111111
`
`1111111111111111111111111111111111111111111111111111111111111
`US007939505B2
`
`c12) United States Patent
`Quay et al.
`
`(10) Patent No.:
`(45) Date of Patent:
`
`US 7,939,505 B2
`May 10,2011
`
`(54) AMINO ACID LIPIDS AND USES THEREOF
`
`(75)
`
`Inventors: Steven C. Quay, Woodinville, WA (US);
`Michael Houston, Jr., Sannnamish, WA
`(US); Pierrot Harvie, Bothell, WA (US);
`Roger C. Adami, Snohomish, WA (US);
`Renata Fam, Kenmore, WA (US); Mary
`Gallagher Prieve, Lake Forest Park, WA
`(US); Kathy Lynn Fosnaugh, Boulder,
`CO (US); Shaguna Seth, Bothell, WA
`(US)
`
`(73) Assignee: Marina Biotech, Inc., Bothell, WA (US)
`
`( *) Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 162 days.
`
`(21) Appl. No.: 12/114,284
`
`(22) Filed:
`
`May2, 2008
`
`(65)
`
`Prior Publication Data
`
`US 2008/0317839 Al
`
`Dec. 25, 2008
`
`Related U.S. Application Data
`
`Provisional application No. 60/916,131, filed on May
`4, 2007, provisional application No. 60/947,282, filed
`on Jun. 29, 2007, provisional application No.
`60/953,667, filed on Aug. 2, 2007, provisional
`application No. 60/972,653, filed on Sep. 14, 2007,
`provisional application No. 60/972,590, filed on Sep.
`14, 2007, provisional application No. 61/022,571,
`filed on Jan. 22, 2008.
`
`Int. Cl.
`(2006.01)
`A61K 47114
`(2006.01)
`A61K 47118
`(2006.01)
`A61K 48100
`(2006.01)
`C07C 279114
`U.S. Cl. .................... 514/44 R; 514/44A; 514/785;
`514/788; 514/943; 554/53; 560/168; 564/159
`Field of Classification Search ........................ None
`See application file for complete search history.
`
`References Cited
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`
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`
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`
`(Continued)
`Primary Examiner- Jeffrey E Russel
`(74) Attorney, Agent, or Firm- Eckman Basu LLP
`ABSTRACT
`(57)
`This disclosure provides a range of amino acid lipid com(cid:173)
`pounds and compositions useful for drug delivery, therapeu(cid:173)
`tics, and the diagnosis and treatment of diseases and condi(cid:173)
`tions. The amino acid lipid compounds and compositions can
`be used for delivery of various agents such as nucleic acid
`therapeutics to cells, tissues, organs, and subjects.
`31 Claims, 5 Drawing Sheets
`
`Moderna Ex 1012-p. 1
`Moderna v Protiva
`
`

`

`US 7,939,505 B2
`Page 2
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`OTHER PUBLICATIONS
`Wang, K. et a!. "Synthesis and in vitro Behavior of Multivalent
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`* cited by examiner
`
`Moderna Ex 1012-p. 2
`Moderna v Protiva
`
`

`

`U.S. Patent
`
`May 10,2011
`
`Sheet 1 of 5
`
`US 7,939,505 B2
`
`FIG. 1
`
`Moderna Ex 1012-p. 3
`Moderna v Protiva
`
`

`

`U.S. Patent
`
`May 10,2011
`
`Sheet 2 of 5
`
`US 7,939,505 B2
`
`FIG. 2
`
`Moderna Ex 1012-p. 4
`Moderna v Protiva
`
`

`

`U.S. Patent
`
`May 10,2011
`
`Sheet 3 of 5
`
`US 7,939,505 B2
`
`FIG. 3
`
`A549 24 h PPIB mRNA Expression
`7,500 Cells/Well
`
`1.4
`
`1.2
`
`0.8
`
`0.6
`
`Q)
`:I
`
`iii >
`c:: c.
`
`CCI
`
`rf
`
`rn
`
`rf
`
`rf
`
`r1l
`
`1
`
`I 0.1
`
`I
`
`25 I 10
`
`I 1
`
`I 0.1
`
`Formulation 1
`
`Formulation 2
`
`siRNA (nM)
`
`RNAIMAX
`
`"C
`Q)
`.!::!
`iii 0.4
`E ...
`0
`z
`
`0.2
`
`0 n
`
`rfl
`25 I 10 I
`
`1
`
`0.1
`
`I
`
`......
`25
`
`...;;;;.
`
`10
`
`Moderna Ex 1012-p. 5
`Moderna v Protiva
`
`

`

`U.S. Patent
`
`May 10,2011
`
`Sheet 4 of 5
`
`US 7,939,505 B2
`
`FIG. 4
`
`9LIIacZ 72 h beta-Galactosidase Expression
`
`6,000 Cells/Well
`
`Formulation 1
`
`Formulation 2
`siRNA (nM)
`
`0.1
`
`RNAIMAX
`
`(!)
`
`0.05
`
`0.04
`
`0.03
`
`~ 0.06
`~
`Kl
`"C
`·;n
`~ Ctl
`Iii
`(!)
`cC..
`"C
`(!)
`-~
`Iii
`E ,_
`0 z
`
`0.02
`
`0.01
`
`Moderna Ex 1012-p. 6
`Moderna v Protiva
`
`

`

`U.S. Patent
`
`May 10,2011
`
`Sheet 5 of 5
`
`US 7,939,505 B2
`
`FIG. 5
`
`HepG2 24 h ApoB mRNA Expression
`15000 Cells/Well
`
`0.6
`
`G> :s 0.5
`ro >
`m 0.4
`0
`0..
`<(
`
`" 0.3
`G>
`-~
`(ij 0.2
`E ...
`0 z 0.1
`
`0
`
`25
`
`UNT
`
`Formulation 3
`
`Moderna Ex 1012-p. 7
`Moderna v Protiva
`
`

`

`US 7,939,505 B2
`
`1
`AMINO ACID LIPIDS AND USES THEREOF
`
`This application claims the benefit under 35 U.S.C.
`§119(e) of U.S. Provisional Application Nos. 61/022,571,
`filed Jan. 22, 2008, 60/972,590, filed Sep. 14, 2007, 60/972, 5
`653 filed Sep. 14, 2007, 60/953,667 filed Aug. 2, 2007,
`60/947,282 filed Jun. 29, 2007, and 60/916,131 filed May 4,
`2007, the contents of each of which is hereby incorporated by
`reference in its entirety.
`
`10
`
`BACKGROUND
`
`2
`acid lipids of this disclosure are molecules containing an
`amino acid residue and one or more lipophilic tails.
`In some aspects, this invention provides a range of amino
`acid lipid compounds as shown in Formula I:
`
`R3-(C=0)-Xaa-Z-R4
`
`Formula I
`
`wherein
`Xaa is any D- or L-amina acid residue having the formula
`-NRN---CR1R2-(C=0)-, or a peptide of n=2-20
`amino acid residues having the formula -{NRN(cid:173)
`CR1R2-(C=O)}n-, wherein
`R 1 is independently, for each occurrence, a non-hydro(cid:173)
`gen, substituted or unsubstituted side chain of an
`amino acid;
`R2 is independently, for each occurrence, hydrogen, or
`an organic group consisting of carbon, oxygen, nitro(cid:173)
`gen, sulfur, and hydrogen atoms, and having from 1 to
`20 carbon atoms, or C(1-5)alkyl, cycloalkyl,
`cycloalkylalkyl, C(3-5)alkenyl, C(3-5)alkynyl, C(1-
`5)alkanoyl, C(1-5)alkanoyloxy, C(1-5)alkoxy, C(1-5)
`alkoxy-C(1-5)alkyl,
`C(1-5)alkoxy-C(1-5)alkoxy,
`C(1-5)alkyl-amino-C(1-5)alkyl-,
`C(1-5)dialkyl(cid:173)
`amino-C(l-5)alkyl-, nitro-C(l-5)alkyl, cyano-C(1-5)
`alkyl, aryl-C(1-5)alkyl, 4-biphenyl-C(1-5)alkyl, car(cid:173)
`boxyl, or hydroxyl,
`RN is independently, for each occurrence, hydrogen, or
`an organic group consisting of carbon, oxygen, nitro(cid:173)
`gen, sulfur, and hydrogen atoms, and having from 1 to
`20 carbon atoms, or C(1-5)alkyl, cycloalkyl,
`cycloalkylalkyl, C(3-5)alkenyl, C(3-5)alkynyl, C(1-
`5)alkanoyl, C(1-5)alkanoyloxy, C(1-5)alkoxy, C(1-5)
`alkoxy-C(1-5)alkyl,
`C(1-5)alkoxy-C(1-5)alkoxy,
`C(1-5)alkyl-amino-C(1-5)alkyl-,
`C(1-5)dialkyl(cid:173)
`amino-C(l-5)alkyl-, nitro-C(l-5)alkyl, cyano-C(1-5)
`alkyl, aryl-C(1-5)alkyl, 4-biphenyl-C(1-5)alkyl, car(cid:173)
`boxyl, or hydroxyl,
`R3 is independently a lipophilic tail derived from a natu(cid:173)
`rally-occurring or synthetic lipid, phospholipid, gly(cid:173)
`colipid, triacylglycerol, glycerophospholipid, sphin(cid:173)
`golipid, ceramide, sphingomyelin, cerebroside, or
`ganglioside, wherein the tail may contain a steroid; or a
`substituted or unsubstituted C(3-22)alkyl, C(6-12)cy(cid:173)
`cloalkyl, C(6-12)cycloalkyl-C(3-22)alkyl, C(3-22)alk(cid:173)
`enyl, C(3-22)alkynyl, C(3-22)alkoxy, or C(6-12)
`alkoxy-C(3-22)alkyl;
`R4 is independently a lipophilic tail derived from a natu(cid:173)
`rally-occurring or synthetic lipid, phospholipid, gly(cid:173)
`colipid, triacylglycerol, glycerophospholipid, sphin(cid:173)
`golipid, ceramide, sphingomyelin, cerebroside, or
`ganglioside, wherein the tail may contain a steroid; or a
`substituted or unsubstituted C(3-22)alkyl, C(6-12)cy(cid:173)
`cloalkyl, C(6-12)cycloalkyl-C(3-22)alkyl, C(3-22)alk(cid:173)
`enyl, C(3-22)alkynyl, C(3-22)alkoxy, or C(6-12)
`alkoxy-C(3-22)alkyl;
`wherein either one of R3 and R4 is a lipophilic tail as
`defined above and the other is an amino acid terminal
`group, or both R3 and R4 are lipophilic tails; the amino
`acid terminal group being hydrogen, hydroxyl, amino,
`or an organic protective group;
`Z is NH, 0, S, -CH2S-, ---CH2 S(O)-, or an organic
`linker consisting of 1-40 atoms selected from hydrogen,
`carbon, oxygen, nitrogen, and sulfur atoms;
`and salts thereof.
`In some respects, this invention provides a range of amino
`65 acid lipid compounds as shown in Formula I:
`
`The delivery of a therapeutic compound to a subject can be
`impeded by limited ability of the compound to reach a target
`cell or tissue, or by restricted entry or trafficking of the com- 15
`pound within cells. Delivery of a therapeutic material is in
`general restricted by membranes of cells. These barriers and
`restrictions to delivery can result in the need to use much
`higher concentrations of a compound than is desirable to
`achieve a result, which brings the risk of toxic effects and side 20
`effects.
`One strategy for delivery is to improve transport of a com(cid:173)
`pound into cells using lipid or polymeric carrier molecules.
`These materials can take advantage of mechanisms that exist
`for selective entry into a cell, while still excluding exogenous 25
`molecules such as nucleic acids and proteins. For example, a
`cationic lipid may interact with a drug agent and provide
`contact with a cell membrane. Lipid molecules can also be
`organized into liposomes or particles as carriers for drug
`agents. Liposomal drug carriers can protect a drug molecule 30
`from degradation while improving its uptake by cells. Also,
`liposomal drug carriers can encapsulate or bind certain com(cid:173)
`pounds by electrostatic and other interactions, and may inter-
`act with negatively charged cell membranes to initiate trans(cid:173)
`port across a membrane.
`The understanding of regulatory RNA and the develop(cid:173)
`ment of RNA interference (RNAi), RNAi therapy, RNA
`drugs, antisense therapy, and gene therapy, among others, has
`increased the need for effective means of introducing active
`nucleic acid agents into cells. In general, nucleic acids are 40
`stable for only limited times in cells or plasma. However,
`nucleic acid-based agents can be stabilized in compositions
`and formulations which may then be dispersed for cellular
`delivery.
`This disclosure provides compounds, compositions, meth- 45
`ods and uses for improving systemic and local delivery of
`drugs and biologically active molecules.Among other things,
`this application provides novel compounds and compositions
`for making and using delivery structures and carriers which
`increase the efficiency of delivery ofbiologically active mol- 50
`ecules.
`
`35
`
`BRIEF SUMMARY
`
`This disclosure provides novel compounds, compositions 55
`and formulations for intracellular and in vivo delivery of drug
`agents for use, ultimately, as a therapeutic. The compounds
`and compositions of this disclosure are useful for delivery of
`drug agents to selected cells, tissues, organs or compartments
`in order to alter a disease state or a phenotype.
`In some aspects, this disclosure provides compounds, com(cid:173)
`positions and methods to deliver RNA structures to cells to
`produce the response of RNA interference, antisense effects,
`or the regulation of genomic expression.
`This invention provides a range of amino acid lipids which
`are lipophilic compounds for use in delivery and administra(cid:173)
`tion of drug agents and in drug delivery systems. The amino
`
`60
`
`R3 -(C=0)-Xaa-Z-R4
`
`Formula I
`
`Moderna Ex 1012-p. 8
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`

`US 7,939,505 B2
`
`3
`
`wherein
`Xaa is a D- or L-amina acid residue having the formula
`-NRN---CR1R2-(C=0)-, wherein
`R 1 is a substituted or unsubstituted basic side chain of an
`amino acid;
`R2 is hydrogen, or C(1-5)alkyl,
`RN is hydrogen, or C(1-5)alkyl,
`R3 is independently a substituted or unsubstituted C(6-22)
`alkyl or C(6-22)alkenyl;
`R4 is independently a substituted or unsubstituted C(6-22)
`alkyl or C(6-22)alkenyl;
`Z is NH, 0, or an organic linker consisting of 1-40 atoms
`selected from hydrogen,
`carbon, oxygen, nitrogen, and sulfur atoms;
`and salts thereof.
`In some embodiments, the amino acid lipid compound may
`contain Xaa selected from arginine, homoarginine, norargin(cid:173)
`ine, nor-norarginine, ornithine, lysine, homolysine, histidine,
`1-methylhistidine, pyridylalanine, asparagine, N-ethylaspar(cid:173)
`agine, glutamine, 4-aminophenylalanine, theN-methylated 20
`versions thereof, and side chain modified derivatives thereof.
`In some embodiments, the amino acid lipid compound may
`contain Xaa selected from cysteine and serine.
`In certain embodiments, R3 and R4 may be C(6-22)alkyl
`and may be the same or different. In some embodiments, R3 25
`and R4 may be C(6-22)alkenyl and may be the same or dif(cid:173)
`ferent.
`In certain embodiments, Xaa may be a peptide of 2-20
`amino acid residues.
`In some embodiments, Xaa may have a side chain contain- 30
`ing a functional group having a pKa from 5 to 7.5.
`In some aspects, the amino acid lipid compound may be a
`multi-mer of two or more of the amino acid lipid compounds
`which are crosslinked.
`In some embodiments, the amino acid lipid compound may 35
`be a conjugate having a peptide conjugated to the side chain
`of the amino acid residue.
`In certain embodiments, the amino acid lipid compound
`may be attached to an oligomeric or polymeric framework.
`In some embodiments, the amino acid lipid compound may 40
`be attached to a pharmaceutical drug compound.
`In some aspects, this invention provides a range of amino
`acid lipid compounds as shown in Formula I:
`
`4
`and one or more additional non-amino acid lipids or poly(cid:173)
`meric lipids. In some embodiments, the composition may
`contain cholesteryl hemisuccinate.
`In some aspects, this disclosure encompasses composi(cid:173)
`tions containing one or more amino acid lipid compounds and
`one or more nucleic acids which may form a complex with an
`amino acid lipid.
`In certain embodiments, this disclosure encompasses com(cid:173)
`positions containing one or more amino acid lipid compounds
`10 which form liposomes.
`In some aspects, this disclosure encompasses composi(cid:173)
`tions containing one or more amino acid lipid compounds
`which form an emulsion.
`In some embodiments, this disclosure encompasses com-
`15 positions containing one or more amino acid lipid compounds
`which form a micellar dispersion.
`In certain aspects, this disclosure encompasses composi(cid:173)
`tions containing one or more amino acid lipid compounds and
`one or more drug agents or biologically active agents.
`In some aspects, this disclosure encompasses methods for
`delivering a therapeutic nucleic acid to a cell by preparing a
`composition containing one or more amino acid lipid com(cid:173)
`pounds and treating a cell with the composition.
`In some embodiments, this disclosure encompasses meth(cid:173)
`ods for inhibiting expression of a gene in a cell comprising
`preparing a composition containing one or more amino acid
`lipid compounds and treating a cell with the composition.
`In certain aspects, this disclosure encompasses methods
`for inhibiting expression of a gene in a mammal comprising
`preparing a composition containing one or more amino acid
`lipid compounds and administering the composition to the
`mammal.
`In some embodiments, this disclosure encompasses meth(cid:173)
`ods for treating a disease in a human, the disease being
`selected from rheumatoid arthritis, liver disease, encephalitis,
`bone fracture, heart disease, viral disease including hepatitis
`and influenza, and cancer, comprising preparing a composi(cid:173)
`tion containing one or more amino acid lipid compounds and
`administering the composition to the human.
`In some aspects, this disclosure encompasses uses of a
`composition containing one or more amino acid lipid com(cid:173)
`pounds in the preparation of a medicament for treating a
`disease
`including
`rheumatoid arthritis,
`liver disease,
`encephalitis, bone fracture, heart disease, viral disease
`including hepatitis and influenza, and cancer.
`In some embodiments, this disclosure encompasses uses of
`a composition containing one or more amino acid lipid com(cid:173)
`pounds for treating a disease selected from rheumatoid arthri(cid:173)
`tis, liver disease, encephalitis, bone fracture, heart disease,
`50 viral disease including hepatitis and influenza, and cancer.
`This summary, taken along with the detailed description of
`the invention, as well as the figures, the appended examples
`and claims, as a whole, encompass the disclosure of the
`invention.
`
`45
`
`55
`
`BRIEF DESCRIPTION OF THE DRAWINGS
`
`FIG. 1: Schematic representation of a liposomal embodi(cid:173)
`ment of this invention in which amino acid lipids form a
`60 bilayer vesicle 10 along with other lipids. In this embodiment,
`the outer layer of the liposome is protected by polyethyleneg(cid:173)
`lycol chains 20 attached to a head group of one of the lipids.
`The outer layer of the liposome also presents a ligand 30 for
`specific targeting of a cell or tissue. The liposomal vesicle
`65 contains, in this embodiment, a cargo of active interfering
`RNA components including a condensed RNA nanoparticle
`40, a two-stranded RNA duplex peptide conjugate 50, a three-
`
`R3-(C=0)-Xaa-Z-R4
`
`Formula I
`
`wherein
`Xaa is a D- or L-amina acid residue having the formula
`-NRN---CR1R2-(C=0)-, wherein
`R 1 is a substituted or unsubstituted basic side chain of an
`amino acid;
`R2 is hydrogen, or C(1-5)alkyl,
`RN is hydrogen, or C(1-5)alkyl,
`R3 is independently a substituted or unsubstituted C(6-22)
`alkyl or C(6-22)alkenyl;
`R4 is hydrogen;
`Z is NH, 0, or an organic linker consisting of 1-40 atoms
`selected from hydrogen,
`carbon, oxygen, nitrogen, and sulfur atoms;
`and salts thereof.
`In some respects, this disclosure encompasses composi(cid:173)
`tions containing one or more amino acid lipid compounds and
`one or more therapeutic nucleic acids. The therapeutic
`nucleic acid may be a gene silencing agent, or an RNAi(cid:173)
`inducing agent, or a double-stranded RNA, or an mdRNA, or
`may contain a modified nucleoside.
`In some embodiments, this disclosure encompasses com(cid:173)
`positions containing one or more amino acid lipid compounds
`
`Moderna Ex 1012-p. 9
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`
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`

`US 7,939,505 B2
`
`5
`strandedmdRNA 60, a dicer substrate RNA 70, a dsRNA with
`a long overhang 80, and an siRNA with blunt ends 90, which
`are pooled in this embodiment.
`FIG. 2: Transmission electron micrograph obtained on a
`JEOL 1230 TEM of a liposomal embodiment of this invention
`showing spherical lipid bilayer vesicle particles formed with
`the amino acid lipid C12-norArg-C12. The length marker of
`the micrograph is 0.5 micrometer, and the sample was stained
`with 3% uranyl acetate. The lipid portion of the liposomal
`formulation was [C12-norArg(NH3 +cn---C12/DSPC/cho(cid:173)
`lesterol/DSPE-PEG-2000] in the amounts of [30%/20%/
`49%/1%] respectively, as a mole percent of total lipid. The
`liposomes contained the antiinfluenza-active dicer substrate
`dsRNA DX3030.
`FIG. 3: In FIG. 3 is shown an example of PPIB gene
`knockdown activity obtained from an in vitro assay inA549
`cells. The concentration response at 25, 10, 1, and 0.1 nM
`RNA of the normalized PPIB mRNA expression values for
`two amino acid lipid formulations of an interfering-RNA
`were compared to results for RNAIMAX. Formulation 1 was
`[C12-norArg(NH3 Cl)---C12/DOPE/CHOL (50/32/18)] and
`Formulation 2 was [C12-norArg(NH3 Cl)---C12/CHEMS/
`DLPE (50/32/18)]. The PPIB gene knockdown by an inter(cid:173)
`fering-RNA in an amino acid lipid composition of this dis(cid:173)
`closure can exceed that obtained with RNAIMAX.
`FIG. 4: In FIG. 4 is shown an example of LacZ gene
`knockdown activity obtained from an in vitro assay in 9
`L/LacZ cells. The concentration response at 25, 10, 1, and0.1
`nM RNA of the normalized beta-galactosidase expression
`values for two amino acid lipid formulations of an interfering(cid:173)
`RNA were compared to results for RNAIMAX. Formulation
`1 was [C12-norArg(NH3 Cl)---C12/DOPE/CHOL (50/32/
`18)] and Formulation 2 was [C12-norArg(NH3Cl)-C12/
`CHEMS/DLPE (50/32/18)]. The LacZ gene knockdown by
`an interfering-RNA in an amino acid lipid composition of this
`disclosure can exceed that obtained with RNAIMAX.
`FIG. 5: In FIG. 5 is shown an example of ApoB gene
`knockdown activity obtained from an in vitro assay in HepG 2
`cells. The concentration response at 25, 2.5, and 0.25 nM
`RNA of the normalized ApoB mRNA expression values for
`three amino acid lipid formulations of an interfering-RNA are
`shown. Formulation 1 was [non-amino acid cationic lipid/
`DSPC/chol./DMPE-PEG2k (40/10/48/2)]. Formulation 2
`and 3 were both
`[C18:1-norArg-C16/CHEMS/DLPE/
`DMPE-PEG2k (50/32/16/2)].
`
`DETAILED DESCRIPTION
`
`This disclosure relates generally to novel compounds,
`compositions and uses thereof for delivery of drug agents.
`The compounds and compositions of this disclosure are use-
`ful for delivery of therapeutic agents to selected cells, tissues,
`organs or subjects.
`This disclosure relates generally to the chemistry oflipids 55
`and uses of lipid-like structures and materials to effect drug
`delivery.
`This invention relates to novel drug delivery enhancing
`agents including lipids that are useful for delivering various
`molecules to cells. This invention provides a range of com- 60
`pounds, compositions, formulations, methods and uses of
`such agents directed ultimately toward drug delivery, thera(cid:173)
`peutics, and the diagnosis and treatment of diseases and con(cid:173)
`ditions, including those that respond to modulation of gene
`expression or activity in a subject. More specifically, this 65
`invention relates to compounds, liposomes, lamellar vesicles,
`emulsions, micelles, suspensions, particles, solutions and
`
`6
`other forms of delivery enhancing compositions and formu(cid:173)
`lations, as well as therapeutic methods and uses for these
`delivery materials.
`The compounds and compositions of this disclosure are
`useful for delivery of therapeutic, prophylactic, and diagnos(cid:173)
`tic agents such as nucleic acids, polynucleotides, peptides,
`proteins, and small molecule compounds and drugs.
`The compounds and compositions of this disclosure are
`useful for delivery of therapeutic agents in forms such as
`10 liposomes, lamellar vesicles, emulsions, micelles, suspen(cid:173)
`sions, particles, and solutions. These forms may include
`nanoparticles of various diameters.
`In some respects, the compounds and compositions of this
`15 disclosure are useful for delivery of a therapeutic agent in a
`liposome. In these embodiments the therapeutic agent may be
`referred to as the cargo. For example, FIG. 1 shows a sche(cid:173)
`matic representation of a liposomal embodiment of this
`invention in which amino acid lipids form a bilayer vesicle 10
`20 along with other lipids. In this embodiment, the outer layer of
`the liposome is protected by polyethyleneglycol chains 20
`attached to a head group of one of the lipids. The outer layer
`of the liposome also presents a ligand 30 for specific targeting
`of a cell or tissue. The liposomal vesicle contains, in this
`25 embodiment, a cargo of active interfering RNA components
`including a condensed RNA nanoparticle 40, a two-stranded
`RNA duplex peptide conjugate 50, a three-stranded mdRNA
`60, a dicer substrate RNA 70, a dsRNA with a long overhang
`80, and an siRNA with blunt ends 90, which are pooled in this
`30 embodiment. Other forms of therapeutic cargo may include
`microRNA or hairpin RNA forms.
`In some aspects, compounds and compositions of this dis(cid:173)
`closure may provide delivery of therapeutic agents in releas(cid:173)
`able forms or compositions. Releasable forms and composi-
`35 tions include molecules that bind and release an active agent,
`molecules that bind an active agent and discharge a moiety
`that assists in release of the agent, molecules that bind an
`active agent and are subsequently modulated in form within a
`biological compartment to assist in release of the agent, and
`40 compositions containing mole