Synthetic lipid materials for brain delivery

Abstract

Disclosed are (i) a compound of Formula I or a pharmaceutically acceptable salt thereof; and (ii) a lipidoid nanoparticle comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, and their use as a drug delivery vehicle across the blood brain barrier.

Description

[0001] Related Applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 019,530, filed May 4, 2020; the contents of which are incorporated by reference in their entirety.

[0003] Government Support

[0004] This invention was made with government support under Grant Numbers TR002636 and EB027170 awarded by the National Institutes of Health. The government has certain rights in the invention. BACKGROUND

[0005] Treatment of central nervous system (CNS) diseases such as neurodegenerative disorders, brain tumors, brain infections, and stroke is severely hampered by the blood brain barrier (BBB), which impedes the transport of most small molecule drugs and macromolecules (e.g., peptide, gene, and protein drugs) into the brain. Extensive efforts have been made to improve brain delivery efficiency so far, including direct CNS administration, disruption of the BBB, and carrier vehicle-mediated delivery. However, direct administration to the CNS is invasive, which can cause infection and tissue damage, and is also limited by the diffusion distance and rapid efflux of drugs out of the CNS within hours. Disruption of the BBB using techniques such as osmotic, biochemical, and ultrasound-mediated disruption can effectively introduce drugs into the brain, however, these transient openings of the BBB also allow plasma proteins to infiltrate the brain, leading to neurotoxicity, vasculopathy, and chronic neuropathology of the brain. Therefore, there is still a desire for methods to safely and effectively deliver BBB-impermeable cargos, particularly for gene and nucleic acid therapies, into the CNS.

[0006] Carrier vehicle-mediated drug delivery to the brain is considered a promising and versatile brain delivery system. Over the decades, various carrier vehicles such as viral vectors, exosomes, molecular Trojan horses, and various nanoparticle formulations have been developed to enhance brain delivery. Viral vectors are effective for delivering genes to the brain, but have limitations such as production cost and safety concerns. Exosomes have been used to deliver small molecules, proteins, and nucleic acids to the brain due to their non-immunogenicity; however, there are still many challenges in the isolation method, cargo loading procedure, in vivo toxicity, and pharmacokinetics. Molecular Trojan horse approach relies on receptor-specific monoclonal antibodies or peptides to ferry gene-fused cargos to the brain, which is promising in delivering biologics across the BBB. However, the production process needs to be tailored for each different biological cargo, and stability, safety, and immunogenicity are challenges for clinical development. Various nanoparticles such as liposomes, cationic polymers, inorganic nanoparticles, and nanocapsules have shown promise in delivering various cargos to the CNS, but always require complex modifications to ensure that the produced particles are BBB-permeable.

[0007] Neurotransmitters are endogenous chemicals that enable neural transmission. Notably, some neurotransmitters have been shown to cross the BBB. For example, dimethyltryptamine and other tryptamine derivatives have been shown to cross the BBB through active transport across the endothelial cell plasma membrane. SUMMARY

[0008] Disclosed herein is a simple and effective method for delivering cargos to the brain using neurotransmitter-derived synthetic lipids. The method is very robust and can be used to successfully deliver different classes of cargos (small molecules, nucleic acids, and proteins, etc.) all using the same simple nanoparticle design.

[0009] In one aspect, disclosed is a compound of the formula:

[0010] Y-W-R 脂质 (I),

[0011] or a pharmaceutically acceptable salt thereof, wherein

[0012] Y is a moiety derived from a neurotransmitter;

[0013] W is -NR 20 -, -O-, or -S-;

[0014] R 脂质 is independently substituted or unsubstituted C 1-20 alkyl, substituted or unsubstituted C 1-20 alkenyl, substituted or unsubstituted C 1-20 alkynyl, substituted or unsubstituted C 1-20 heteroalkyl, substituted or unsubstituted C1-20 heteroalkenyl or substituted or unsubstituted C 1-20 heteroalkynyl; and

[0015] R 20 R 脂质 , H, C 1-6 alkyl, C 1-6 alkenyl or C 1-6 alkynyl.

[0016] In certain aspects, disclosed are lipidoid nanoparticles comprising a compound disclosed herein.

[0017] In certain aspects, disclosed are pharmaceutical compositions comprising a lipidoid nanoparticle disclosed herein; and a pharmaceutically acceptable carrier or excipient. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 A is a schematic of NT-lipid-doped LNP formulated for brain delivery of cargo.

[0019] Figure 1 B is a synthetic route of neurotransmitter for lipidoid synthesis, lipid nomenclature, and chemical structure schematic.

[0020] Figure 1 C is a representative in vitro fluorescence image of a brain dissected 1 hour post DiR-labeled NT-LNP. DiR was doped into NT-LNP at a 10% weight ratio. Mice were perfused with normal saline prior to dissection. -1 DiR-labeled NT-LNP. DiR was doped into NT-LNP at a 10% weight ratio. Mice were perfused with normal saline prior to dissection.

[0021] Figure 2 A is the chemical structure of PBA-Q76-O16B, NT1-O12B, and a schematic of NT1-lipid-doped AmB formulation.

[0022] Figure 2 B is a photograph of AmB formulations doped with different amounts of PBA-Q76-O16B (using weight ratio) in NT1-O12B. Pure NT1-O12B / AmB encapsulates appear as opaque suspensions, while the appearance of the encapsulates changes from a translucent solution to a uniform transparent yellow solution as the doping ratio of PBA-Q76-O16B lipidoid increases.

[0023] Figure 2 C is a graph depicting the hydrodynamic diameter and polydispersity index of different NT-LNP / AmB formulations determined by DLS measurements.

[0024] Figure 2 D is a representative in vivo fluorescence image of a brain dissected 1 hour post DiR-labeled NT-LNP. DiR was doped into NT-LNP at a 10% weight ratio. Mice were perfused with normal saline prior to dissection. -1Representative fluorescence images of dissected mouse brains 1 hour post NT1-O12B / PBA-Q76-O16B DiR-LNP.

[0025] Figure 2 E is a plot depicting AmB concentration in brain tissue 24 hours post 5 mg / kg AmB in various NT1-O12B / PBA-Q76-O16B LNP LNP formulations via intravenous injection, measured with HPLC (n = 4 per group). Mice were perfused with saline prior to dissection. One-way ANOVA, Sidak’s post-hoc multiple analysis, *p < 0.05, **p < 0.001 or ***p < 0.0001. Graphical data represented as box plots with points overlaid, where error bars represent maximum and minimum values, and box lines represent median.

[0026] Figure 3 A shows the chemical structure of 306-O12B-3, NT1-O14B, and a schematic of the doped NT-lipid Tau-ASO formulation for brain delivery.

[0027] Figure 3 B is a plot depicting the GFP silencing efficiency of HEK-GFP cells treated with or without ASO / NT-LNP complexes. NT1-O14B LNPs alone do not show silencing efficacy, while doping NT1-lipid in 306-O12B-3 LNPs leads to successful in vitro gene silencing. *p < 0.01 compared to all other samples in the same group.

[0028] Figure 3 C is a plot depicting Tau-ASO formulated with NT1-O14B doped with different ratios of 306-O12B-3, saline, or scrambled Tau-ASO-LNPs via tail vein intravenous injection into C57BL / 6J mice (n = 6 per group) and analyzed for total tau mRNA levels in the brain. Graphical data represented as box plots with points overlaid, where error bars represent maximum and minimum values, and box lines represent median, *p < 0.05 or **p < 0.001.

[0029] Figure 3 D is a plot depicting total tau protein levels in the NT1-O14B / 306-O12B-3 = 3:7 group compared to saline or scrambled Tau-ASO levels, **p < 0.001. One-way ANOVA, Sidak’s post-hoc multiple analysis.

[0030] Figure 4 A is a schematic of mixed LNP formulations using NT1-O14B and PBA-Q76-O16B for delivery of GFP-Cre protein into the brain.

[0031] Figure 4 B is a fluorescent image of brain sections from Ai14 mice treated with (-27)GFP-Cre in different LNP formulations. Ai14 mice were intravenously injected with (-27)GFP-Cre complexed with LNP at NT1-O14B / PBA-Q76-O16B = 3:7, 10:0, or 0:10. After 3 weeks, the NT1-O14B / PBA-Q76-O16B = 3:7 group showed expression of tdTomato, indicating Cre-mediated recombination in the cerebral cortex, hippocampus, and cerebellum. Scale bar: 100 pm.

[0032] Figure 5 is a table of TEM images and hydrodynamic size, polydispersity index, zeta potential of NT1-LNP.

[0033] Figure 6 is a graph summarizing the concentration of AmB in brain tissue 24 hours after intravenous injection of 5 mg / kg AmB in various NT1 derivatives measured by HPLC. Mice were perfused with saline before dissection. -1 Graph of the relative fluorescence intensity of dissected brain tissue 1 hour after DiR-labeled NT-LNP. DiR was doped into NT-LNP at 10% weight ratio. Mice were perfused with saline before dissection. One-way ANOVA, Sidak’s post-hoc multiple analysis, *p < 0.05 or **p < 0.01.

[0034] Figure 7 is a representative in vitro fluorescence image of dissected brain 1 hour after DiR-labeled LNP or NT-LNP doped with NT1-O12B at a ratio of 3:7, w / w, and the chemical structures of 76-O16B, EC16-80, and 113-O16B. DiR was doped into NT-LNP at 10% weight ratio. Mice were perfused with saline before dissection. -1 Graph of the relative fluorescence intensity of dissected brain tissue 1 hour after DiR-labeled NT-LNP. DiR was doped into NT-LNP at 10% weight ratio. Mice were perfused with saline before dissection. One-way ANOVA, Sidak’s post-hoc multiple analysis, *p < 0.05 or **p < 0.01.

[0035] Figure 8 depicts the chemical structures of NT-lipid and dimethyltryptamine, and the concentration of AmB in brain tissue 24 hours after intravenous injection of 5 mg / kg AmB in various NT1 derivatives measured by HPLC. Mice were perfused with saline before dissection. -1 Graph of the relative fluorescence intensity of dissected brain tissue 1 hour after DiR-labeled NT-LNP. DiR was doped into NT-LNP at 10% weight ratio. Mice were perfused with saline before dissection. One-way ANOVA, Sidak’s post-hoc multiple analysis, *p < 0.05 or **p < 0.01.

[0036] Figure 9 is a graph depicting the concentration of AmB in brain tissue 24 hours after intravenous injection of 5 mg / kg AmB in various NT1 derivatives measured by HPLC. Mice were perfused with saline before dissection.

[0037] Figure 10A is a photograph of AmB formulations in NT1 class lipids with different tail lengths (018B, 016B, 014B, 012B). All four NT1 / AmB encapsulates appear as opaque suspensions.

[0038] Figure 10 B is a graph depicting the hydrodynamic diameter and polydispersity index of NT-LNPs determined by DLS measurements.

[0039] Figure 11 is a TEM image of NT1-O12B / PBA-Q76O16B-3 / 7-AmB complex, and a table summarizing the hydrodynamic size, polydispersity index, zeta potential, and DLC of AmB / NT-LNP complexes.

[0040] Figure 12 is a graph summarizing the relative fluorescence intensity of brain tissue dissected 1 hour after a single intravenous injection of 1 mg kg -1 is a graph summarizing the relative fluorescence intensity of brain tissue dissected 1 hour after a single intravenous injection of 1 mg kg

[0041] Figure 13 is a calibration curve of AmB concentration ranging from 0.005-0.5 ug / mL (low concentration) or 0.007-3.0 ug / mL (high concentration) dissolved in methanol at 415 nm wavelength by HPLC.

[0042] Figure 14 is a mAU-time graph of AmB concentration by HPLC 24 hours after intravenous treatment with NT1-O12B / PBA-Q76O16-LNP (ratio: 3 / 7)-AmB complex at a single dose of 5 mg AmB / kg.

[0043] Figure 15 is a graph depicting AmB concentration in other organs measured by HPLC 24 hours after intravenous injection of 5 mg / kg AmB.

[0044] Figure 16 is a TEM image of blank and ASO-loaded NT1-O14B / 306-O12B-3 (ratio: 3 / 7) nanoparticles, and a table of hydrodynamic size, polydispersity index, zeta potential.

[0045] Figure 17 is a TEM image of blank and (-27)GFP-Cre-loaded NT1-O14B / PBA-Q76O16B (ratio: 3 / 7) nanoparticles, and a table of hydrodynamic size, polydispersity index, zeta potential.

[0046] Figure 18A is a scheme depicting the synthesis of 1E tail.

[0047] Figure 18B is a scheme depicting the synthesis of PBA-Q76O16B and PBA-Q80O16B.

[0048] Figure 18C is a scheme depicting the synthesis of NT1-Neu.

[0049] Figures 19A-19N is a fluorescent image of Ai14 mouse brain sections. Mice were injected with Cre mRNA complexed with Dlin-MC3 / NT1-O14B LNP. LNP formulation is described in section 1.

[0050] Figures 20A-20B is a fluorescent image of Ai14 mouse brain sections. Mice were injected with Cre mRNA complexed with PBA-Q76O16B / NT1-O14B LNP. LNP formulation is described in section 1.

[0051] Figures 21A-21B is a fluorescent image of Ai14 mouse brain sections. Mice were injected with Cre mRNA complexed with Dlin-MC3 / NT1-O14B LNP. LNP formulation is described in section 1. DETAILED DESCRIPTION

[0052] In one aspect, disclosed is a compound of Formula I:

[0053] Y-W-R 脂质 (I),

[0054] or a pharmaceutically acceptable salt thereof, wherein:

[0055] Y is a moiety derived from a neurotransmitter;

[0056] W is -NR 20 -, -O-, or -S-;

[0057] R 脂质 is, independently, substituted or unsubstituted C 1-20 alkyl, substituted or unsubstituted C 1-20 alkenyl, substituted or unsubstituted C 1-20 alkynyl, substituted or unsubstituted C 1-20 heteroalkyl, substituted or unsubstituted C 1-20 heteroalkenyl, or substituted or unsubstituted C 1-20 heteroalkynyl; and R 20 is R 脂质 , H, C 1-6 alkyl, C 1-6 alkenyl, or C1-6 Alkyne group.

[0058] In some implementations, Y is selected from:

[0059]

[0060] In some preferred embodiments, Y is

[0061]

[0062] In some implementations, W is -NR 20 - or -S-. In some implementations, W is -NR. 20 - In some implementations, W stands for -S-.

[0063] In some implementations, W is -NR 20 - and R 20 For R 脂质 .

[0064] In some implementations, W is -NR 20 - and R 20 For R 脂质 And Y is

[0065]

[0066] In some implementations, R 脂质 The structure is as follows:

[0067]

[0068] in:

[0069] R 1 and R 2 Each instance is independently -H, -OH, -NHR 30 Or -SH;

[0070] R 3 and R 4 Both are -H; or R 3 and R 4 Together they form an oxo (=O) group;

[0071] Z represents -CH2-, -O-, or -NR. 30 -or-S-;

[0072] X and Y are independently -CH2- and -NR. 30 -, -O-, -S- or -Se-;

[0073] m is an integer selected from 1 to 3;

[0074] n is an integer selected from 1-14;

[0075] p is 0 or 1 ;

[0076] q is an integer selected from 1-10;

[0077] t is 0 or 1 ; and

[0078] R 30 is -H, C 1-6 1-6 alkyl, C 1-6 2-6 alkenyl, or C 1-6 2-6 alkynyl.

[0079] In certain embodiments, each instance of R 1 and R 2 is independently -H and -OH. In certain embodiments, R 1 and R 2 are -H. In certain embodiments, R 1 is -H; and R 2 is -OH.

[0080] In certain embodiments, R 3 and R 4 are -H. In certain embodiments, R 3 and R 4 together form an oxo (=0) group.

[0081] In certain embodiments, Z is -CH2-, -0-, or -NR 30 - In certain embodiments, Z is -CH2-. In certain embodiments, Z is -0-. In certain embodiments, Z is -NR 30 -.

[0082] In certain embodiments, R 1 and R 2 are -H, R 3 and R 4 together form an oxo (=0) group, and Z is O.

[0083] In certain embodiments, R 1 is -H, R 2 is -OH, R 3 and R 4 are -H, and Z is -CH2-.

[0084] In certain embodiments, X and Y are independently -CH2- or -O-. In certain embodiments, X and Y are independently -CH2- or -O-, wherein X and Y are not the same. In certain embodiments, X and Y are independently -CH2- or -S-. In certain embodiments, both X and Y are -CH2-. In certain embodiments, both X and Y are -S-.

[0085] In certain embodiments, m is 1 or 2. In certain embodiments, m is 1. In certain embodiments, m is 2.

[0086] In certain embodiments, n is an integer selected from 4-12. In certain embodiments, n is an integer selected from 6-10.

[0087] In certain embodiments, p is 0. In certain embodiments, p is 1.

[0088] In certain embodiments, q is an integer selected from 2-8. In certain embodiments, q is an integer selected from 4-8.

[0089] In certain embodiments, t is 0. In certain embodiments, t is 1.

[0090] In certain embodiments, the compound is selected from:

[0091]

[0092]

[0093]

[0094] or a pharmaceutically acceptable salt thereof.

[0095] In certain aspects, disclosed are lipidoid nanoparticles comprising a compound disclosed herein.

[0096] In certain embodiments, the nanoparticles disclosed herein further comprise a protein.

[0097] In certain embodiments, the protein is GFP-Cre.

[0098] In certain embodiments, the nanoparticles disclosed herein further comprise a nucleic acid.

[0099] In certain embodiments, the nucleic acid is a Tau-ASO.

[0100] In certain embodiments, the nanoparticles disclosed herein further comprise a small molecule.

[0101] In certain embodiments, the small molecule is an antifungal agent or a chemotherapeutic agent.

[0102] In certain embodiments, the small molecule is selected from bortezomib, imatinib, gefitinib, erlotinib, afatinib, osimertinib, dacomitinib, daunorubicin hydrochloride, cytarabine, fluorouracil, irinotecan hydrochloride, vincristine sulfate, methotrexate, paclitaxel, vincristine sulfate, epirubicin, docetaxel, cyclophosphamide, carboplatin, lenalidomide, ibrutinib, abiraterone acetate, enzalutamide, pemetrexed, palbociclib, nilotinib, everolimus, rebeccamycin, epirubicin, pirarubicin, idarubicin, valrubicin, amrubicin, bleomycin, patulin, dactinomycin, plicamycin, streptozotecin, pentostatin, mitosanes mitomycin C, enediyne calicheamicin, glycoside doxorubicin, macrolide epotihilone, ixabepilone, pentostatin, salinosporamide A, vinblastine, vincristine, etoposide, teniposide, vinorelbine, docetaxel, camptothecin, belinostat, cryptophycin, theopederin, annamide, trichostatin, aplidine, and ecteinascidin 743 (ET743).

[0103] In certain embodiments, the small molecule is amphotericin B or doxorubicin.

[0104] In certain embodiments, the lipidoid nanoparticle has a particle size of about 25 nm to about 1000 nm. In certain embodiments, the lipidoid nanoparticle has a particle size of about 50 nm to about 500 nm.

[0105] In certain aspects, disclosed are pharmaceutical compositions comprising the lipidoid nanoparticle disclosed herein; and a pharmaceutically acceptable carrier or excipient.

[0106] Definitions

[0107] Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings that are commonly understood by those of ordinary skill in the art. Generally, the terms and techniques described herein that are associated with chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics, and protein and nucleic acid chemistry are those that are well known and commonly used in the art.

[0108] The methods and techniques of the present disclosure are typically performed according to conventional methods well known in the art and as described in various general and more specific references which are cited throughout this specification unless otherwise indicated. See, e.g., "Principles of Neural Science", McGraw-Hill Medical, New York, N.Y. (2000); Motulsky, "Intuitive Biostatistics", Oxford University Press, Inc. (1995); Lodish et al., "Molecular Cell Biology, 4th Ed.", W.H. Freeman & Co., New York (2000); Griffiths et al., "Introduction to Genetic Analysis, 7th Ed.", W.H. Freeman & Co., N.Y. (1999); and Gilbert et al., "Developmental Biology, 6th Ed.", Sinauer Associates, Inc., Sunderland, MA (2000).

[0109] Unless otherwise defined, the chemical terms used herein are employed according to their conventional meanings within the art, as exemplified by "The McGraw-Hill Dictionary of Chemical Terms", Parker S., Ed., McGraw-Hill, San Francisco, C.A. (1985).

[0110] As used herein, the term "optional" or "optionally" means that the subsequently described event or circumstance can or can not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted alkyl" means that the alkyl can be substituted and that the alkyl is unsubstituted.

[0111] It will be appreciated that substituents and substitution patterns on the compounds of the application can be selected by one of ordinary skill in the art to result in chemically stable compounds that can be readily synthesized from readily available starting materials by techniques known in the art and those methods set forth below. If a substituent is itself substituted with more than one group, it will be understood that the multiple groups can be on the same carbon or on different carbons, as long as a stable structure results.

[0112] As used herein, the term "optionally substituted" means replacing 1-6 hydrogen groups in a given structure with a specified substituent, including but not limited to: hydroxyl, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl, cycloalkyl, heterocyclic, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH2-O-alkyl, -OP(O)(O-alkyl)2, or -CH2-OP(O)(O-alkyl)2. Preferably, "optionally substituted" means replacing 1-4 hydrogen groups in a given structure with the substituents mentioned above. More preferably, 1-3 hydrogen groups are replaced with the substituents mentioned above. It should be understood that the substituents may be further substituted.

[0113] Articles such as “a,” “an,” and “the” can mean one or more than one, unless the contrary is indicated or otherwise apparent from the context. A claim or description containing “or” among one or more members of the group is considered satisfied if one, more than one, or all of the group members are present in, used in, or otherwise associated with the given product or method, or are used in, or are otherwise associated with the given product or method. The invention includes embodiments in which exactly one member of the group is present in, used in, or otherwise associated with the given product or method. The invention includes embodiments in which more than one member or all of the group members are present in, used in, or otherwise associated with the given product or method.

[0114] As used herein, the term "alkyl" refers to a saturated aliphatic group, including but not limited to C1-C1 groups. 10 Straight-chain alkyl groups or C1-C 10 Branched alkyl group. Preferably, the "alkyl" group refers to a C1-C6 straight-chain alkyl group or a C1-C6 branched alkyl group. Most preferably, the "alkyl" group refers to a C1-C4 straight-chain alkyl group or a C1-C4 branched alkyl group. Examples of "alkyl" include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, 1-pentyl, 2-pentyl, 3-pentyl, neopentyl, 1-hexyl, 2-hexyl, 3-hexyl, 1-heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1-octyl, 2-octyl, 3-octyl, or 4-octyl. The "alkyl" group may optionally be substituted.

[0115] The term "acyl" is recognized in the art and refers to a group represented by the general formula hydrocarbon C(O)-, preferably alkyl C(O)-.

[0116] The term "amido" is art-recognized and refers to an amino group substituted by an acyl group, and can be represented, for example, by the formula hydrocarbylC(O)NH-.

[0117] The term "acyloxy" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0118] The term "alkoxy" refers to an alkyl group with an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propyloxy, t-butyloxy, and the like.

[0119] The term "alkoxyalkyl" refers to an alkyl group substituted by an alkoxy group, and can be represented by the general formula alkyl-O-alkyl.

[0120] The term "alkyl" refers to saturated aliphatic groups including straight-chain alkyl, branched-chain alkyl, cyclic (cycloalkyl) groups, alkyl-substituted cyclic alkyl groups, and cyclic alkyl-substituted alkyl groups. In preferred embodiments, straight-chain or branched-chain alkyl groups have 30 or fewer carbon atoms in their backbone (e.g., C 1-30 , for straight-chain and C 3-30 , for branched-chain), and more preferably 20 or fewer.

[0121] Further, as used throughout this specification, the term "alkyl" is intended to encompass both unsubstituted and substituted alkyl groups, the latter of which refers to alkyl moieties having substituents replacing one or more hydrogens on the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, and the like.

[0122] The term "C x-y " or "C x -C y " when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is intended to include groups containing from x to y carbons in the chain. Co alkyl indicates a hydrogen if the group is at a terminal position, or a bond if the group is internal. For example, C 1-6 alkyl groups contain 1-6 carbon atoms in the chain.

[0123] As used herein, the term "alkylamino" refers to an amino group substituted by at least one alkyl group.

[0124] As used herein, the term "alkylthio" refers to a mercapto group substituted by an alkyl group, and can be represented by the general formula alkylS-.

[0125] As used herein, the term "amide" refers to the group

[0126]

[0127] wherein R 9and R 10 each independently represents hydrogen or a hydrocarbyl group, or R 9 and R 10 together with the N atom to which they are attached form a heterocyclic ring having 4-8 atoms in the ring structure.

[0128] The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines and salts thereof, for example, moieties which can be represented by:

[0129]

[0130] wherein R 9 , R 10 and R 10’ each independently represents hydrogen or a hydrocarbyl group, or R 9 and R 10 together with the N atom to which they are attached form a heterocyclic ring having 4-8 atoms in the ring structure.

[0131] As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group.

[0132] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group.

[0133] As used herein, the term "aryl" includes substituted and unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon. Preferably, the ring is a 5-7 membered ring, more preferably a 6 membered ring. The term "aryl" also includes polycyclic systems having two or more rings in which two or more carbons are common to two adjacent rings, wherein at least one ring is aromatic, for example, other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Aryl includes benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

[0134] The term "carbamate" is art-recognized and refers to the following group:

[0135]

[0136] wherein R 9 and R 10 independently represent hydrogen or a hydrocarbyl group.

[0137] As used herein, the term "carbocyclylalkyl" refers to an alkyl group substituted with a carbocyclyl group.

[0138] The term "carbocyclyl" includes 5-7 membered monocyclic and 8-12 membered bicyclic rings. Each ring of a bicyclic carbocyclyl can be selected from saturated, unsaturated, and aromatic rings. Carbocyclyl includes bicyclic molecules in which one, two, or three or more atoms are shared between two rings. The term "fused carbocyclyl" refers to a bicyclic carbocyclyl in which each ring shares two adjacent atoms with the other ring. Each ring of a fused carbocyclyl can be selected from saturated, unsaturated, and aromatic rings. In exemplary embodiments, an aromatic ring (e.g., phenyl) can be fused to a saturated or unsaturated ring, such as cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated, and aromatic bicyclic rings is included in the definition of carbocyclyl, where valence permits. Exemplary "carbocyclyl" groups include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene, and adamantane. Exemplary fused carbocyclyl groups include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-lH-indene, and bicyclo[4.1.0]hept-3-ene. A "carbocyclyl" can be substituted at any position(s) that can bear a hydrogen atom.

[0139] As used herein, the term "carbocyclylalkyl" refers to an alkyl group substituted with a carbocyclyl group.

[0140] The term "carbonate" is art-recognized and refers to the group -OCO2-.

[0141] As used herein, the term "carboxyl" refers to a group represented by the formula -CO2H.

[0142] As used herein, the term "ester" refers to a group -C(O)OR 9 wherein R 9 represents a hydrocarbyl group.

[0143] As used herein, the term "ether" refers to a hydrocarbyl group attached to another hydrocarbyl group through an oxygen. Thus, an ether substituent of a hydrocarbyl group can be hydrocarbyl-O-. The ether can be symmetrical or asymmetrical. Examples of ethers include, but are not limited to, heterocyclo-O-heterocyclo and aryl-O-heterocyclo. Ethers include "alkoxyalkyl," which can be represented by the general formula alkyl-O-alkyl.

[0144] As used herein, the terms "halo" and "halogen" mean halogen and include chlorine, fluorine, bromine, and iodine.

[0145] As used herein, the terms "hetaralkyl" and "heteroaralkyl" refer to an alkyl group substituted with a heteroaryl group.

[0146] The terms "heteroaryl" and "heteroaromatic" include substituted or unsubstituted aromatic monocyclic structures, preferably 5-7 membered rings, more preferably 5-6 membered rings, whose ring structure includes at least one heteroatom, preferably 1-4 heteroatoms, more preferably one or two heteroatoms. The terms "heteroaryl" and "heteroaromatic" also include polycyclic ring systems having two or more rings, where two or more carbons are common to two adjacent rings, where at least one ring is heteroaromatic, for example other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heteroaryl includes, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like.

[0147] As used herein, the term "heteroatom" means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0148] As used herein, the term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclyl group.

[0149] The terms "heterocyclyl," "heterocycle," and "heterocyclic" refer to substituted or unsubstituted non-aromatic ring structures, preferably 3-10 membered rings, more preferably 3-7 membered rings, whose ring structure includes at least one heteroatom, preferably 1-4 heteroatoms, more preferably one or two heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more rings, where two or more carbons are common to two adjacent rings, where at least one ring is heterocyclic, for example other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heterocyclyl includes, for example, piperidine, piperazine, pyrrolidine, morpholine, lactone, lactam, and the like.

[0150] As used herein, the term "hydrocarbyl" refers to groups that are bonded through carbon atoms that do not have =0 or =S substituents, and generally have at least one carbon-hydrogen bond and a predominantly carbon backbone, but can optionally include heteroatoms. Thus, for purposes of this application, groups such as methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered to be hydrocarbyl, but substituents such as acetyl (which has a =0 substituent on the linking carbon) and ethoxy (which is linked through oxygen rather than carbon) are not. Hydrocarbyl includes, but is not limited to, aryl, heteroaryl, carbocyclic, heterocyclic, alkyl, alkenyl, alkynyl, and combinations thereof.

[0151] As used herein, the term "hydroxylalkyl" refers to an alkyl group substituted with a hydroxyl group.

[0152] The term "lower" when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, means to include groups in which there are 10 or fewer atoms, preferably 6 or fewer atoms, in the substituent. For example, "lower alkyl" refers to an alkyl group containing 10 or fewer, preferably 6 or fewer, carbon atoms. In certain embodiments, the acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents defined herein are lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, respectively, whether they appear alone or in combination with other substituents, such as in the recitations of hydroxyalkyl and aralkyl (in which case, for example, atoms within the aryl group are not counted when counting the carbon atoms in the alkyl substituent).

[0153] The terms "polycyclyl," "polycyclic," and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl), for example, the rings are "fused rings," in which two or more atoms are common to two adjacent rings. Each ring in a polycyclic group can be substituted or unsubstituted. In certain embodiments, each ring in a polycyclic group contains 3-10 atoms, preferably 5-7.

[0154] The term "sulfate" is art-recognized and refers to the group -OSO3H or a pharmaceutically acceptable salt thereof.

[0155] The term "sulfonamide" is art-recognized and refers to a group represented by the general formula:

[0156]

[0157] wherein R 9 and R 10 independently represent hydrogen or a hydrocarbyl group.

[0158] The term "sulfoxide" is art-recognized and refers to the group -S(O)-.

[0159] The term "sulfonate" is art-recognized and refers to the group SO3H or a pharmaceutically acceptable salt thereof.

[0160] The term "sulfone" is art-recognized and refers to the group -S(O)2-.

[0161] The term "substituted" means a moiety having a substituent that replaces a hydrogen on one or more carbons of the backbone. It should be understood that "substitution" or "substituted with" includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is contemplated to include all allowed substituents of an organic compound. In a broad aspect, the allowed substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The allowed substituents can be one or more, and the same or different, for appropriate organic compounds. For purposes of this application, a heteroatom such as nitrogen can have a hydrogen substituent and / or any allowed substituent of an organic compound described herein that satisfies valence of the heteroatom. The substituents can include any of the substituents described herein, e.g., halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinite, amino, amido, amidine, imine, cyano, nitro, azido, thiol, alkylthio, sulfate, sulfonate, sulfamoyl, sulfamido, sulfonyl, heterocyclyl, aralkyl, or an aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that a moiety substituted on a hydrocarbon chain can itself be substituted, if appropriate.

[0162] As used herein, the term "thioalkyl" refers to an alkyl group substituted with a mercapto group.

[0163] As used herein, the term "thioester" refers to the group -C(O)SR 9 or -SC(O)R 9

[0164] wherein R 9 represents a hydrocarbyl group.

[0165] As used herein, the term "thioether" is equivalent to an ether wherein the oxygen is replaced with sulfur.

[0166] The term "urea" is art-recognized and can be represented by the general formula:

[0167]

[0168] wherein R 9 and R 10 independently represent hydrogen or a hydrocarbyl group.

[0169] As used herein, the term "modulate" includes inhibiting or suppressing a function or activity (such as cell proliferation) as well as enhancing a function or activity.

[0170] The phrase "pharmaceutically acceptable" is art-recognized. In certain embodiments, the term includes compositions, excipients, adjuvants, polymers and other materials and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0171] "Salt" is used herein to refer to an acid addition or base addition salt.

[0172] Many of the compounds useful in the methods and compositions of the present disclosure possess at least one stereogenic center in their structure. The stereogenic center can exist in the R or S configuration, the use of which is in accordance with the R / S conventions as described in Pure Appl. Chem. (1976), 45, 11-30. The present disclosure contemplates all stereoisomeric forms of the compounds, salts, prodrugs or mixtures thereof, including all possible mixtures of stereoisomers, such as enantiomeric and diastereomeric forms. See, e.g., WO 01 / 062726.

[0173] In addition, certain compounds containing alkenyl groups can exist as Z (syn) or E (anti) isomers. In each instance, both mixtures and individual isomers are included in the present disclosure.

[0174] Some compounds can also exist as tautomers. Such forms, although not explicitly indicated in the formulae depicted herein, are intended to be included within the scope of the present disclosure.

[0175] "Pharmaceutically acceptable" means approved or approvable by a regulatory agency of the Federal or state governments or the corresponding agency in countries other than the United States, or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0176] "Pharmaceutically acceptable salt" refers to a salt of the compound of the present invention that is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound. In particular, such salt is non-toxic and may be an inorganic or organic acid addition salt and a base addition salt. Specifically, such salts include: (1) acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; or with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid. Salts formed from sulfonic acid, camphor sulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-en-1-carboxylic acid, glucoheponic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthenic acid, salicylic acid, stearic acid, mucoconic acid, etc.; or (2) when the acidic protons present in the parent compound are replaced by metal ions (e.g., alkali metal ions, alkaline earth ions or aluminum ions), or when coordinated with organic bases such as ethanolamine, diethanolamine, triethanolamine, N-methylglucosamine, etc. By way of example only, salts further include sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, etc.; and when the compound contains basic functionality, it includes salts of non-toxic organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, methanesulfonate, acetate, maleate, oxalate, etc.

[0177] The term “pharmaceutically acceptable cation” refers to an acceptable cationic counterion with an acidic functional group. Examples of such cations include sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium cations, etc. (see, for example, Berge et al., J. Pharm. Sci. 66(1):1-79(1 / 77)).

[0178] "Pharmaceutically acceptable mediator" refers to a diluent, adjuvant, excipient, or carrier that is administered with the compounds of the present invention.

[0179] A “pharmaceutically acceptable metabolically cleavable group” is a group that cleaves in vivo to produce a parent molecule with the structure shown herein. Examples of metabolically cleavable groups include -COR, -COOR, -CONRR, and -CH2OR groups, wherein R in each case is independently selected from alkyl, trialkylsilyl, carbocyclic aryl, or carbocyclic aryl groups substituted with one or more of alkyl, halogen, hydroxyl, or alkoxy groups. Specific examples of representative metabolically cleavable groups include acetyl, methoxycarbonyl, benzoyl, methoxymethyl, and trimethylsilyl.

[0180] "Prodrug" means a compound which has a cleavable group and becomes by solvolysis or under physiological conditions an inventive compound that is pharmaceutically active in vivo, including a derivative of an inventive compound. Examples of such include, but are not limited to, choline ester derivatives and the like, N-alkylmorpholine esters and the like. Other derivatives of the inventive compounds, which exhibit activity in both their acid and acid derivative forms, can be prepared by modifying the inventive compounds in accordance with the techniques known to those of ordinary skill in the art. (See Bundgard, H., Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs include acid derivatives well known from the art, such as esters prepared by reaction of the parent acid with a suitable alcohol, or amides prepared by reaction of the parent acid compound with a substituted or unsubstituted amine, or acid anhydrides, or mixed anhydrides. Simple aliphatic or aromatic esters, amides and anhydrides derived from acidic groups pendent on the inventive compounds are particular prodrugs. In some cases it is desirable to prepare double ester type prodrugs, such as (acyloxy)alkyl esters or ((alkoxycarbonyl)oxy)alkyl esters. In particular, C1-C8alkyl, C2-C8alkenyl, C2-C8alkynyl, aryl, C7-C 12 substituted aryl and C7-C 12 arylalkyl esters of the inventive compounds.

[0181] "Solvate" means a form of a compound that typically involves the association of the compound with one or more solvent molecules, either through solvation or in the form of a hydration or alcoholation. Such physical associations involve varying degrees of ionic and / or covalent bonding, including hydrogen bonding. Typical solvents include water, ethanol, acetic acid, and the like. The compounds of the present application can be prepared, for example, in crystalline form and can be solvated or hydrated. Suitable solvates include pharmaceutically acceptable solvates, such as hydrates, and further include both stoichiometric solvates and non-stoichiometric solvates. In some cases, solvates will be capable of isolation, such as where one or more solvent molecules are incorporated into the crystal lattice of the solid state form of the compound. "Solvate" includes both solution-phase and isolatable solvates. Representative solvates include hydrates, ethanolates, and methanolates.

[0182] A "subject" to which administration is contemplated includes, but is not limited to, a human (i.e., a male or female of any age group, such as a pediatric subject (e.g., an infant, a child, an adolescent) or an adult subject (e.g., a young adult, a middle-aged adult, or an elderly adult)) and / or a non-human animal (e.g., a mammal, such as a primate (e.g., a cynomolgus monkey, a rhesus monkey), a bovine, a porcine, a equine, an ovine, a caprine, a rodent, a feline, and / or a canine). In certain embodiments, the subject is a human. In certain embodiments, the subject is a non-human animal. The terms "human," "patient," and "subject" are used interchangeably herein.

[0183] “Effective amount” means the amount of a compound that, when administered to a subject for treating or preventing a disease, is sufficient to effect such treatment or prevention. The “effective amount” can vary depending on the compound, the disease and its severity, and the age, weight, etc., of the subject to be treated. A “therapeutically effective amount” refers to an amount effective, when used in the treatment of a disease, to treat the disease. A “prophylactically effective amount” refers to an amount effective, when used in the prevention of a disease, to prevent the disease.

[0184] “Preventing” or “prevention” or “prophylactic treatment” refers to reducing the risk of acquiring or developing a disease or condition (i.e., causing at least one clinical symptom of the disease not to develop in a subject who is not yet exposed to a disease-causing agent or predisposed to the disease).

[0185] The term “prophylaxis” is related to “prevention” and refers to measures or procedures taken in the interest of preventing, rather than treating or curing, a disease. Non-limiting examples of prophylactic measures can include administration of a vaccine; administration of low molecular weight heparin to hospitalized patients at risk of thrombosis due to, for example, immobilization, and administration of an anti-malarial drug, such as chloroquine, prior to visiting a geographic region where malaria is endemic or the risk of contracting malaria is high.

[0186] “Treating” or “treatment” or “therapeutic treatment” of any disease or condition, in one embodiment, refers to ameliorating the disease or condition (i.e., arresting or reducing the manifestations, extent or severity of the disease or at least one clinical symptom thereof). In another embodiment, “treating” or “treatment” refers to improving at least one physical parameter that a subject can not discern. In still another embodiment, “treating” or “treatment” refers to modulating the disease or condition physically (e.g., stabilization of a discernible symptom), physiologically (e.g., stabilization of a physical parameter), or both. In a further embodiment, “treating” or “treatment” involves slowing the progression of the disease.

[0187] As used herein, the term “isotopic variant” refers to a compound that contains an unnatural proportion of an isotope at one or more of the atoms that constitute such compound. For example, an “isotopic variant” of a compound can contain one or more non-radioactive isotopes, such as deuterium ( 2 H or D), carbon-13 ( 13 C), nitrogen-15 ( 15 N), and the like. It will be appreciated that, in a compound in which such isotopic substitution is made, the following atoms, if present, can vary, such that, for example, any hydrogen can be “ 2 H / D, any carbon can be13 C, or any nitrogen can be 15 N, and the presence and location of such atoms can be determined within the skill of the art. Also, the present application can include isotopic variants of the compounds having radioisotopes, for example, where the resulting compound can be used in drug and / or substrate tissue distribution studies. The radioisotopes tritium (i.e. 3 H) and carbon-14 (i.e. 14 C) are particularly useful for this purpose due to their ease of incorporation and facile detection. Further, compounds substituted with positron emitting isotopes such as 11 C, 18 F, 15 O and 13 N can be prepared and used in positron emission tomography (PET) studies to examine substrate receptor occupancy. All isotopic variations of the compounds provided herein, whether radioactive or not, are intended to be encompassed within the scope of the present application.

[0188] It is also to be understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or in the spatial arrangement of their atoms, are termed "isomers." Isomers that differ in the arrangement of their atoms only in the spatial relationship are termed "stereoisomers."

[0189] Stereoisomers that are not mirror images of one another are termed "diastereomers" and those that are non-superimposable mirror images of each other are termed "enantiomers." Where a compound has an asymmetric center, for example, if it is attached to four different groups, a pair of enantiomers is possible. An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and is designated as either dextrorotatory or levorotatory (i.e., as (+)- or (-)-isomers). Chiral compounds can exist as individual enantiomers or as mixtures of its enantiomers. A mixture containing equal proportions of each enantiomer is called a "racemic mixture".

[0190] "tautomers" refer to compounds that are interchangeable forms of a particular chemical structure, and differ only in the arrangement of electrons and atoms, usually hydrogen and electrons. Thus, two structures can be in equilibrium by movement of their electrons and atoms, usually H. For example, enol and ketone are tautomers because they rapidly interconvert with one another by treatment with an acid or base. Another example of tautomerism is the acid (aci-) and nitro form of phenylnitromethane, which likewise form by treatment with an acid or base. The tautomeric form can be relevant to achieving optimal chemical reactivity and biological activity of the subject compound.

[0191] As used herein, a pure enantiomeric compound is substantially free of other enantiomers or stereoisomers of the compound (i.e., an enantiomeric excess). In other words, the "S" form of a compound is substantially free of the "R" form of the compound and thus is an enantiomeric excess of the "R" form. The term "enantiomerically pure" or "pure enantiomer" means that the compound contains greater than 95% by weight, greater than 96% by weight, greater than 97% by weight, greater than 98% by weight, greater than 98.5% by weight, greater than 99% by weight, greater than 99.2% by weight, greater than 99.5% by weight, greater than 99.6% by weight, greater than 99.7% by weight, greater than 99.8% by weight, or greater than 99.9% by weight of an enantiomer. In certain embodiments, the weight is based on the total weight of all enantiomers or stereoisomers of the compound.

[0192] As used herein, and unless otherwise specified, the term "enantiomerically pure R- compound" means at least about 95% by weight of the R-compound and at most about 5% by weight of the S-compound, at least about 99% by weight of the R-compound and at most about 1% by weight of the S-compound, or at least about 99.9% by weight of the R-compound and at most about 0.1% by weight of the S-compound. In certain embodiments, the weight is based on the total weight of the compound.

[0193] As used herein, and unless otherwise specified, the term "enantiomerically pure S- compound" or "S-compound" means at least about 95% by weight of the S-compound and at most about 5% by weight of the R-compound, at least about 99% by weight of the S-compound and at most about 1% by weight of the R-compound, or at least about 99.9% by weight of the S-compound and at most about 0.1% by weight of the R-compound. In certain embodiments, the weight is based on the total weight of the compound.

[0194] In the compositions provided herein, an enantiomerically pure compound or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof can be present with other active or inactive ingredients. For example, a pharmaceutical composition comprising an enantiomerically pure R-compound can comprise, for example, about 90% excipient and about 10% enantiomerically pure R-compound. In certain embodiments, an enantiomerically pure R-compound in such a composition can comprise, for example, at least about 95% by weight R-compound and at most about 5% by weight S-compound, based on the total weight of the compound. For example, a pharmaceutical composition comprising an enantiomerically pure S-compound can comprise, for example, about 90% excipient and about 10% enantiomerically pure S-compound. In certain embodiments, an enantiomerically pure S-compound in such a composition can comprise, for example, at least about 95% by weight S-compound and at most about 5% by weight R-compound, based on the total weight of the compound. In certain embodiments, the active ingredient can be formulated with little or no excipient or carrier.

[0195] The compounds of the present application can have one or more asymmetric centers; thus, such compounds can be produced as individual (R)- or (S)-stereoisomers, or as mixtures of them.

[0196] Unless otherwise specified, a description or naming of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures (racemic or otherwise) thereof. Methods on the determination of stereochemistry and separation of stereoisomers are well-known in the art.

[0197] One of ordinary skill in the art of organic synthesis will recognize that the maximum number of heteroatoms in a stable, chemically feasible heterocycle (whether it is aromatic or non-aromatic) is determined by the size of the ring, the unsaturation, and the valence of the heteroatom. Generally, a heterocycle can have 1-4 heteroatoms, provided that the heteroaromatic ring is chemically feasible and stable.

[0198] Examples

[0199] In order that the application described herein can be more fully understood, the following examples are set forth. The examples described in this application are offered to illustrate the compounds, compositions, materials, devices, and methods provided herein and are not to be construed in any way as limiting their scope.

[0200] Materials and Methods

[0201] General:

[0202] All chemicals for lipid synthesis were purchased from Sigma-Aldrich and used as received. All ASOs and DNA fragments were purchased from Integrated DNA Technologies (IDT). ASOs were used as provided by IDT, and when noted, we used ASO products provided by the company to include chemical modifications for improved stability. HeLa-DsRed and GFP-HEK cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM, Sigma-Aldrich) supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich) and 1% penicillin-streptomycin (Gibco). Fluorescence intensity of GFP-HEK cells was analyzed by flow cytometry (BD FACS Calibur, BD Science, CA). GFP-Cre (addgene #89253) protein was expressed and extracted from BL21 E. coli and further purified by Ni-NTA column (Qiagen). Nanoparticle size and zeta potential were recorded on ZetaPALS particle size analyzer. TEM images were taken by FEI Technai Spirit transmission electron microscope.

[0203] Lipid synthesis

[0204] All head groups for lipid synthesis were commercially available from Sigma-Aldrich. All cationic lipids (NT1-012B~018B, NT2-012B~018B, NT3-012B~018B, NT1-EC16, NT1-C18, NT1-1E, NT2-EC16, NT2-1E, NT3-EC16, NT3-1E, 306-012B-3, 76-016B) were synthesized according to our previous reports. Crude products were purified by flash chromatography on silica gel. 1E tail was synthesized as shown in Figure 18A Figure 18B Figure 18C

[0205] Biodistribution of DiR-labeled NT-LNP in mouse brain

[0206] ​​​NT-based lipids and DiR were dissolved together in 100% ethanol at a weight ratio of 10: 1. Then, 100 pL of the solution was added dropwise to 300 pL of sodium acetate buffer (25 mM, pH 5.2) and vortexed briefly. Finally, we removed the ethanol from the formulation by dialysis (MWCO 35 kDa, ThermoFisher) against diH20 for 12 hours. The DiR-labeled LNP was then injected intravenously into BALB / C mice (female, 6 weeks old). After 1 hour, the mice were anesthetized and perfused with normal saline. After that, the mice’s brains were collected. The fluorescence signal distribution was visualized using a Spectrum CT biophotonic imager (PerkinElmer, Boston, MA).

[0207] Preparation of AmB / NT-lipid nanoparticle formulations

[0208] AmB encapsulates were prepared according to our previous report. Briefly, 1 mg of each lipid (solid) was mixed with 1 mg of AmB in 300 pL of dimethyl sulfoxide (DMSO). The mixture was sonicated for 30 minutes and then vortexed for 10 minutes until completely dissolved. The solution was added dropwise to a glass vial containing 600 pL of sodium acetate buffer (pH 5.0) for homogenization at 700 rpm. The solution was further dialyzed against distilled water overnight using a dialysis tube (MWCO 35 kDa) to remove DMSO and unencapsulated AmB.

[0209] Characterization of AmB / NT-lipid nanoparticle formulations

[0210] The particle size and polydispersity index (PDI) of all encapsulates were measured using dynamic light scattering (DLS). The zeta potential was recorded on a ZetaPALS particle size analyzer. The DLC of AmB was calculated according to our previous report. TEM images were taken by a FEI Technai Spirit transmission electron microscope.

[0211] Data analysis

[0212] Data analysis was performed using one-way analysis of variance (ANOVA) followed by Turkey-Kramer’s multiple comparison test for more than two groups. Prism (v.8, GraphPad Software, La Jolla, CA) was used to compare two groups using the Student’s t-test. Values of P < 0.05 were considered significant.

[0213] NT-lipid synthesis and BBB permeability study of NT-LNP

[0214] Neurotransmitters tryptophan, phenylethylamine, and phenylethanolamine were selected as the structural basis for the synthesis of lipids. NT lipids were synthesized via Michael addition between the primary amine of the neurotransmitter and its hydrophobic tail containing an acrylate in a glass vial at 70°C for 48 hours. Figure 1 B) A method similar to our previously published strategy for synthesizing combinatorial lipid libraries was used. The result was a combinatorial library of NT-lipids, each containing a specific neurotransmitter as the head and a specific bioreducible hydrophobic structure as the tail. This NT-lipid was named “NTn-O[x]B” (n = 1, 2, 3), where NT1 is tryptophan, NT2 is phenylethylamine, NT3 is phenylethanolamine, and O[x]B represents the bioreducible hydrophobic tail, where [x] indicates… Figure 1 B indicates the number of carbon atoms in the hydrophobic tail of the acrylate. For example, NT1-O12B represents a lipid containing a tryptophan head group and a hydrophobic tail containing 12 carbon atoms. All NT lipids were purified using rapid chromatography and characterized by ESI-MS. Figure 5 The resulting NT-lipids are amphiphilic and therefore capable of self-assembling into micelles or liposomes when prepared in aqueous solution. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) of the NT-lipids confirmed that these structures indeed self-assemble into spherical liposome structures. Figure 6 ).

[0215] Table 1. MS values ​​of synthesized NT-derived lipids

[0216]

[0217]

[0218]

[0219]

[0220] Using a fluorescent dye (DiR) as a model cargo, this study further investigated whether these NT-lipids could cross the blood-brain barrier (BBB) ​​during systemic intravenous delivery. Hydrophobic small molecules, such as DiR, can partition into the hydrophobic regions of micelles and liposomes and have been frequently used to track the biodistribution of these structures. To formulate DiR-loaded NT-lipids, NT-lipids and DiR were mixed in ethanol at a 10 / 1 (w / w) ratio. The mixture was then added dropwise to sodium acetate buffer (25 mM, pH 5.2), and the ethanol was removed by dialysis. The DiR-loaded NT-lipid nanoparticle solution was injected into mice via tail vein injection. One hour later, the animals were euthanized and perfused with saline. The skulls were removed, and the brains were imaged using an IVIS imaging device (PerkinElmer) at an excitation wavelength of 750 nm.

[0221] As Figure 1 C, a strong DiR fluorescence signal was observed in the brain of mice treated with DiR / NT1-lipidoid nanoparticles, in contrast to the very weak fluorescence signal observed in the brain of mice treated with DiR / NT2-lipidoid and DiR / NT3-lipidoid. It was also observed that the length of the aliphatic tail significantly affected the observed fluorescence intensity, with NT1-lipidoids containing shorter aliphatic tail lengths leading to greater fluorescence intensity Figure 7 ). There were no significant differences between these NT1-derived lipidoids in physical properties such as hydrodynamic size, polydispersity index, zeta potential and morphology Figure 6

[0222] It was hypothesized that doping NT1-lipidoids, such as NT1-012B, into other BBB-impermeable lipid formulations would result in the resultant lipid formulation crossing the BBB. Previously published synthetic lipids 76-016B, EC16-80 and 113-016B were used to test this ability to deliver DiR to the brain. It was found that none of these lipids could effectively deliver DiR into the mouse brain by themselves, however, upon doping NT1-012B into these lipids, a strong DiR signal was observed in the mouse brain Figure 8

[0223] The chemical structure of NT1 is based on the neurotransmitter dimethyltryptamine, which has been reported to cross the BBB by active transport across the endothelial cell plasma membrane. 21 It was hypothesized that our results were also driven by active transport, and that changes in the chemical structure of the NT1 lipid would modulate its ability to cross the BBB. We specifically hypothesized that the ionizability of the alpha-amine in the lipified tryptamine (NT1) was an important factor in the ability of the derivative to cross the BBB. To test this hypothesis, a series of NT1 derivatives with different linkers were synthesized, as shown in Figure 9 DiR signal was observed from the brains of mice treated with all NT1 derivatives except NT1-neu. In NT1-neu, the alpha-amine in the tryptamine was connected by an amide bond, which is not ionizable, whereas the alpha-amine in all other NT1 derivatives was ionizable. Furthermore, no strong DiR signal was observed from the brains of mice treated with NT2 and NT3 derived lipidoids with any linker.

[0224] Delivery of small molecule AmB into mouse brain

[0225] ​​As shown above, NT1 -derived lipids were identified to be able to deliver hydrophobic dye (DiR) into the brain, whether used alone or doped into other LNPs. These NT1 -derived lipids were used to examine the delivery of a therapeutically relevant hydrophobic drug molecule into the brain. Amphotericin B (AmB) was chosen as the model drug. AmB is a classic polyene antifungal drug and is the gold standard for treating severe systemic fungal infections. However, due to its BBB impermeability, it cannot be used clinically to treat brain fungal infections. Recently, AmB was formulated in synthetic lipid nanoparticles and a comprehensive PK and biodistribution study was performed on the AmB formulation using traditional synthetic lipid nanoparticles, but in that study, none of our lipid nanoparticles were able to penetrate the BBB to deliver AmB into the mouse brain 27 .

[0226] AmB was encapsulated in pure NT1 -lipids (i.e. NT1 -O12B, NT1 -O14B, NT1 -O16B and NT1 -O18B) using a procedure similar to DiR encapsulation. Mice were injected intravenously via the tail vein with AmB-loaded NT1 -lipid nanoparticles at a dose of 5 mg / kg AmB per mouse. After 24 hours, animals were sacrificed and brains were harvested, perfused with normal saline and homogenized. AmB concentration in brain tissue was quantified using HPLC (detailed method in SI). As shown in Figure 10 , the concentration of AmB in brain tissue for all four groups was approximately 150 ng / g tissue. Notably, in our previous report, AmB was not detectable in the brain after systemic delivery with traditional synthetic lipids, suggesting that the NT1 -lipid formulation enhanced the delivery of AmB into the mouse brain.

[0227] However, AmB formulated in NT-lipid formulations appeared as opaque solutions Figure 11 A), indicating large particle size in solution. DLS results Figure 11 B, Figure 12 showed that the diameter of the nanoparticles ranged from 750-800 nm. It was hypothesized that making the NT1 -lipid nanoparticles smaller could help improve the delivery efficiency to the brain. In a previous report, it was found that quaternized lipids provided stable AmB formulations with smaller particle sizes compared to non-quaternized lipids. 27 Therefore, it was hypothesized that doping NT1 -lipids in quaternized lipids could result in smaller nanoparticle size while maintaining or improving the ability to penetrate the BBB.

[0228] A new phenylboronic acid quaternized lipid, PBA-Q76-O16B Figure 2A) for Amb encapsulation. NT1-012B was chosen as the dopant to enhance brain delivery because it showed the highest DiR fluorescence intensity among all NT-based lipids Figure 1 C). AmB was formulated in a mixture of NT1-012B and PBA-Q76-016B, both lipids were mixed at different weight ratios (7:3, 5:5, 3:7, 1:9 and pure PBA-Q76-016B). As shown in Figure 2 B, with the increase of PBA-Q76-016B lipid percentage in the formulation, the AmB encapsulates gradually turned into a uniform transparent yellow solution. The hydrodynamic size also decreased from 800 nm to 100 nm Figure 2 C, Figure 12 ) Using DiR as cargo, we observed that the lipids containing NT1-012B and PBA-Q76-016B at a 3:7 (w / w) ratio provided the strongest fluorescence signal in the mouse brain when compared to all other lipid ratios Figure 2 D). The fluorescence signal intensity at 3:7 ratio was 4.5 times higher than that of the brain treated with DiR formulated in pure NT1-012B Figure 13 ) Further investigation was carried out on AmB delivery using mixed lipids and the AmB concentration in mouse brain tissues was determined 24 hours after intravenous injection of 5 mg / kg AmB / mouse. As shown in Figure 2 E, with the increase of PBA-Q76-016B doping ratio from 0% (i.e. pure NT1-012B) to 70% (i.e. 3:7 ratio), the amount of AmB detected in the brain increased and reached a maximum concentration of about 300 ng / g, which was about 2 times higher than that of pure NT1-012B. When the doping ratio was further increased to 90% (i.e. 1:9), the AmB concentration was slightly lower, but still higher than that treated with AmB formulated in pure NT1-012B. Therefore, the results of AmB delivery closely matched those of DiR delivery Figure 2 D and 2E). Interestingly, in the absence of doping NT1-lipids, almost no Amb was detected in the brain after intravenous injection of pure PBA-Q76-016B / Amb. These results indicate the key role of NT1 lipids in facilitating brain delivery and the importance of finding the optimal doping ratio.

[0229] Delivery of nucleic acid Tau-ASO into mouse brain for gene knockdown

[0230] The efficiency of mixed lipid formulations for in vitro delivery of ASO was evaluated by delivering ASO targeting GFP mRNA Figure 3 B) to HEK cells stably expressing green fluorescent protein (GFP). NT1-014B alone did not show GFP silencing effect Figure 3The 10:0 ratio in B indicates that this lipid alone is ineffective for intracellular ASO delivery. However, GFP silencing was observed when ASO was delivered using LNP containing a mixture of NT1-O14B and 306-O12B-3. GFP silencing was observed in GFP-HEK cells when the doping ratio of 306-O12B-3 was greater than 50% (i.e., a 5:5 weight ratio or more favorable to 306-O12B-3), and the silencing efficiency increased with increasing 306-O12B-3 doping ratio. Disrupted ASO delivered by Lipofetamine 2000 (LPF 2K) did not show GFP silencing, indicating that GFP silencing is indeed ASO sequence-specific.

[0231] The study then investigated whether a mixed lipid formulation (NT1-O14B and 306-O12B-3) could deliver ASO to the brain and mediate gene knockdown in vivo. Tau was chosen as the therapeutic target, and an ASO targeting tau mRNA was engineered because ASO-mediated tau reduction has shown promising results in treating Alzheimer's disease (AD) following local injection of Tau-ASO using an intraventricular (ICV) pump. 31,32 .

[0232] The Tau-ASO sequence was selected based on publicly available literature. 31 The Tau-ASO provided is chemically modified with phosphate thiocyanate groups between each nucleic acid and 2'-O-methoxyethyl groups at the 5' and 3' ends of the ribose to enhance efficacy. To formulate the ASO for intravenous injection, the ASO was mixed with the prepared LNP solution at a weight ratio of 1 / 15 (ASO to total lipids). Each mouse received five injections of 1 mg / kg ASO, spaced three days apart. Mice were sacrificed four days after the last injection, perfused, and brain tissue was harvested and homogenized to extract total RNA. Total tau mRNA levels were analyzed by quantitative PCR. Figure 3 As shown in Figure C, no reduction in tau mRNA in brain tissue was detected when ASO was delivered using pure NT1-O14B or pure 306-O12B-3. For mixed lipid formulations, only NT1-O14B and 306-O12-3 at w / w ratios of 5:5 and 3:7 showed a reduction in tau mRNA in the brain. These two formulations resulted in approximately 25% and 50% mRNA reductions, respectively. No tau mRNA silencing was observed in mixed lipid formulations at other ratios (i.e., 7:3 and 1:9).

[0233] To confirm that ASO delivery leads to functional knockdown of tau, we also examined tau protein levels in ASO-treated mice using ELISA. Figure 3D). Mice treated with Tau-ASO formulated in NT1-O14B / 306-O12B-3 (3:7 w / w) showed a strong reduction in total tau protein levels compared to the untreated group. Furthermore, using the exact same method as for the functional ASO, the perturbing Tau-ASO was delivered in the best performing ratio (NT1-O14B / 306-O12B-3 at 3:7 w / w). As shown, no tau mRNA silencing effect was detected nor a reduction in tau protein, indicating that the tau knockdown was specifically due to sequence-specific ASO silencing.

[0234] Delivery of GFP-Cre fusion protein for gene recombination in Ai14 mouse brain The GFP-fused Cre recombinase was chosen as model protein for the study, using the Ai14 model mouse strain Figure 4 A). The Ai14 mouse strain contains a flox-stop-flox tdTomato construct. Successful intracellular delivery of Cre protein into the cells of the Ai14 mouse leads to genetic recombination and turns on the expression of tdTomato, which can be directly visualized as a red fluorescent signal without additional staining. Here the (-27)GFP-Cre protein was used. NT1-O14B LNP doped with PBA-Q76-O16B were chosen because these nanoparticles can successfully deliver (-27)GFP-Cre. Based on the results observed from the delivery of AmB and ASO, the weight ratio of NT1-O14B and PBA-Q76-O16B was fixed at 3:7. The lipid formulation was prepared using the method described for the ASO delivery formulation. Briefly, the (-27)GFP-Cre protein was mixed with the LNP at a weight ratio of 1 / 4 and the solution was incubated at room temperature for 15 minutes before intravenous injection. Mice were injected four times with a dose of 50 pg protein per injection. Five days after the last injection, mice were sacrificed and brain tissue was collected, fixed and dehydrated. The tissue was then cryosectioned into 15 pm slices and counterstained with DAPI for fluorescence imaging. As shown in Figure 4 B, strong tdTomato signal was observed in multiple regions of the brain, including the cerebral cortex, hippocampus and cerebellum. In contrast, no tdTomato expression was observed in the brains of mice injected with LNP formulations using pure NT1-O14B (10:0) or pure PBA-Q76-O16B (0:10).

[0235] Different formulations of GFP-Cre fusion protein for gene recombination in Ai14 mouse brainNT1-O14B was doped into various lipidoid nanoparticle formulations, including 306-O12B, PBA-Q76O16B, Dlin-MC3, to investigate the doped LNP formulations for delivery of Cre mRNA into the brain of Ai14 mice by intravenous injection. The weight ratio of NT1-O14B and other ionizable lipids (e.g. 306-O12B, PBA-Q76O16B, Dlin-MC3) was 3:7. To formulate stable LNPs, other helper lipids were also included, including (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG2000)), cholesterol, and DOPE. mRNA encoding Cre recombinase was loaded into the LNPs and injected into Ai14 mice. Mice were sacrificed at specific time points and brain tissues were collected, fixed and dehydrated. The tissues were then frozen sectioned into 15 pm sections and counterstained with DAPI for fluorescent imaging. tdTomato signal was observed in multiple regions of the brain, indicating successful delivery of Cre mRNA into brain cells by systemic injection with such LNP formulations. Figures 19A-19N 、 Figures 20A-20B and Figures 21A-21B Fluorescent images of Ai14 mouse brain sections are shown in FIGS. 1-3. 306-O12B doped with NT1-O14B showed the highest brain delivery compared to PBA-O76O16B or Dlin-MC3 LNPs.

[0236] Cited References

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[0266] https: / / doi.org / 10.1016 / j.omtn.2020.01.018

[0267] 31. De Vos, S. L. et al. Antisense reduction of tau in adult mice protects against seizures. J. Neurosci. 33, 12887-12897 (2013).

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[0269] incorporated by reference

[0270] All of the U.S. and PCT patent publications and U.S. patents referred to herein are hereby incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In the event of a conflict between the present application (including any definitions herein) and any incorporated publication or patent, the present application (including any definitions herein) shall control.

[0271] Other embodiments

[0272] Those skilled in the art will recognize or be able to ascertain using not more than routine experimentation many equivalents to the specific embodiments described herein. The scope of the present embodiments described herein is not intended to be limited to the above description but rather is to be accorded the broadest interpretation of the appended claims to encompass all equivalent combinations of features. Those skilled in the art will appreciate that various adaptations and modifications of the described embodiments can be configured without departing from the scope and spirit of the application as defined by the appended claims.

Claims

1. A compound of Formula I: Formula I or a pharmaceutically acceptable salt thereof, wherein: (I)， Z is -CH2-, -O-, or -S-; Y is W is -NR 20 -; R 脂质 For , wherein: R 1 and R 2 are independently -H, -OH, or -SH; R 3 and R 4 are both -H; or R 3 and R 4 together form an oxo (=0) group; m is an integer selected from 1-3; X and Y 1 independently -CH2-, -O-, or -S-; n is an integer selected from 1-14; p is 0 or 1; q is an integer selected from 1-10; t is 0 or 1; and 7. The compound of claim 1, wherein Z is -CH2- or -O-. R 20 For R 脂质 .

2. The compound of claim 1, wherein R 1 and R 2 are independently -H or -OH.

3. The compound of claim 1, wherein R 1 and R 2 are -H.

4. The compound of claim 1, wherein R 1 is -H; and R 2 is -OH.

5. The compound of claim 1, wherein R 3 and R 4 are -H.

6. The compound of claim 1, wherein R 3 and R 4 together form an oxo (=0) group.

8. The compound of claim 7, wherein Z is -CH2-.

9. The compound of claim 7, wherein Z is -O-.

17. The compound of claim 1, wherein m is 1 or 2.

10. The compound of claim 1, wherein R 1 and R 2 are -H, R 3 and R 4 together form an oxo (=0) group, and Z is O.

11. The compound of claim 1, wherein R 1 is -H, R 2 is -OH, R 3 and R 4 is -H, and Z is -CH2-.

12. The compound of claim 1, wherein X and Y 1 are independently -CH2- or -O-.

13. The compound of claim 12, wherein X and Y are independently -CH2- or -O- with the proviso that X and Y are not the same. 1 are independently -CH2- or -O-, with the proviso that X and Y are not the same.

14. The compound of claim 1, wherein X and Y 1 are independently -CH2- or -S-.

15. The compound of claim 14, wherein X and Y 1 are both -CH2-.

16. The compound of claim 14, wherein X and Y 1 are both -S-.

18. The compound of claim 17, wherein m is 1.

19. The compound of claim 17, wherein m is 2.

20. The compound of claim 1, wherein n is an integer selected from 4-12.

21. The compound of claim 20, wherein n is an integer selected from 6-10.

22. The compound of claim 1, wherein p is 0.

23. The compound of claim 1, wherein p is 1.

24. The compound of claim 1, wherein q is an integer selected from 2-8.

25. The compound of claim 24, wherein q is an integer selected from 4-8.

26. The compound of claim 1, wherein t is 0.

27. The compound of claim 1, wherein t is 1.

28. A compound selected from: and or a pharmaceutically acceptable salt thereof. 、 29. A lipoid nanoparticle comprising the compound of any one of claims 1-28. , 30. The lipoid nanoparticle of claim 29, further comprising a protein.

31. The lipoid nanoparticle of claim 30, wherein the protein is GFP-Cre.

32. The lipoid nanoparticle of claim 29, further comprising a nucleic acid.

33. The lipoid nanoparticle of claim 32, wherein the nucleic acid is a Tau-ASO.

34. The lipoid nanoparticle of claim 29, further comprising a small molecule.

35. The lipoid nanoparticle of claim 34, wherein the small molecule is an antifungal agent or a chemotherapeutic agent. ​ ​ 36. The lipidoid nanoparticle of claim 34, wherein the small molecule is selected from the group consisting of bortezomib, imatinib, gefitinib, erlotinib, afatinib, osimertinib, dacomitinib, daunorubicin hydrochloride, cytarabine, fluorouracil, irinotecan hydrochloride, vincristine sulfate, methotrexate, paclitaxel, epirubicin, docetaxel, cyclophosphamide, carboplatin, lenalidomide, ibrutinib, abiraterone acetate, enzalutamide, pemetrexed, palbociclib, nilotinib, everolimus, lucotinib, pirarubicin, idarubicin, valrubicin, amrubicin, bleomycin, patulin, dactinomycin, plicamycin, streptozotocin, pentostatin, the mytomycin class of mitomycin C, the enediyne class of calicheamicin, the glycoside class of duocarmycin, the macrolide class of epothilone, ixabepilone, salinosporamide A, vinblastine, vincristine, etoposide, teniposide, vinorelbine, camptothecin, belinostat, cryptophycin, thailanin, trabectedin, aplidine, and ecteinascidin 743 (ET743).

37. The lipidoid nanoparticle of claim 34, wherein the small molecule is amphotericin B or doxorubicin.

38. The lipidoid nanoparticle of claim 29, wherein the lipidoid nanoparticle has a particle size of 25 nm to 1000 nm.

39. The lipidoid nanoparticle of claim 38, wherein the lipidoid nanoparticle has a particle size of 50 nm to 500 nm.

40. A pharmaceutical composition comprising the lipidoid nanoparticle of claim 29; and a pharmaceutically acceptable carrier or excipient.

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