give Example 36.
  • Example 37 was prepared using the same procedure described in Example IB substituting pyrrolidine for piperidine and octadecyl methanesulfonate for (9Z,12Z)- octadeca-9,12-dienyl methanesulfonate.
  • Example 38 was prepared using the same procedure described in Example IB, substituting pyrrolidine for piperidine and hexyldecyl methanesulfonate for (9Z,12Z)- octadeca-9,12-dienyl methanesulfonate.
  • Example 39 was prepared using the same procedure described in Example IB substituting pyrrolidine for piperidine and (E)-hexadec-9-enyl methanesulfonate for (9Z,12Z)-octadeca-9,12-dienyl methanesulfonate.
  • Example 40 was prepared using the same procedure described in Example IB substituting pyrrolidine for piperidine and (E)-octadec-9-enyl methanesulfonate for (9Z,12Z)- octadeca-9,12-dienyl methanesulfonate.
  • Example 41 l- ⁇ 3,4-bis[(9E,12E)-octadeca-9,12-dienyloxy]butyl ⁇ pyrrolidine
  • Example 41 was prepared using the same procedure described in Example IB substituting pyrrolidine for piperidine and (9E,12E)-octadeca-9,12-dienyl methanesulfonate for (9Z,12Z)-octadeca-9,12-dienyl methanesulfonate.
  • Example 42 1 - ⁇ 3 ,4-bis[(9Z, 12Z, 15Z)-octadeca-9, 12, 15 -trienyloxy]butyl ⁇ pyrrolidine
  • Example 42 was prepared using the same procedure described in Example IB substituting pyrrolidine for piperidine and (9Z,12Z,15Z)-octadeca-9,12,15-trienyl methanesulfonate for (9Z,12Z)-octadeca-9,12-dienyl methanesulfonate.
  • Example 44A 4-(2-(methoxymethoxy)ethyl)-2,2-dimethyl- 1 ,3-dioxolane
  • Example 44 A was prepared using the same procedure described for Example 36 A, substituting 2-(2,2-dimethyl-l,3-dioxolan-4-yl)ethanol for (S)-2-(2,2-dimethyl-l,3-dioxolan- 4-yl)ethanol.
  • Example 44B was prepared using the same procedure described for 36B, substituting Example 44 A for Example 36A and used without further purification.
  • Example 44C (6Z,9Z)-18-(4-(methoxymethoxy)-l-((9Z,12Z)-octadeca-9,12-dienyloxy)butan-2- yloxy)octadeca-6,9-diene
  • Example 44C was prepared using the same procedure described for 36C, substituting
  • Example 44D was prepared using the same procedure described for 36D, substituting Example 44C for Example 36C.
  • Example 45 was prepared using the procedure described for Example 44, substituting N ⁇ N ⁇ N 3 -trimethylpropane-l,3-diamine for 3-(lH-imidazol-l-yl)propan-l-amine.
  • MS (ESI) m/z 701.6 (M+H); 1 H NMR (300 MHz, CHLOROFORM-D) ⁇ ppm 5.24 - 5.47 (m, 8 H) 3.51
  • Example 46 was prepared using the procedure described for Example 44, substituting l-(pyrrolidin-3-yl)-lH-imidazole for 3-(lH-imidazol-l-yl)propan-l-amine.
  • MS (ESI) m/z 722.6 (M+H); 1 H NMR (300 MHz, CHLOROFORM-D) ⁇ ppm 7.54 - 7.68 (m, 1 H) 7.07 - 7.14 (m, 1 H) 7.04 (s, 1 H) 5.25 - 5.45 (m, 8 H) 4.58 - 4.71 (m, 1 H) 3.54 - 3.69 (m, 1 H) 3.33 - 3.54 (m, 6 H) 2.80 - 3.07 (m, 2 H) 2.71 - 2.82 (m, 4 H) 2.36 - 2.72 (m, 4 H) 1.99 - 2.11 (m, 8 H) 1.64 - 2.01 (m, 4 H) 1.48 - 1.63 (m, 4 H) 1.22 - 1.
  • Example 47 was prepared using the procedure described for Example 44, substituting 3-(pyrrolidin-l-yl)propan-l-amine for 3-(lH-imidazol-l-yl)propan-l-amine.
  • MS (ESI) m/z 713.7 (M+H); 1 H NMR (300 MHz, CHLOROFORM-D) ⁇ ppm 5.25 - 5.45 (m, 8 H) 3.53 -
  • Example 48 was prepared using the procedure described for Example 44, substituting N ⁇ N ⁇ dimethylpropane-l ⁇ -diamine for 3-(lH-imidazol-l-yl)propan-l-amine. MS (ESI) m/z
  • Example 49 was prepared using the procedure described for Example 44, substituting azetidine for 3-(lH-imidazol-l-yl)propan-l-amine.
  • Example 50 l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ -2-methylpyrrolidine
  • Example 50 was prepared using the procedure described for Example 44, substituting 2-methylpyrrolidine for 3-(lH-imidazol-l-yl)propan-l-amine.
  • Example 51 l- ⁇ 3,4-bis[(9Z,12Z)-octadeca-9,12-dienyloxy]butyl ⁇ -2,5-dimethylpyrrolidine
  • Example 51 was prepared using the procedure described for Example 44, substituting 2,5-dimethylpyrrolidine for 3-(lH-imidazol-l-yl)propan-l-amine.
  • 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 52A (1.0 g) and dibenzylamine (0.657 mg) were placed in a microwave vial (Biotage) and dioxane (2.5 mL) was added. The vial was capped and placed in a microwave reactor (Biotage Initiator), and the mixture was heated at 15O 0 C for 30 minutes. The mixture was diluted with ethyl acetate and poured into water. The aqueous layer was extracted twice with ethyl acetate, and the extract was washed with brine, dried (Na 2 SO 4 ), filtered and concentrated. The concentrate was used in the next step without further purification.
  • Example 52B was added to tetrahydrofuran (20 mL) 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 52C 4-(dibenzylamino)butane- 1 ,2-diyl ditetradecanoate
  • tetradecanoic acid (1.68 g)
  • N 1 - ((ethylimino)methylene)-N 3 ,N 3 -dimethylpropane-l,3-diamine hydrochloride (1.41 g)
  • A- (dimethylamino)pyridine 45 mg
  • dichloromethane 5 mL
  • Water was added along with some brine and the aqueous layer was extracted with dichloromethane (3x).
  • the extract was dried (Na 2 SO 4 ), filtered and the filtrate was concentrated.
  • the concentrate was purified by flash column chromatography (Analogix 280, 0-50% ethyl acetate/hexanes) to provide the title compound.
  • Example 52D 500 mg was added to methanol/dichloromethane/ethyl acetate (1/1/1, 10 mL) 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 52C (1 g) in toluene (6 mL) and added to NaH (0.336 g, dry, 95%) in toluene (6 mL). The mixture was stirred at room temperature for 1 hour. Tetradecyl methanesulfonate (2.15 g) was added. The mixture was heated to 9O 0 C overnight. The mixture was cooled to room temperature and ethanol was added followed by water until the excess NaH was destroyed. The mixture was poured into water and brine and extreaced with ethyl acetate. The water was extracted with ethyl acetate, and the extract was dried (Na 2 SO 4 ), filtered and concentrated.
  • Example 53B was prepared using the procedure described for Example 52E, substituting Example 53A for Example 52D.
  • Example 53C was prepared using the procedure described for Example 52F, substituting Example 53B for Example 52E.
  • Example 54A was prepared using the procedure described for Example 53A, 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 54B was prepared using the procedure described for Example 53B, substituting Example 54A for Example 53A.
  • Example 54C was prepared using the procedure described for Example 52F, substituting Example 54B for Example 52E.
  • Example 55 A was prepared using the same procedure described for Example 53 A, substituting octadecyl methanesulfonate for tetradecyl methanesulfonate.
  • LCMS APCI
  • Example 55B was prepared using the same procedure described for Example 52E, substituting Example 55 A for Example 52D.
  • LCMS (APCI) m/z 610.9; 1 U NMR (300 MHz, CDCl 3 ) ⁇ 3.08-3.70 (m, 9H) 1.85-2.15 (m, 2H) 1.55 (s, 4H) 1.15-1.37 (m, 60H) 0.84-0.92 (m, 6H).
  • Example 55C was prepared using the same procedure described for Example 52F, substituting Example 55B for Example 52E.
  • Example 53B (100 mg) was dissolved in dichloromethane (1-2 mL) and mPEG-NPC (26.0 mg) was added. Hunig's base (26 mg) was added, and the mixture was stirred overnight at room temperature. The mixture was loaded directly onto a silica gel column (4 g Analogix) and chromatographed (Analogix 280, dichloromethane/methanol, 0-20%) to give Example 56.
  • Example 57 was prepared using the same procedure described for Example 56, substituting Example 54B for Example 53B.
  • Example 58 was prepared using the same procedure described for Example 56, substituting Example 55B for Example 53B.
  • Example 59 was prepared using the same procedure described for Example 52F, 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 60 was prepared using the same procedure described for Example 52F, 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, IH) 4.17-4.26 (m, 3H) 4.01-4.11 (m, IH) 3.83-3.91 (m, IH) 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).

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Abstract

L’invention concerne des lipides cationiques, des systèmes de libération de médicaments à base de lipides cationiques, des procédés de fabrication associés, ainsi que des méthodes de traitement de maladies au moyen desdits lipides.
PCT/US2009/040803 2008-04-16 2009-04-16 Lipides cationiques et utilisations associees WO2009129387A2 (fr)

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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
WO2011000106A1 (fr) * 2009-07-01 2011-01-06 Protiva Biotherapeutics, Inc. Lipides cationiques et procédés améliorés pour l'administration d'agents thérapeutiques
WO2012108397A1 (fr) * 2011-02-08 2012-08-16 第一三共株式会社 Nouveau lipide
WO2013135360A1 (fr) * 2012-03-16 2013-09-19 Merck Patent Gmbh Lipides d'acides aminés
JP2014526437A (ja) * 2011-06-07 2014-10-06 インツェラ・ゲゼルシャフト・ミト・ベシュレンクテル・ハフツング アミノ脂質、それらの合成及び使用
CN104168888A (zh) * 2012-03-16 2014-11-26 默克专利股份有限公司 靶向氨基酸脂质
US9018187B2 (en) 2009-07-01 2015-04-28 Protiva Biotherapeutics, Inc. Cationic lipids and methods for the delivery of therapeutic agents
US9181545B2 (en) 2004-06-07 2015-11-10 Protiva Biotherapeutics, Inc. Lipid encapsulating interfering RNA
US9301923B2 (en) 2009-12-23 2016-04-05 Novartis Ag Lipids, lipid compositions, and methods of using them
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
US10111871B2 (en) 2013-08-27 2018-10-30 Northeastern University Nanoparticle drug delivery system and method of treating cancer and neurotrauma
US10342761B2 (en) 2014-07-16 2019-07-09 Novartis Ag Method of encapsulating a nucleic acid in a lipid nanoparticle host
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
CN116535381A (zh) * 2023-07-06 2023-08-04 北京悦康科创医药科技股份有限公司 具有五元环缩醛结构的阳离子脂质化合物、包含其的组合物及用途

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US9926560B2 (en) 2004-06-07 2018-03-27 Protiva Biotherapeutics, Inc. Lipid encapsulating interfering RNA
US9181545B2 (en) 2004-06-07 2015-11-10 Protiva Biotherapeutics, Inc. Lipid encapsulating interfering RNA
WO2010030730A1 (fr) * 2008-09-10 2010-03-18 Abbott Laboratories Conjugués polyéthylène glycol-lipide et leurs utilisations
WO2010030739A1 (fr) * 2008-09-10 2010-03-18 Abbott Laboratories Conjugués lipidiques du polyéthylène glycol et leurs utilisations
US9018187B2 (en) 2009-07-01 2015-04-28 Protiva Biotherapeutics, Inc. Cationic lipids and methods for the delivery of therapeutic agents
WO2011000106A1 (fr) * 2009-07-01 2011-01-06 Protiva Biotherapeutics, Inc. Lipides cationiques et procédés améliorés pour l'administration d'agents thérapeutiques
US8569256B2 (en) 2009-07-01 2013-10-29 Protiva Biotherapeutics, Inc. Cationic lipids and methods for the delivery of therapeutic agents
US9301923B2 (en) 2009-12-23 2016-04-05 Novartis Ag Lipids, lipid compositions, and methods of using them
WO2012108397A1 (fr) * 2011-02-08 2012-08-16 第一三共株式会社 Nouveau lipide
JP2014526437A (ja) * 2011-06-07 2014-10-06 インツェラ・ゲゼルシャフト・ミト・ベシュレンクテル・ハフツング アミノ脂質、それらの合成及び使用
AU2013231819B2 (en) * 2012-03-16 2017-07-20 Merck Patent Gmbh Aminoacid lipids
RU2670618C2 (ru) * 2012-03-16 2018-10-24 Мерк Патент Гмбх Аминокислоты-липиды
CN104168888A (zh) * 2012-03-16 2014-11-26 默克专利股份有限公司 靶向氨基酸脂质
TWI586370B (zh) * 2012-03-16 2017-06-11 馬克專利公司 靶向胺基酸脂質
KR20140135832A (ko) * 2012-03-16 2014-11-26 메르크 파텐트 게엠베하 아미노산 지질
US9796666B2 (en) 2012-03-16 2017-10-24 Merck Patent Gmbh Aminoacid lipids
WO2013135360A1 (fr) * 2012-03-16 2013-09-19 Merck Patent Gmbh Lipides d'acides aminés
RU2654210C2 (ru) * 2012-03-16 2018-05-17 Мерк Патент Гмбх Нацеливающие аминокислотные липиды
US11510988B2 (en) 2012-03-16 2022-11-29 Merck Patent Gmbh Targeting aminoacid lipids
CN104168887A (zh) * 2012-03-16 2014-11-26 默克专利股份有限公司 氨基酸脂质
KR102081046B1 (ko) * 2012-03-16 2020-02-25 메르크 파텐트 게엠베하 아미노산 지질
US10111871B2 (en) 2013-08-27 2018-10-30 Northeastern University Nanoparticle drug delivery system and method of treating cancer and neurotrauma
US10342761B2 (en) 2014-07-16 2019-07-09 Novartis Ag Method of encapsulating a nucleic acid in a lipid nanoparticle host
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
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
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
CN116535381A (zh) * 2023-07-06 2023-08-04 北京悦康科创医药科技股份有限公司 具有五元环缩醛结构的阳离子脂质化合物、包含其的组合物及用途
CN116535381B (zh) * 2023-07-06 2023-10-17 北京悦康科创医药科技股份有限公司 具有五元环缩醛结构的阳离子脂质化合物、包含其的组合物及用途

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