Rl and R2 are independently selected from H, (Ci-C4)alkyl, heterocyclyl, and a polyamine, wherein said alkyl, heterocyclyl and polyamine are optionally substituted with one or more substituents selected from R', or Rl and R2 can be taken together with the nitrogen to which they are attached to form a monocyclic heterocyclyl with 4-7 members optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said monocyclic heterocyclyl is optionally substituted with one or more substituents selected from R';

R3 is absent or can be taken together with Rl to form a monocyclic heterocyclyl with 4-7 members optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said monocyclic heterocyclyl is optionally substituted with one or more substituents selected from R'; R' is independently selected from halogen, R", OR", SR", CN, C0₂R" and

CON(R")₂;

R" is selected from H and (Ci-C4)alkyl, wherein said alkyl is optionally substituted with one or more substituents selected from halogen and OH;

Li is a C4-C22 alkyl or C4-C22 alkenyl, said alkyl and alkenyl are optionally substituted with one or more substituents selected from R'; and

L2 is a C4-C22 alkyl or C4-C22 alkenyl, said alkyl and alkenyl are optionally substituted with one or more substituents selected from R';

or any pharmaceutically acceptable salt or stereoisomer thereof.

In a second embodiment, the invention features a compound having Formula A, wherein:

n is 1;

Y is a bond;

R3 is absent or can be taken together with Rl to form a monocyclic heterocyclyl with 4-7 members optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said monocyclic heterocyclyl is optionally substituted with one or more substituents selected from R';

R' is independently selected from halogen, R", OR", SR", CN, C0₂R" and

CON(R")₂;

or any pharmaceutically acceptable salt or stereoisomer thereof.

In a third embodiment, the invention features a compound having Formula A, wherein:

n is 1;

Y is a 0(C=0); Rl and R2 are independently selected from H, (Ci -Chalky 1, heterocyclyl, and a polyamine, wherein said alkyl, heterocyclyl and polyamine are optionally substituted with one or more substituents selected from R¹, or Rl and R2 can be taken together with the nitrogen to which they are attached to form a monocyclic heterocyclyl with 4-7 members optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said monocyclic heterocyclyl is optionally substituted with one or more substituents selected from R';

R' is independently selected from halogen, R", OR", SR", CN, C0₂R" and

CON(R")₂;

or any pharmaceutically acceptable salt or stereoisomer thereof.

In another embodiment of this invention, the cationic lipids are illustrated by the

Formula B:

wherein:

Rl and R2 are independently selected from H, (Ci-C4)alkyl, heterocyclyl, and a polyamine, wherein said alkyl, heterocyclyl and polyamine are optionally substituted with one or more substituents selected from R', or Rl and R can be taken together with the nitrogen to which they are attached to form a monocyclic heterocyclyl with 4-7 members optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said monocyclic heterocyclyl is optionally substituted with one or more substituents selected from R';

R' is independently selected from halogen, R", OR", SR", CN, C0₂R" and

CON(R")2; R" is selected from H and (Ci-C4)alkyl, wherein said alkyl is optionally substituted with one or more substituents selected from halogen and OH;

or any pharmaceutically acceptable salt or stereoisomer thereof.

Specific cationic lipids are:

N-[(l lZ)-icos-l 1-en-l-yl]- N' N'-dimethyl-N-nonylethane-l,2-diamine (Compound 1);

1 -methylpiperidin-4-yl decyl [(9Z)-octadec-9-en- 1 -yl] carbamate (Compound 2) ;

4-(dimethylamino)-N-[(9Z)-octadec-9-en- 1 -yl]-N-octylbutanamide (Compound 3);

l-methylpiperidin-4-yl decyl [(9Z)-octadec-9-en-l-yl] carbamate (Compound 4);

1 -methylpiperidin-4-yl decyl [(9Z,12Z)-octadeca-9,l 2-dien- 1-yl] carbamate (Compound 5);

N,N-dimethyl-N'-[(9Z, 12Z)-octadeca-9, 12-dien- 1 -yl]-N'-octylpropane- 1 ,3 -diamine

(Compound 6);

N,N-dimethyl-N'-nonyl-N'-[(9Z, 12Z)-octadeca-9, 12-dien- 1 -yljpropane- 1 ,3 -diamine

(Compound 7);

(9Z, 12Z)-N-[2-(pyrrolidin- 1 -yl)ethyl]-N-undecyloctadeca-9, 12-dien- 1 -amine (Compound 8); (1 lZ)-N-decyl-N-[2-(pyrrolidin-l-yl)ethyl]icos-l 1-en-l-amine (Compound 9);

N-decyl- 10- { 2- [(2-pentylcyclopropyl)methyl] cyclopropyl } -N- [2-(pyrrolidin- 1 -yl)ethyl] decan- 1 -amine (Compound 10);

N-(8-{2-[(2-pentylcyclopropyl)methyl]cyclopropyl}octyl)-N-[2-(pyrrolidin-l-yl)ethyl]decan- 1 -amine (Compound 11);

(1 lZ,14Z)-N-decyl-N-[2-(pyrrolidin-l-yl)ethyl]icosa-l 1,14-dien-l -amine (Compound 12); N-decyl-10-(2-octylcyclopropyl)-N-[2-(pyrrolidin-l-yl)ethyl]decan-l-amine (Compound 13); (9Z)-N-[(9Z)-octadec-9-en- 1 -yl]-N-[2-(pyrrolidin- 1 -yl)ethyl]octadec-9-en- 1 -amine

(Compound 14);

N,N-dimethyl-N',N'-di[(9Z)-octadec-9-en- 1 -yl]ethane- 1 ,2-diamine (Compound 15);

N,N-dimethyl-N',N'-di[(9Z, 12Z)-octadeca-9, 12-dien- 1 -yl]ethane- 1 ,2-diamine (Compound 16); N,N-dimethyl-N'-[(9Z)-octadec-9-en- 1 -yl]-N'-octylethane-l ,2-diamine (Compound 17);

(9Z)-N-octyl-N-[2-(pyrrolidin-l -yl)ethyl]octadec-9-en- 1 -amine (Compound 18);

N,N-dimethyl-N'-nonyl-N'-[(9Z, 12Z)-octadeca-9, 12-dien- 1 -yl] ethane- 1 ,2-diamine (Compound 19);

N-decyl-N',N'-dimethyl-N-[(9Z, 12Z)-octadeca-9, 12-dien- 1 -yl]ethane- 1 ,2-diamine (Compound 20);

N-decyl-N',N'-dimethyl-N-[(9Z)-octadec-9-en-l -yl]ethane-l ,2-diamine (Compound 21);

N,N-dimethyl-N'-nonyl-N'-[(9Z)-octadec-9-en-l-yl]ethane-l,2-diamine (Compound 22); and (9Z, 12Z)-N-decyl-N-[2-(pyrrolidin-l -yl)ethyl]octadeca-9, 12-dien- 1 -amine (Compound 23). or any pharmaceutically acceptable salt or stereoisomer thereof.

In another embodiment, the cationic lipids disclosed are useful in the preparation of lipid nanoparticles.

In another embodiment, the cationic lipids disclosed are useful components in a lipid nanoparticle for the delivery of oligonucleotides.

In another embodiment, the cationic lipids disclosed are useful components in a lipid nanoparticle for the delivery of siRNA and miRNA.

In another embodiment, the cationic lipids disclosed are useful components in a lipid nanoparticle for the delivery of siRNA.

The cationic lipids of the present invention may have asymmetric centers, chiral axes, and chiral planes (as described in: E.L. Eliel and S.H. Wilen, Stereochemistry of Carbon Compounds, John Wiley & Sons, New York, 1994, pages 1119-1190), and occur as racemates, racemic mixtures, and as individual diastereomers, with all possible isomers and mixtures thereof, including optical isomers, being included in the present invention. In addition, the cationic lipids disclosed herein may exist as tautomers and both tautomeric forms are intended to be encompassed by the scope of the invention, even though only one tautomeric structure is depicted.

It is understood that substituents and substitution patterns on the cationic lipids of the instant invention can be selected by one of ordinary skill in the art to provide cationic lipids that are chemically stable and that can be readily synthesized by techniques known in the art, as well as those methods set forth below, from readily available starting materials. If a substituent is itself substituted with more than one group, it is understood that these multiple groups may be on the same carbon or on different carbons, so long as a stable structure results.

It is understood that one or more Si atoms can be incorporated into the cationic lipids of the instant invention by one of ordinary skill in the art to provide cationic lipids that are chemically stable and that can be readily synthesized by techniques known in the art from readily available starting materials.

In the compounds of Formula A, the atoms may exhibit their natural isotopic abundances, or one or more of the atoms may be artificially enriched in a particular isotope having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature. The present invention is meant to include all suitable isotopic variations of the compounds of Formula A. For example, different isotopic forms of hydrogen (H) include protium (lH) and deuterium (¾). Protium is the predominant hydrogen isotope found in nature. Enriching for deuterium may afford certain therapeutic advantages, such as increasing in vivo half-life or reducing dosage requirements, or may provide a compound useful as a standard for characterization of biological samples.

Isotopically-enriched compounds within Formula A can be prepared without undue

experimentation by conventional techniques well known to those skilled in the art or by processes analogous to those described in the Scheme and Examples herein using appropriate isotopically-enriched reagents and/or intermediates.

As used herein, "alkyl" means a straight chain, cyclic or branched saturated aliphatic hydrocarbon having the specified number of carbon atoms.

As used herein, "alkenyl" means a straight chain, cyclic or branched unsaturated aliphatic hydrocarbon having the specified number of carbon atoms including but not limited to diene, triene and tetraene unsaturated aliphatic hydrocarbons.

Examples of a cyclic "alkyl" or "alkenyl are:

As used herein, "heterocyclyl" or "heterocycle" means a 4- to 10-membered aromatic or nonaromatic heterocycle containing from 1 to 4 heteroatoms selected from the group consisting of O, N and S, and includes bicyclic groups. "Heterocyclyl" therefore includes, the following: benzoimidazolyl, benzofuranyl, benzofurazanyl, benzopyrazolyl, benzotriazolyl, benzothiophenyl, benzoxazolyl, carbazolyl, carbolinyl, cinnolinyl, furanyl, imidazolyl, indolinyl, indolyl, indolazinyl, indazolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, naphthpyridinyl, oxadiazolyl, oxazolyl, oxazoline, isoxazoline, oxetanyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridopyridinyl, pyridazinyl, pyridyl, pyrimidyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl, tetrahydropyranyl, tetrazolyl, tetrazolopyridyl, thiadiazolyl, thiazolyl, thienyl, triazolyl, azetidinyl, 1 ,4-dioxanyl,

hexahydroazepinyl, piperazinyl, piperidinyl, pyrrolidinyi, morpholinyl, thiomorpholinyl, dihydrobenzoimidazolyl, dihydrobenzofuranyl, dihydrobenzothiophenyl, dihydrobenzoxazolyl, dihydrofuranyl, dihydroimidazolyl, dihydroindolyl, dihydroisooxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl, dihydrotetrazolyl,

dihydrothiadiazolyl, dihydrothiazolyl, dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl, methylenedioxybenzoyl, tetrahydrofuranyl, and tetrahydrothienyl, and N-oxides thereof all of which are optionally substituted with one to three substituents selected from R".

As used herein, "polyamine" means compounds having two or more amino groups. Examples include putrescine, cadaverine, spermidine, and spermine.

As used herein, "halogen" means Br, CI, F and I.

In an embodiment of Formula A, n is 1 or 2.

In an embodiment of Formula A, n is 2.

In an embodiment of Formula A, n is 1.

In an embodiment of Formula A, n is 0.

In an embodiment of Formula A, Y is a bond, 0(C=0), C=0, or NR'(C=0). In an embodiment of Formula A, Y is a bond.

In an embodiment of Formula A, Y is a bond or 0(C=0).

In an embodiment of Formula A, Y is 0(C=0).

In an embodiment of Formula A, Y is C=0.

In an embodiment of Formula A or B, Rl and R2 are independently selected from H and (Ci-C4)alkyl, wherein said alkyl is optionally substituted with one to three substituents selected from R', or Rl and R2 can be taken together with the nitrogen to which they are attached to form a monocyclic heterocyclyl with 4-7 members optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said monocyclic heterocyclyl is optionally substituted with one to three substituents selected from R'.

In an embodiment of Formula A or B, Rl and R2 are independently selected from H, methyl, ethyl and propyl, wherein said methyl, ethyl and propyl are optionally substituted with one to three substituents selected from R¹, or Rl and R2 can be taken together with the nitrogen to which they are attached to form a monocyclic heterocyclyl with 4-7 members optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said monocyclic heterocyclyl is optionally substituted with one to three substituents selected from R'.

In an embodiment of Formula A or B, Rl and R2 are independently selected from H, methyl, ethyl and propyl.

In an embodiment of Formula A or B, Rl and R2 are each methyl.

In an embodiment of Formula A, R3 is absent.

In an embodiment of Formula A, R3 is taken together with Rl to form a monocyclic heterocyclyl.

In an embodiment of Formula A or B, R' is R".

In an embodiment of Formula A or B, R" is independently selected from H, methyl, ethyl and propyl, wherein said methyl, ethyl and propyl are optionally substituted with one or more halogen and OH.

In an embodiment of Formula A or B, R" is independently selected from H, methyl, ethyl and propyl.

In an embodiment of Formula A or B, R" is H.

In an embodiment of Formula A or B, Li is selected from C4-C22 alkyl and C4-C22 alkenyl, which are optionally substituted with halogen and OH.

In an embodiment of Formula A or B, I is selected from C4-C22 alkyl and C4-C22 alkenyl.

In an embodiment of Formula A or B, Li is selected from C12-C22 alkyl and C12-C22 alkenyl. In an embodiment of Formula A or B, L\ is selected from C16-C20 alkyl and Cl6-C20 lkenyl.

In an embodiment of Formula A or B, Li is selected from Ci 8 alkyl and C\ 8 alkenyl.

In an embodiment of Formula A or B, L2 is selected from C4-C22 alkyl and C4-C22 alkenyl, which are optionally substituted with halogen and OH.

In an embodiment of Formula A or B, L2 is selected from C4-C22 alkyl and C4-C22 alkenyl.

In an embodiment of Formula A or B, L2 is selected from C6-C 18 alkyl and C6-C18 alkenyl.

In an embodiment of Formula A or B, L2 is selected from C6-C18 alkyl. In an embodiment of Formula A or B, L2 is selected from Cs-Cio alkyl. In an embodiment of Formula A or B, Li is

In an embodiment of Formula A or B, L\ is

In an embodiment of Formula A or B, I is

In an embodiment of Formula A or B, Li is

In an embodiment of Formula A or B, L2 is

K

In an embodiment of Formula A or B, L2 is

In an embodiment of Formula A or B, L2 is In an embodiment of Formula A or B, L2 is

In an embodiment of Formula A or B, Li and L2 are

In an embodiment of Formula A or B, L] and L2 are

In an embodiment of Formula A or B, "heterocyclyl" is pyrolidine, piperidine, morpholine, imidazole or piperazine.

In an embodiment of Formula A or B, "monocyclic heterocyclyl" is pyrolidine, piperidine, morpholine, imidazole or piperazine.

In an embodiment of Formula A or B, "polyamine" is putrescine, cadaverine, spermidine or spermine.

In an embodiment, "alkyl" is a straight chain saturated aliphatic hydrocarbon having the specified number of carbon atoms.

In an embodiment, "alkenyl" is a straight chain unsaturated aliphatic

hydrocarbon having the specified number of carbon atoms.

Included in the instant invention is the free form of cationic lipids of Formula A, as well as the pharmaceutically acceptable salts and stereoisomers thereof. Some of the isolated specific cationic lipids exemplified herein are the protonated salts of amine cationic lipids. The term "free form" refers to the amine cationic lipids in non-salt form. The encompassed pharmaceutically acceptable salts not only include the isolated salts exemplified for the specific cationic lipids described herein, but also all the typical pharmaceutically acceptable salts of the free form of cationic lipids of Formula A. The free form of the specific salt cationic lipids described may be isolated using techniques known in the art. For example, the free form may be regenerated by treating the salt with a suitable dilute aqueous base solution such as dilute aqueous NaOH, potassium carbonate, ammonia and sodium bicarbonate. The free forms may differ from their respective salt forms somewhat in certain physical properties, such as solubility in polar solvents, but the acid and base salts are otherwise pharmaceutically equivalent to their respective free forms for purposes of the invention.

The pharmaceutically acceptable salts of the instant cationic lipids can be synthesized from the cationic lipids of this invention which contain a basic or acidic moiety by conventional chemical methods. Generally, the salts of the basic cationic lipids are prepared either by ion exchange chromatography or by reacting the free base with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid in a suitable solvent or various combinations of solvents. Similarly, the salts of the acidic compounds are formed by reactions with the appropriate inorganic or organic base.

Thus, pharmaceutically acceptable salts of the cationic lipids of this invention include the conventional non-toxic salts of the cationic lipids of this invention as formed by reacting a basic instant cationic lipids with an inorganic or organic acid. For example, conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like, as well as salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic (TFA) and the like.

When the cationic lipids of the present invention are acidic, suitable

"pharmaceutically acceptable salts" refers to salts prepared form pharmaceutically acceptable non-toxic bases including inorganic bases and organic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic salts, manganous, potassium, sodium, zinc and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as arginine, betaine caffeine, choline, Ν,Ν¹- dibenzylethylenediamine, diethylamin, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine tripropylamine, tromethamine and the like.

The preparation of the pharmaceutically acceptable salts described above and other typical pharmaceutically acceptable salts is more fully described by Berg et al,

"Pharmaceutical Salts," J Pharm. Set, 1977:66:1-19.

It will also be noted that the cationic lipids of the present invention are potentially internal salts or zwitterions, since under physiological conditions a deprotonated acidic moiety in the compound, such as a carboxyl group, may be anionic, and this electronic charge might then be balanced off internally against the cationic charge of a protonated or alkylated basic moiety, such as a quaternary nitrogen atom.

EXAMPLES

Examples provided are intended to assist in a further understanding of the invention. Particular materials employed, species and conditions are intended to be further illustrative of the invention and not limitative of the reasonable scope thereof. The reagents utilized in synthesizing the cationic lipids are either commercially available or are readily prepared by one of ordinary skill in the art.

Compounds derived from General Scheme 1 can be generally obtained from employing the following strategy. A diamine i is combined with a carboxylic acid ii and appropriate coupling agent to furnish an amino-secondary-amide intermediate iii. The carbonyl of this intermediate is subsequently reduced using LAH to furnish a new diamine iv. This compound is once again reacted with a carboxylic acid and coupling agent to provide an amino-tertiary-amide v. Reduction of the amide carbonyl yields the final diamine cationic lipid vi.

GENERAL SCHEME 1

Compounds derived from General Scheme 2 can be generally obtained from employing the following strategy. The amine intermediate viii undergoes standard amide coupling with a carboxylic acid vii followed by LAH reduction to yield a key secondary amine intermediate x Finally, the amine is reacted with an acyl electrophile such as an activated carboxylic acid or chloroformate to furnish an amide or carbamates xi, respectively.

GENERAL SCHEME 2

VII VIM

N'-idlZMcos-ll-en-l-yll-N. N-dimethylethane-1, 2-diamine (Intermediate A)

To a 500 mL DCM solution of oleic acid (20.0 g, 70.8 mmol) was added CDI (11.5 g, 70.8 mmol), and the solution stirred at RT for 1 hour, after which N,N-dimethylethane-l, 2-diamine (6.24 g, 70.8 mmol) was added. After 1 hour the reaction was washed with 300 mL water, then with brine. The organic was dried over Na₂S0₄, filtrated and the solvent evaporated in vacuo to yield a colorless oil. The oil was taken up in 500 mL THF and cooled to 0^°C in an ice bath. Lithium aluminium hydride 2.0 M solution in THF (53.2 mL, 106 mmol) was added slowly, dropwise. The ice bath was removed, and the reaction was allowed to gradually warm to RT over 2 hours. The reaction was then diluted with 100 mL more THF and was quenched by adding small portions of sodium sulfate dodecahydrate until fizzing stopped. Solids were filtered off using a Buchner funnel, and the filtrate was concentrated in vacuo to yield

Intermediate A (20.5 g). LCMS (m + H): 339.5

N-[2-dimethvIamiLne)ethvI1-N-[(llZ)-icos-ll-eii-l-yllnonamide (Intermediate B)

To a 30 mL THF solution of N'-[(l lZ)-icos-l l-en-l-yl]-N, N-dimethylethane-1, 2-diamine (2.50 g, 7.38 mmol) and nonanoic acid (1.29 g, 8.12 mmol) was added diisopropylethylamine (1.15 g, 8.86 mmol) and HATU (3.23 g, 8.49 mmol). The reaction stirred at RT for 1 hour, after which it was washed with 30 mL of saturated NH₄C1 solution, NaHC0₃ solution, brine, respectively. The organic was dried over Na₂S0₄, filtrated and purified by silica gel chromatography (0% ethyl acetate / hexane→ 100% ethyl acetate) to give Intermediate B

(3.02 g). LCMS (m + H): 479.7

N-[(llZ)-icos-ll-en-l-yll- N' N'-dimethyl-N-nonylethane-l^-diamine (Compound 1)

To 30 ml THF solution of N-[2-dimethylamine)ethyl]-N-[(l lZ)-icos-l l-en-l-yl]nonamide

o

(3.02 g, 6.31 mmol) at 0 C was added lithium aluminum hydride 2.0 M solution in THF (6.31 mL, 12.6 mmol) slowly, dropwise. The reaction was stirred and allowed to gradually warm to RT over 2 hours. The reaction was diluted in THF, and sodium sulfate dodecahydrate was added in small portions until fizzing stopped. Solids were removed by filtration through a Buchner funnel. The filtrate was concentrated to give N-[(l lZ)-icos-l 1-en-l-yl]- N' N- dimethyl-N-nonylethane-l,2-diamine (1) (1.89 g). HRMS (m + H): 463.4979. Ή NMR δ (ppm)( CDC1₃): 5.39 - 5.31 (2H, m), 2.56 - 2.53 (2H, m), 2.43 - 2.37 (6H, m) 2.24 - 2.22 (6H, m), 2.07 - 2.01 (4H, m), 1.81 (2H, m), 1.46 - 1.41 (4H, m), 1.40 - 1.20 (30H, m), 0.90 - 0.84 (6H, m).

(9Z)-N-decyoctadec-9-enamide (Intermediate Q

To a 500 mL DCM solution of) oleic acid (20.0 g, 70.8 mmol) was added CDI (11.48 g, 70.8 mmol), and the solution stirred for 1 hour at RT. Decylamine (11.14 g, 70.8 mmol) was added, and the reaction stirred another 16 hours at RT. The solution was washed with water and washed with brine. The organic layer was dried over Na₂S0₄, and purified by silica gel chromatography (0% ethyl acetate / hexane→ 100% ethyl acetate) to give Intermediate C

(13.0 g). LCMS (m + l): 422.6

(9Z)-N-decyloctadec— en- 1 -amine (Intermediate D)

A 250 mL THF solution of (9Z)-N-decyoctadec-9-amide (28.1 g, 66.6 mmol) was cooled to

0 C in an ice bath. Litium aluminum hydride 2.0 M solution in THF (66.6 mL, 133 mmol) was added slowly, dropwise. The ice bath was removed, and the reaction was allowed to come to RT over 2 hours. The reaction was further diluted with 100 mL more of THF, and sodium sulfate dodecahydrate was added in small portions until fizzing stopped. Solids were removed using a Buchner funnel, and the filtrate was concentrated in vacuo to yield Intermediate D

(26.7 g). LCMS (m + 1): 408.6 l-methylpiperidin-4-yl decvU(9Z)-octadec-9-en-l-yll carbamate (Compound 2)

To a solution of 4-hydroxy-l-methylpiperidine (0.52 mL, 4.41 mmol) in 25 mL DCM was added triethylamine (1.03 mL, 7.36 mmol) at 0°C. 4-Nitrophenyl chloroformate (0.89 g, 4.41 mmol) in 3 mL of DCM was added dropwise. The cold bath was removed, and the reaction was warmed to RT over 4 hours. (9Z)-N-decyloctadec— en- 1 -amine was added, and the reaction was heated to 30°C for 19 hours. The reaction was cooled to RT, washed with water, washed with brine, dried over Na₂S0₄, and purified by silica gel chromatography (0% acetone / hexane→ 15% acetone/hexane with 2% TEA) to yield compound (2) (0.90 g). HRMS (m + H): 549.5345. Ή NMR δ (ppm)( CDC1₃): 5.36 - 5.32 (2H, m), 4.73 - 4.71 (1H, m), 3.18 (4H, br s), 2.59 (2H, m), 2.29 (5H, m), 2.02 - 1.99 (4H, m), 1.94 - 1.90 (2H, m), 1.77 - 1.69 (2H), 1.55 - 1.50 (4H), 1.39 - 1.20 (38H, m), 0.88 - 0.86 (6H, m).

Compounds 3-23 were prepared in a manner analogous to that described for compounds 1 and 2 above:

Me

Me

19 N^-dimethyl-N'-nonyl-jV'- [(9Z, 12Z)-octadeca- 463.4986 463.4979 9, 12-dien- 1 -yljethane- 1 ,2-diamine

20 N-decyl-iV_yV-dimethyl-N-[(9Z, 12Z)-octadeca- 477.5142 477.5138

9, 12-dien- 1 -yll ethane- 1 ,2-diamine

21 N-decyl-iV '-dimethyl-N-[(9Z)-octadec-9-en- 479.5299 479.5294

1 -yl]ethane- 1 ,2-diamine

22 N,N-dimethyl-iV-nonyl-iV-[(9Z)-octadec-9-en- 465.5142 465.5138

1 -yl] ethane- 1 ,2-diamine

23 (9Z, 12Z)-N-decyl-N- [2-(pyrrolidin- 1 - 503.5299 503.5291 yl)ethyl]octadeca-9, 12-dien- 1 -amine

Compound 24 is DLinDMA as described in J. Controlled Release, 2005, 107, 276-287, US 2006/0083780 Al, and US 2006/0008910 Al.

Compound 25 is DLinKC2DMA as described in Nature Biotechnology, 2010, 28, 172-176, WO 2010/042877 Al, WO 2010/048536 A2, WO 2010/088537 A2, and WO 2009/127060 Al.

(25)

Compounds 26-30 were prepared in a manner analogous to that described for compounds 1 and 2 above:

27

28

29 o__{cc r}

30

o HRMS data available. Compound confirmed by HPLC.

o HRMS data available. 1H NMR d (ppm)(400 MHz, CDC13): 5.36-5.34 (m, 2H), 2.61- 1 (m, 8H), 2.44-2.40 (m, 4H), 2.04-1.97 (m, 4H), 1.79-1.76 (m, 4H), 1.46-1.41 (m, 4H), 6 (m, 36H), 0.88 (t, J = 6.68 Hz, 6H).

o HRMS data available. LC/MS (M+H) LNP COMPOSITIONS

The following lipid nanoparticle compositions (LNPs) of the instant invention are useful for the delivery of oligonucleotides, specifically siRNA and miRNA:

Cationic Lipid / Cholesterol / PEG-DMG 56.6/38/5.4;

Cationic Lipid / Cholesterol / PEG-DMG 60/38/2;

Cationic Lipid/ Cholesterol / PEG-DMG 67.3/29/3.7;

Cationic Lipid / Cholesterol / PEG-DMG 49.3/47/3.7;

Cationic Lipid / Cholesterol / PEG-DMG 50.3/44.3/5.4;

Cationic Lipid / Cholesterol / PEG-C-DMA / DSPC 40/48/2/10;

Cationic Lipid / Cholesterol / PEG-DMG / DSPC 40/48/2/10; and

Cationic Lipid / Cholesterol / PEG-DMG / DSPC 58/30/2/10.

LNP process description:

The Lipid Nano-Particles (LNP) are prepared by an impinging jet process. The particles are formed by mixing lipids dissolved in alcohol with siRNA dissolved in a citrate buffer. The mixing ratio of lipids to siRNA are targeted at 45-55% lipid and 65-45% siRNA. The lipid solution contains a novel cationic lipid of the instant invention, a helper lipid

(cholesterol) , PEG (e.g. PEG-C-DMA, PEG-DMG) lipid, and DSPC at a concentration of 5-15 mg/mL with a target of 9-12 mg/mL in an alcohol (for example ethanol). The ratio of the lipids has a mole percent range of 25-98 for the cationic lipid with a target of 35-65, the helper lipid has a mole percent range from 0-75 with a target of 30-50, the PEG lipid has a mole percent range from 1-15 with a target of 1-6, and the DSPC has a mole precent range of 0-15 with a target of 0-12. The siRNA solution contains one or more siRNA sequences at a concentration range from 0.3 to 1 .0 mg/mL with a target of 0.3 -0.9 mg/mL in a sodium citrate buffered salt solution with pH in the range of 3.5-5. The two liquids are heated to a temperature in the range of 15-40°C, targeting 30-40°C, and then mixed in an impinging jet mixer instantly forming the LNP. The teelD has a range from 0.25 to 1.0 mm and a total flow rate from 10 - 600 mL/min. The combination of flow rate and tubing ID has effect of controlling the particle size of the LNPs between 30 and 200 nm. The solution is then mixed with a buffered solution at a higher pH with a mixing ratio in the range of 1 : 1 to 1 :3 vol:vol but targeting 1 :2 vol:vol. This buffered solution is at a temperature in the range of 15-40°C, targeting 30-40°C. The mixed LNPs are held from 30 minutes to 2 hrs prior to an anion exchange filtration step. The temperature during incubating is in the range of 15-40°C, targeting 30-40°C. After incubating the solution is filtered through a 0.8 um filter containing an anion exchange separation step. This process uses tubing IDs ranging from 1 mm ID to 5 mm ID and a flow rate from 10 to 2000 mL/min. The LNPs are concentrated and diafiltered via an ultrafiltration process where the alcohol is removed and the citrate buffer is exchanged for the final buffer solution such as phosphate buffered saline. The ultrafiltration process uses a tangential flow filtration format (TFF). This process uses a membrane nominal molecular weight cutoff range from 30 -500 KD. The membrane format can be hollow fiber or flat sheet cassette. The TFF processes with the proper molecular weight cutoff retains the LNP in the retentate and the filtrate or permeate contains the alcohol; citrate buffer; final buffer wastes. The TFF process is a multiple step process with an initial concentration to a siRNA concentration of 1 -3 mg/mL. Following concentration, the LNPs solution is diafiltered against the final buffer for 10 -20 volumes to remove the alcohol and perform buffer exchange. The material is then concentrated an additional 1-3 fold. The final steps of the LNP process are to sterile filter the concentrated LNP solution and vial the product.

Analytical Procedure:

1) siRNA concentration

The siRNA duplex concentrations are determined by Strong Anion-Exchange High-Performance Liquid Chromatography (SAX-HPLC) using Waters 2695 Alliance system (Water Corporation, Milford MA) with a 2996 PDA detector. The LNPs, otherwise referred to as RNAi Delivery Vehicles (RDVs), are treated with 0.5% Triton X-l 00 to free total siRNA and analyzed by SAX separation using a Dionex BioLC DNAPac PA 200 (4 χ 250 mm) column with UV detection at 254 nm. Mobile phase is composed of A: 25 mM NaC10₄, 10 mM Tris, 20% EtOH, pH 7.0 and B: 250 mM NaC10₄, 10 mM Tris, 20% EtOH, pH 7.0 with liner gradient from 0-15 min and flow rate of 1 ml/min. The siRNA amount is determined by comparing to the siRNA standard curve.

2) Encapsulation rate

Fluorescence reagent SYBR Gold is employed for RNA quantitation to monitor the encapsulation rate of RDVs. RDVs with or without Triton X-100 are used to determine the free siRNA and total siRNA amount. The assay is performed using a SpectraMax M5e microplate spectrophotometer from Molecular Devices (Sunnyvale, CA). Samples are excited at 485 nm and fluorescence emission was measured at 530 nm. The siRNA amount is determined by comparing to the siRNA standard curve.

Encapsulation rate = (1- free siRNA/total siRNA) l00%

3) Particle size and polydispersity

RDVs containing 1 μg siRNA are diluted to a final volume of 3 ml with 1^χ

PBS. The particle size and polydispersity of the samples is measured by a dynamic light scattering method using ZetaPALS instrument (Brookhaven Instruments Corporation, Holtsville, NY). The scattered intensity is measured with He-Ne laser at 25°C with a scattering angle of 90°.

4) Zeta Potential analysis

RDVs containing 1 μg siRNA are diluted to a final volume of 2 ml with 1 mM Tris buffer (pH 7.4). Electrophoretic mobility of samples is determined using ZetaPALS instrument (Brookhaven Instruments Corporation, Holtsville, NY) with electrode and He-Ne laser as a light source. The Smoluchowski limit is assumed in the calculation of zeta potentials. 5 Lipid analysis

Individual lipid concentrations are determined by Reverse Phase High- Performance Liquid Chromatography (RP-HPLC) using Waters 2695 Alliance system (Water Corporation, Milford MA) with a Corona charged aerosol detector (CAD) (ESA Biosciences, Inc, Chelmsford, MA). Individual lipids in RDVs are analyzed using an Agilent Zorbax SB- C18 (50 x 4.6 mm, 1.8 μιη particle size) column with CAD at 60 °C. The mobile phase is composed of A: 0.1% TFA in H₂0 and B: 0.1% TFA in IP A. The gradient changes from 60% mobile phase A and 40% mobile phase B from time 0 to 40% mobile phase A and 60% mobile phase B at 1.00 min; 40% mobile phase A and 60% mobile phase B from 1.00 to 5.00 min; 40% mobile phase A and 60% mobile phase B from 5.00 min to 25% mobile phase A and 75% mobile phase B at 10.00 min; 25%» mobile phase A and 75% mobile phase B from 10.00 min to 5% mobile phase A and 95% mobile phase B at 15.00 min; and 5% mobile phase A and 95% mobile phase B from 15.00 to 60% mobile phase A and 40% mobile phase B at 20.00 min with flow rate of 1 ml/min. The individual lipid concentration is determined by comparing to the standard curve with all the lipid components in the RDVs with a quadratic curve fit. The molar percentage of each lipid is calculated based on its molecular weight.

Utilizing the above described LNP process, specific LNPs with the following ratios were identified:

Nominal composition:

Cationic Lipid / Cholesterol / PEG-DMG 60/38/2

Cationic Lipid / Cholesterol / PEG-DMG / DSPC 58/30/2/10

Luc siRNA

5 '-iB- A C/AAGGC 6Ά t/G A AG AG A C/ATT-iB 3' (SEQ.ID.NO.: 1) - UAUUCCGAUACUUCUC A}J-5' (SEQ.ID.NO.:2)

AUGC - Ribose

iB - Inverted deoxy abasic

UC- 2' Fluoro

AGT - 2^* Deoxy

AGU - 2* OCH₃

Nominal composition

Cationic Lipid /Cholesterol/PEG-DMG 60/38/2

Cationic Lipid / Cholesterol / PEG-DMG / DSPC 40/48/2/10

Cationic Lipid / Cholesterol / PEG-DMG / DSPC 58/30/2/10

ApoB siRNA

54B-CUUU^C^4UUCCU04 4UTsT-iB-3' (SEQ ID NO.:3) 3'-UsUG^4^UE/GUU^4GG,4CUsUsUsA-5' (SEQ ID NO.:4)

AUGC - Ribose

iB - Inverted deoxy abasic

UC- 2' Fluoro

AGT - 2' Deoxy

AGU - 2' OCH₃

UsA - phophorothioate linkage

Oligonucleotide synthesis is well known in the art. (See US patent applications: US 2006/0083780, US 2006/0240554, US 2008/0020058, US 2009/0263407 and US

2009/0285881 and PCT patent applications: WO 2009/086558, WO2009/ 127060,

WO2009/132131, WO2010/042877, WO2010/054384, WO2010/054401, WO2010/054405 and WO2010/054406). The siRNAs disclosed and utilized in the Examples were synthesized via standard solid phase procedures.

EXAMPLE 1

Mouse In Vivo Evaluation of Efficacy

LNPs utilizing Compound 1-23, in the nominal compositions described immediately above, were evaluated for in vivo efficacy. The siRNA targets the mRNA transcript for the firefly (Photinus pyralis) luciferase gene (Accession # Ml 5077). The primary sequence and chemical modification pattern of the luciferase siRNA is displayed above. The in vivo luciferase model employs a transgenic mouse in which the firefly luciferase coding sequence is present in all cells. ROSA26- LoxP-Stop-LoxP-Luc (LSL-Luc) transgenic mice licensed from the Dana Farber Cancer Institute are induced to express the Luciferase gene by first removing the LSL sequence with a recombinant Ad-Cre virus (Vector Biolabs). Due to the organo-tropic nature of the virus, expression is limited to the liver when delivered via tail vein injection. Luciferase expression levels in liver are quantitated by measuring light output, using an IVIS imager (Xenogen) following administration of the luciferin substrate (Caliper Life Sciences). Pre-dose luminescence levels are measured prior to administration of the RDVs. Luciferin in PBS (15mg/mL) is intraperitoneally (IP) injected in a volume of 150 μί. After a four minute incubation period mice are anesthetized with isoflurane and placed in the IVIS imager. The RDVs (containing siRNA) in PBS vehicle were tail vein injected n a volume of 0.2 mL. Final dose levels ranged from 0.1 to 0.5 mg/kg siRNA. PBS vehicle alone was dosed as a control. Mice were imaged 48 hours post dose using the method described above.

Changes in luciferin light output directly correlate with luciferase mRNA levels and represent an indirect measure of luciferase siRNA activity. In vivo efficacy results are expressed as % inhibition of luminescence relative to pre-dose luminescence levels. Systemic administration of the luciferase siRNA RDVs decreased luciferase expression in a dose dependant manner. Similar efficacy was observed in mice dosed with Compound 1 containing RDVs than with the RDV containing the DLinDMA (Compound 24) cationic lipid (Figure 1). OCD is known and described in WO2010/021865.

Rat In Vivo Evaluation of Efficacy and Toxicity

LNPs utilizing compounds in the nominal compositions described above, were evaluated for in vivo efficacy and increases in alanine amino transferase and aspartate amino transferase in Sprague-Dawley (Crl:CD(SD) female rats (Charles River Labs). The siRNA targets the mRNA transcript for the ApoB gene (Accession # NM 019287). The primary sequence and chemical modification pattern of the ApoB siRNA is displayed above. The RDVs (containing siRNA) in PBS vehicle were tail vein injected in a volume of 1 to 1.5 mL. Infusion rate is approximately 3 ml/min. Five rats were used in each dosing group. After LNP administration, rats are placed in cages with normal diet and water present. Six hours post dose, food is removed from the cages. Animal necropsy is performed 24 hours after LNP dosing. Rats are anesthetized under isoflurane for 5 minutes, then maintained under anesthesia by placing them in nose cones continuing the delivery of isoflurane until ex-sanguination is completed. Blood is collected from the vena cava using a 23 gauge butterfly venipuncture set and aliquoted to serum separator vacutainers for serum chemistry analysis. Punches of the excised caudate liver lobe are taken and placed in RNALater (Ambion) for mRNA analysis. Preserved liver tissue was homogenized and total RNA isolated using a Qiagen bead mill and the Qiagen miRNA-Easy RNA isolation kit following the manufacturer's instructions. Liver ApoB mRNA levels were determined by quantitative RT-PCR. Message was amplified from purified RNA utilizing a rat ApoB commercial probe set (Applied Biosystems Cat #

RN01499054_ml). The PCR reaction was performed on an ABI 7500 instrument with a 96- well Fast Block. The ApoB mRNA level is normalized to the housekeeping PPIB (NM 011149) mRNA. PPIB mRNA levels were determined by RT-PCR using a commercial probe set (Applied Biosytems Cat. No. Mm00478295_ml). Results are expressed as a ratio of ApoB mRNA/ PPIB mRNA. All mRNA data is expressed relative to the PBS control dose. Serum ALT and AST analysis were performed on the Siemens Advia 1800 Clinical Chemistry Analyzer utilizing the Siemens alanine aminotransferase (Cat# 03039631) and aspartate aminotransferase (Cat# 03039631) reagents. Similar efficacy and improved tolerability were observed in rats dosed with Compound 1 containing RDV than with the RDV containing the cationic lipid DLinKC2DMA (Compound 25, Figure 2).

Determination of Cationic Lipid Levels in Rat Liver

Liver tissue was weighed into 20-ml vials and homogenized in 9 v/w of water using a GenoGrinder 2000 (OPS Diagnostics, 1600 strokes/min, 5min). A 50 aliquot of each tissue homogenate was mixed with 300 of extraction/protein precipitating solvent (50/50 acetonitrile/methanol containing 500 nM internal standard) and the plate was centrifuged to sediment precipitated protein. A volume of 200 μΐ, of each supernatant was then transferred to separate wells of a 96-well plate and 10 μΐ samples were directly analyzed by LC/MS-MS.

Standards were prepared by spiking known amounts of a methanol stock solution of Compound 1 or OCD (OCD is known and described in WO2010/021865) into untreated rat liver homogenate (9 vol water/weight liver). Aliquots (50 μΐ,) each standard/liver homogenate was mixed with 300 μΐ, of extraction/protein precipitating solvent (50/50 acetonitrile/methanol containing 500 nM internal standard) and the plate was centrifuged to sediment precipitated protein. A volume of 200 μί of each supernatant was transferred to separate wells of a 96-well plate and 10 μΐ of each standard was directly analyzed by LC/MS- MS.

Absolute quantification versus standards prepared and extracted from rat liver homogenate was performed using an Aria LX-2 HPLC system (Thermo Scientific) coupled to an API 4000 triple quadrupole mass spectrometer (Applied Biosystems). For each run, a total of 10 μΐ, sample was injected onto a BDS Hypersil C8 HPLC column (Thermo, 50 x 2mm, 3 μιη) at ambient temperature (Figure 3).

Mobile Phase A: 95% H20/5% methanol/10 mM ammonium

formate/O.P/oformic acid Mobile Phase B: 40% methanol/60% n-propanol/10 mM ammonium formate/0. l%formic acid The flow rate was 0.5 mL/min and gradient elution profile was as follows: hold at 80% A for 0.25 min, linear ramp to 100% B over 1.6 min, hold at 100% B for 2.5 min, then return and hold at 80% A for 1.75 min. Total run time was 5.8 min. API 4000 source parameters were CAD: 4, CUR: 15, GS1 : 65, GS2: 35, IS: 4000, TEM: 550, CXP: 15, DP: 60, EP: 10.

Claims

WHAT IS CLAIMED IS:

1. A cationic lipid of Formula A:

A wherein: n is 1 or 2;

Y is a bond, 0(C=0), C=0, or NR'(CO);

Rl and R2 are independently selected from H, (Ci-C4)alkyl, heterocyclyl, and a polyamine, wherein said alkyl, heterocyclyl and polyamine are optionally substituted with one or more substituents selected from R', or Rl and R2 can be taken together with the nitrogen to which they are attached to form a monocyclic heterocyclyl with 4-7 members optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said monocyclic heterocyclyl is optionally substituted with one or more substituents selected from R ;

R' is independently selected from halogen, R", OR", SR", CN, C0₂R" and CON(R")2;

L2 is a C4-C22 alkyl or C4-C22 alkenyl, said alkyl and alkenyl are optionally substituted with one or more substituents selected from R'; or any pharmaceutically acceptable salt or stereoisomer thereof. 2. A cationic lipid of Formula A according to Claim 1, wherein: n is 1;

Y is a bond;

Rl and R.

2 are independently selected from H, (Ci-C4)alkyl, heterocyclyl, and a polyamine, wherein said alkyl, heterocyclyl and polyamine are optionally substituted with one or more substituents selected from R', or Rl and R2 can be taken together with the nitrogen to which they are attached to form a monocyclic heterocyclyl with 4-7 members optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said monocyclic heterocyclyl is optionally substituted with one or more substituents selected from R;

L2 is a C4-C22 alkyl or C4-C22 alkenyl, said alkyl and alkenyl are optionally substituted with one or more substituents selected from R'; or any pharmaceutically acceptable salt or stereoisomer thereof.

3. A cationic lipid of Formula A according to Claim 1 , wherein: n is 1 ;

Y is a 0(C=0);

I I and R.2 are independently selected from H, (Ci-C4)alkyl, heterocyclyl, and a polyamine, wherein said alkyl, heterocyclyl and polyamine are optionally substituted with one or more substituents selected from R', or Rl and R2 can be taken together with the nitrogen to which they are attached to form a monocyclic heterocyclyl with 4-7 members optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, 0 and S, said monocyclic heterocyclyl is optionally substituted with one or more substituents selected from R';

Li is a C4-C22 alkyl or C4-C22 alkenyl, said alkyl and alkenyl are optionally substituted with one or more substituents selected from R¹; and L2 is a C4-C22 alkyl or C4-C22 alkenyl, said alkyl and alkenyl are optionally substituted with one or more substituents selected from R'; or any pharmaceutically acceptable salt or stereoisomer thereof.

4. A cationic lipid of Formula B:

wherein:

R1 and R2 are independently selected from H, (Ci-C4)alkyl, heterocyclyl, and a polyamine, wherein said alkyl, heterocyclyl and polyamine are optionally substituted with one or more substituents selected from R', or Rl and R2 can be taken together with the nitrogen to which they are attached to form a monocyclic heterocyclyl with 4-7 members optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said monocyclic heterocyclyl is optionally substituted with one or more substituents selected from R';

Li is a C4-C22 alkyl or C4-C22 alkenyl, said alkyl and alkenyl are optionally substituted with one or more substituents selected from R'; and L2 is a C4-C22 alkyl or C4-C22 alkenyl, said alkyl and alkenyl are optionally substituted with one or more substituents selected from R'; or any pharmaceutically acceptable salt or stereoisomer thereof.

5. A cationic lipid which is:

N-[(l lZ)-icos-l 1-en-l-yl]- N' N'-dimethyl-N-nonylethane-l,2-diamine;

1 -methylpiperidin-4-yl decyl[(9Z)-octadec-9-en- 1 -yl]carbamate;

4-(dimethylamino)-N-[(9Z)-octadec-9-en- 1 -yl]-N-octylbutanamide;

l-methylpiperidin-4-yl decyl[(9Z)-octadec-9-en-l-yl]carbamate;

1 -methylpiperidin-4-yl decyl [(9Z, 12Z)-octadeca-9, 12-dien- 1 -yljcarbamate;

Ν,Ν-dimethyl-N'- [(9Z, 12Z)-octadeca-9, 12-dien- 1 -yl]-N'-octylpropane- 1 ,3 -diamine;

N,N-dimethyl-N'-nonyl-N'-[(9Z, 12Z)-octadeca-9, 12-dien- 1 -yl]propane- 1 ,3 -diamine;

(9Z, 12Z)-N-[2-(pyrrolidin- 1 -yl)ethyl]-N-undecyloctadeca-9, 12-dien- 1 -amine; (1 lZ)-N-decyl-N-[2-(pyrrolidin-l-yl)ethyl]icos-l 1-en-l-amine;

N-decyl- 10- { 2- [(2-pentylcyclopropyl)methyl] cyclopropyl } -N- [2-(pyrrolidin- 1 -yl)ethyl] decan- 1 -amine;

N-(8- { 2- [(2-pentylcyclopropyl)methyl] cyclopropyl } octyl)-N- [2-(pyrrolidin- 1 -yl)ethyl] decan- 1 -amine;

( 11 Z, 14Z)-N-decyl-N-[2-(pyrrolidin- 1 -yI)ethyl]icosa- 11,14-dien- 1 -amine;

N-decyl- 10-(2-octylcyclopropyl)-N-[2-(pyrrolidin- 1 -yl)ethyl]decan- 1 -amine;

(9Z)-N-[(9Z)-octadec-9-en-l -yl]-N-[2-(pyrrolidin- 1 -yl)ethyl]octadec-9-en- 1 -amine;

N,N-dimethyl-N',N'-di[(9Z)-octadec-9-en- 1 -yl]ethane- 1 ,2-diamine;

N,N-dimethyl-N',N'-di[(9Z, 12Z)-octadeca-9, 12-dien- 1 -yl]ethane- 1 ,2-diamine;

Ν,Ν-dimethyl-N'- [(9Z)-octadec-9-en- 1 -yl]-N'-octylethane- 1 ,2-diamine;

(9Z)-N-octyl-N-[2-(pyrrolidin-l-yl)ethyl]octadec-9-en-l-amine;

N,N-dimethyl-N'-nonyl-N'-[(9Z, 12Z)-octadeca-9, 12-dien- 1 -yl]ethane- 1 ,2-diamine;

N-decyl-N',N'-dimethyl-N- [(9Z, 12Z)-octadeca-9, 12-dien- 1 -yl]ethane- 1 ,2-diamine;

N-decyl-N',N'-dimethyl-N-[(9Z)-octadec-9-en-l -yl]ethane-l ,2-diamine;

N,N-dimethyl-N'-nonyl-N'-[(9Z)-octadec-9-en-l-yl]ethane-l,2-diamine;

(9Z, 12Z)-N-decyl-N-[2-(pyrrolidin- 1 -yl)ethyl]octadeca-9, 12-dien- 1 -amine;

(9Z, 12Z)-N-nonyl-N-[2-(pyrrolidin-l -yl)ethyl]octadeca-9, 12-dien- 1 -amine;

(9Z)-N-decyl-N-[2-(pyrrolidin- 1 -yl)ethyl]octadec-9-en- 1 -amine;

N-[8-(2-octylcyclopropyl)octyl]-N-[2-(pyrrolidin-l-yl)ethyl]decan-l-amine;

(9Z)-N-decyl-N- [2-(pyrrolidin- 1 -yl)ethyl] exadec-9-en- 1 -amine; and

N-decyl-N^l,N'-dimethyl-N-[8-(2-octylcyclopropyl)octyl]ethane-l,2-diamine; or any pharmaceutically acceptable salt or stereoisomer thereof.