Synthetic lipid-like materials for brain delivery

Abstract

Disclosed are (i) a compound of formula I or a pharmaceutically acceptable salt thereof; and (ii) lipidoid nanoparticles comprising a compound of formula I or a pharmaceutically acceptable salt thereof; and their use as vehicles for drug delivery across the blood-brain barrier.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 019,530, filed May 4, 2020, the contents of which are incorporated herein by reference in their entirety. GOVERNMENT SUPPORT

[0002] This invention was made with government support under Grant Nos. TR002636 and EB027170 awarded by the National Institutes of Health. The government has certain rights in this invention. [Background technology]

[0003] Treatment of central nervous system (CNS) diseases, such as neurodegenerative disorders, brain tumors, brain infections, and stroke, is severely constrained by the blood-brain barrier (BBB), which prevents the transport of most small molecule drugs and macromolecules (e.g., peptides, gene drugs, and protein drugs) into the brain. To date, extensive efforts have been made to enhance brain delivery efficiency, including direct CNS administration, BBB disruption, and carrier vehicle-mediated delivery. However, direct CNS administration is invasive, can cause infection and tissue damage, and is also limited by diffusion distances and the rapid efflux of drugs from the CNS within a few hours. Disruption of the BBB using techniques such as osmotic disruption, biochemical disruption, and ultrasound-mediated disruption is effective in introducing drugs into the brain, but these transient BBB openings also allow leakage of plasma proteins into the brain, leading to neurotoxicity, vascular lesions, and chronic neuropathological changes in the brain. Therefore, approaches for the safe and efficient delivery of BBB-impermeable cargoes to the CNS, particularly for gene and nucleic acid therapies, remain desirable.

[0004] Carrier vehicle-mediated brain drug delivery is considered a promising and versatile brain delivery system. 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 gene delivery to the brain, but have limitations such as production costs and safety concerns. Due to their non-immunogenicity, exosomes have been utilized to deliver small molecules, proteins, and nucleic acids to the brain; however, many challenges remain in isolation methods, cargo loading procedures, in vivo toxicity, and pharmacokinetics. The molecular Trojan horse approach, which relies on receptor-specific monoclonal antibodies or peptides to deliver genetically fused cargoes to the brain, shows promise for delivering biological substances across the BBB. However, the manufacturing process must be specifically tailored for each different biological cargo, and its stability, safety, and immunogenicity pose challenges for clinical development. Crossing the BBB using various nanoparticles, such as liposomes, cationic polymers, inorganic nanoparticles, and nanocapsules, has shown promise in the delivery of various cargoes to the CNS, but complex modifications are always required to ensure that the produced particles are BBB-permeable.

[0005] Neurotransmitters are endogenous chemicals that enable neurotransmission. In particular, several neurotransmitters have been demonstrated to cross the BBB. For example, dimethyltryptamine and other tryptamine derivatives have been shown to cross the BBB by active transport across the endothelial cell membrane. Summary of the Invention [Means for solving the problem]

[0006] Disclosed herein is a simple and effective approach for delivering cargo to the brain using neurotransmitter-derived synthetic lipids. This approach is highly robust and can be used to successfully deliver different classes of cargo (such as small molecules, nucleic acids, and proteins), all using the same simple nanoparticle design.

[0007] In one aspect, a compound of the following formula or a pharmaceutically acceptable salt thereof is disclosed: [ka] [In the formula, Y is a moiety derived from a neurotransmitter; W is -NR 20 -, -O-, or -S-; R 脂質 are 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 is heteroalkynyl; R 20 is R 脂質 , H., C. 1-6 Alkyl, C 1-6 Alkenyl, or C 1-6 alkynyl].

[0008] In certain aspects, lipidoid nanoparticles comprising the compounds described herein are disclosed.

[0009] In certain aspects, disclosed are pharmaceutical compositions comprising the lipidoid nanoparticles described herein and a pharmaceutically acceptable carrier or excipient. [Brief explanation of the drawings]

[0010] [Figure 1A]FIG. 1 is a schematic diagram of the formulation of NT-lipidoid-doped LNPs for cargo delivery to the brain. [Figure 1B] FIG. 1 is a schematic diagram of the synthetic pathway, lipid nomenclature, and chemical structures of neurotransmitters used in lipidoid synthesis. [Figure 1C] Representative ex vivo fluorescence images of a dissected brain 1 hour after a single intravenous injection of 1 mg / kg DiR-labeled NT-LNPs. DiR was doped into the NT-LNPs at a weight ratio of 10%. Mice were perfused with saline before dissection. [Figure 2A] 1A-1C are chemical structures of PBA-Q76-O16B, NT1-O12B, and a schematic diagram of the doped NT1-lipidoid AmB formulation. [Figure 2B] Photographs of AmB formulations in NT1-O12B (using weight ratios) doped with different amounts of PBA-Q76-O16B. Pure NT1-O12B / AmB inclusion bodies appeared as an opaque suspension, but the appearance of the inclusion bodies changed from a translucent solution to a homogeneous, transparent yellow solution as the doping ratio of PBA-Q76-O16B lipidoid increased. [Figure 2C] 1 is a graph showing the hydrodynamic diameter and polydispersity index of different NT-LNP / AmB formulations as determined by DLS measurements. [Figure 2D] Representative fluorescence image of a mouse dissected brain 1 h after a single intravenous injection of 1 mg / kg DiR-loaded NT1-O12B / PBA-Q76-O16B LNPs. The weight ratio of DiR in the LNPs is 10%. [Figure 2E] This graph shows AmB concentrations in brain tissue 24 hours after intravenous injection of 5 mg / kg AmB in various NT1-O12B / PBA-Q76-O16B LNP formulations (n=4 / group) measured using HPLC. Mice were perfused with saline and then dissected. One-way ANOVA, Sidak post hoc analysis, *p<0.05, **p<0.001, or ***p<0.0001. Graph data are presented as boxplots with individual points overlaid; error bars represent maximum and minimum values; and the line within the box represents the median. [Figure 3A] Chemical structures of 306-O12B-3, NT1-O14B, and a schematic diagram of the doped NT-lipidoid tau-ASO formulation for brain delivery are shown. [Figure 3B] Figure 1 shows the GFP silencing efficiency of HEK-GFP cells treated with or without ASO / NT-LNP complexes. NT1-O14B LNPs alone showed no silencing efficacy, but doping 306-O12B-3 LNPs with NT1-lipidoid resulted in successful gene silencing in vitro. *p<0.01 vs. all other samples in the same group. [Figure 3C] (Figure 1) shows that tau-ASOs (Tau-ASOs) formulated with NT1-O14B, saline, or scrambled tau-ASO-LNPs doped with different ratios of 306-O12B-3 were intravenously injected via the tail vein into C57BL / 6J mice (n=6 / group), and brains were analyzed for total tau mRNA levels. Graph data are presented as boxplots with individual points overlaid, error bars represent maximum and minimum values, and lines within boxes represent median values. *p<0.05 or **p<0.001.) [Figure 3D] Graph showing total tau protein levels in the NT1-O14B / 306-O12B-3=3:7 group compared to saline or scrambled tau-ASO. **p<0.001, One-way ANOVA, Sidak post-hoc analysis. [Figure 4A] FIG. 1 is a schematic diagram of a mixed LNP formulation using NT1-O14B and PBA-Q76-O16B for delivery of GFP-Cre protein to the brain. [Figure 4B]Fluorescence images 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 NT1-O14B / PBA-Q76-O16B LNPs at a ratio of 3:7, 10:0, or 0:10. After 3 weeks, the NT1-O14B / PBA-Q76-O16B 3:7 group showed tdTomato expression in the cerebral cortex, hippocampus, and cerebellum, indicating Cre-mediated recombination. Scale bar: 100 μm. [Figure 5] TEM image and table of hydrodynamic size, polydispersity index, and zeta potential of NT1-LNP. [Figure 6] This graph summarizes the relative fluorescence intensity of dissected brain tissue 1 hour after a single intravenous injection of 1 mg / kg DiR-labeled NT-LNPs. DiR was doped into NT-LNPs at a weight ratio of 10%. Mice were perfused with saline before dissection. One-way ANOVA, Sidak post-hoc analysis, *p<0.05 or **p<0.01. [Figure 7-1] Representative ex vivo fluorescence images of dissected brains 1 h after a single intravenous injection of 1 mg / kg DiR-labeled LNPs or NT1-O12B-doped NT LNPs (ratio 3:7, w / w) are shown, as well as the chemical structures of 76-O16B, EC16-80, and 113-O16B. DiR was doped into NT-LNPs at a 10% weight ratio. Mice were perfused with saline before dissection. [Figure 7-2] This is a continuation of Figure 7-1. [Figure 7-3] This is a continuation of Figure 7-2. [Figure 8-1] The chemical structures of NT-lipidoid and dimethyltryptamine are shown, as well as a representative ex vivo fluorescence image of a dissected brain 1 hour after a single intravenous injection of 1 mg / kg DiR-labeled NT-LNPs. DiR was doped into the NT-LNPs at a weight ratio of 10%. Mice were perfused with saline before dissection. [Figure 8-2] This is a continuation of Figure 8-1. [Figure 8-3] This is a continuation of Figure 8-2. [Figure 9]1 is a graph showing AmB concentrations in brain tissue 24 hours after intravenous injection of 5 mg / kg AmB in various NT1 derivatives, as measured using HPLC. Mice were perfused with saline before dissection. [Figure 10A] Photographs of AmB formulations in NT1-lipidoids with different tail lengths (O18B, O16B, O14B, O12B). All four NT1 / AmB inclusion bodies showed opaque suspensions. [Figure 10B] 1 is a graph showing the hydrodynamic diameter and polydispersity index of NT-LNPs determined by DLS measurements. [Figure 11] TEM images of the NT1-O12B / PBA-Q76O16B-3 / 7-AmB complexes, as well as a table summarizing the hydrodynamic size, polydispersity index, zeta potential, and DLC of the AmB / NT-LNP complexes. [Figure 12] This graph summarizes the relative fluorescence intensity of dissected brain tissue 1 hour after a single intravenous injection of 1 mg / kg DiR-loaded NT1-O12B / PBA-Q76-O16B LNPs. The weight ratio of DiR in the LNPs is 10%. **p<0.001. One-way ANOVA, Sidak post-hoc analysis. [Figure 13] Calibration curves of AmB concentrations dissolved in methanol ranging from 0.005 to 0.5 μg / mL (low concentrations) or 0.007 to 3.0 μg / mL (high concentrations) at 415 nm by HPLC. [Figure 14] 1 is a mAU-time graph of AmB concentration in brain tissue 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 by HPLC. [Figure 15-1] 1 is a graph showing AmB concentrations in other organs 24 hours after intravenous injection of 5 mg / kg AmB as measured by HPLC. [Figure 15-2] This is a continuation of Figure 15-1. [Figure 15-3] This is a continuation of Figure 15-2. [Figure 16]TEM images of blank and ASO-loaded NT1-O14B / 306-O12B-3 (ratio: 3 / 7) nanoparticles, and a table of hydrodynamic size, polydispersity index, and zeta potential. [Figure 17] TEM images of blank and (-27)GFP-Cre loaded NT1-O14B / PBAQ76O16B (ratio: 3 / 7) nanoparticles, and a table of hydrodynamic size, polydispersity index, and zeta potential. [Figure 18A] 1 is a scheme showing the synthesis of an IE tail. [Figure 18B] 1 is a scheme showing the synthesis of PBA-Q76O16B and PBA-Q80O16B. [Figure 18C] 1 is a scheme showing the synthesis of NT1-Neu. [Figure 19] Figures 19A-19N are fluorescent images of brain sections from Ail4 mice. Mice were injected with Cre mRNA complexed with Dlin-MC3 / NT1-O14B LNPs. The LNP formulation is described in slide 1. [Figure 20] Figures 20A-20B are fluorescent images of brain sections from Ail4 mice. Mice were injected with Cre mRNA complexed with PBA-Q76O16B / NT1-O14B LNPs. The LNP formulation is described in Slide 1. [Figure 21] 21A-21B are fluorescent images of brain sections from Ail4 mice. Mice were injected with Cre mRNA complexed with Dlin-MC3 / NT1-O14B LNPs. The LNP formulation is described in slide 1. DETAILED DESCRIPTION OF THE INVENTION

[0011] In one aspect, disclosed is a compound of formula I, or a pharmaceutically acceptable salt thereof: [ka] [In the formula, Y is a moiety derived from a neurotransmitter; W is -NR 20 -, -O-, or -S-; R 脂質 are 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 is heteroalkynyl; R 20 is R 脂質 , H., C. 1-6 Alkyl, C 1-6 Alkenyl, or C 1-6 alkynyl].

[0012] In certain embodiments, Y is [ka] is selected from.

[0013] In certain preferred embodiments, Y is [ka] is.

[0014] In certain embodiments, W is —NR 20 - or -S-. In certain embodiments, W is -NR 20 In certain embodiments, W is -S-.

[0015] In certain embodiments, W is —NR 20 - and R 20 is R 脂質 is.

[0016] In certain embodiments, W is —NR 20 - and R 20 is R 脂質 and Y is [ka] is

[0017] In certain embodiments, R 脂質 is of the following structure: [ka] [In the formula, R 1 and R 2 Each instance of is independently -H, -OH, -NHR 30 , or -SH; R 3 and R 4 are both -H; or R 3 and R 4 together form an oxo (=O) group; Z is -CH2-, -O-, or -NR 30 -, or -S-; X and Y are independently -CH2-, -NR 30 -, -O-, -S-, or -Se-; m is an integer selected from 1 to 3; n is an integer selected from 1 to 14; p is 0 or 1; q is an integer selected from 1 to 10; t is 0 or 1; R 30 -H, C 1-6 Alkyl, C 1-6 Alkenyl, or C 1-6 alkynyl].

[0018] In certain embodiments, R 1 and R 2 Each instance of is independently -H or -OH. In certain embodiments, R 1 and R 2 is —H. In certain embodiments, R 1 is -H; and R 2 is -OH.

[0019] In certain embodiments, R 3 and R 4is —H. In certain embodiments, R 3 and R 4 together form an oxo (=O) group.

[0020] In certain embodiments, Z is —CH 2 —, —O—, or —NR 30 In certain embodiments, Z is -CH2-. In certain embodiments, Z is -O-. In certain embodiments, Z is -NR 30 -It is.

[0021] In certain embodiments, R 1 and R 2 is -H and R 3 and R 4 together form an oxo (=O) group, and Z is O.

[0022] In certain embodiments, R 1 is -H and R 2 is -OH and R 3 and R 4 is -H and Z is -CH2-.

[0023] In certain embodiments, X and Y are independently -CH2- or -O-. In certain embodiments, X and Y are independently -CH2- or -O-, where X and Y are not the same. In certain embodiments, X and Y are independently -CH2- or -S-. In certain embodiments, X and Y are both -CH2-. In certain embodiments, X and Y are both -S-.

[0024] In certain embodiments, m is 1 or 2. In certain embodiments, m is 1. In certain embodiments, m is 2.

[0025] In certain embodiments, n is an integer selected from 4 to 12. In certain embodiments, n is an integer selected from 6 to 10.

[0026] In certain embodiments, p is 0. In certain embodiments, p is 1.

[0027] In certain embodiments, q is an integer selected from 2 to 8. In certain embodiments, q is an integer selected from 4 to 8.

[0028] In certain embodiments, t is 0. In certain embodiments, t is 1.

[0029] In certain embodiments, the compound is [ka] [ka] [ka] or a pharmaceutically acceptable salt thereof.

[0030] In certain aspects, lipidoid nanoparticles comprising the compounds described herein are disclosed.

[0031] In certain embodiments, the nanoparticles described herein further comprise a protein.

[0032] In a particular embodiment, the protein is GFP-Cre.

[0033] In certain embodiments, the nanoparticles described herein further comprise a nucleic acid.

[0034] In certain embodiments, the nucleic acid is a Tau-ASO.

[0035] In certain embodiments, the nanoparticles described herein further comprise a small molecule.

[0036] In some embodiments, the small molecule is an antifungal agent or a chemotherapeutic agent.

[0037] In certain embodiments, 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, vincristine sulfate, epirubicin, docetaxel, cyclophosphamide, carboplatin, levothyroxine, levothyroxine, levothyroxine hydrochloride ... Nalidomide, ibrutinib, abiraterone acetate, enzalutamide, pemetrexed, palbociclib, nilotinib, everolimus, ruxolitinib, epirubicin, pirirubicin, idarubicin, valrubicin, amrubicin, bleomycin, phleomycin, dactinomycin, mithramycin, streptozotecin, pentostatin, mitosins Selected from the group consisting of mitomycin C, enediynes, calicheamicin, glycosides, rebeccamycin, macrolide lactones, epothyphilone, ixabepilone, pentostatin, salinosporamide A, vinblastine, vincristine, etoposide, teniposide, vinorelbine, docetaxel, camptothecin, hycamtin, pederin, theopederin, annamide, trabectedin, aplidine, and ecteinascidin 743 (ET743).

[0038] In certain embodiments, the small molecule is amphotericin B or doxorubicin.

[0039] In certain embodiments, the lipidoid nanoparticles have a particle size of about 25 nm to about 1000 nm, hi certain embodiments, the lipidoid nanoparticles have a particle size of about 50 nm to about 500 nm.

[0040] In certain aspects, pharmaceutical compositions are disclosed that include the lipidoid nanoparticles described herein and a pharmaceutically acceptable carrier or excipient.

[0041] definition Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings commonly understood by those of ordinary skill in the art. Generally, the nomenclature used in connection with, and techniques of, chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics, and protein and nucleic acid chemistry described herein are those commonly used and well known in the art.

[0042] Unless otherwise indicated, the methods and techniques of the present disclosure are generally carried out according to the conventional methods well known in the art and described in various general and more specific references cited and discussed throughout this specification.See, for example, "Principles of Neural Science", McGraw-Hill Medical, New York, NY (2000); Motulsky, "Intuitive Biostatistics", Oxford University Press, Inc. (1995); Lodish et al., "Molecular Cell Biology, 4th ed.", WH Freeman & Co., New York (2000); Griffiths et al., "Introduction to Genetic Analysis, 7th ed.", WH Freeman & Co., NY (1999); and Gilbert et al., "Developmental Biology, 6th ed.", Sinauer Associates, Inc., Sunderland, MA (2000).

[0043] Chemical terms used herein, unless otherwise defined herein, are used in accordance with conventional usage in the art, as exemplified by "The McGraw-Hill Dictionary of Chemical Terms," ​​Parker S., Ed., McGraw-Hill, San Francisco, CA (1985).

[0044] As used herein, the term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes cases where the event or circumstance occurs as well as cases where it does not occur. For example, "optionally substituted alkyl" refers to cases where the alkyl may be substituted, and also refers to cases where the alkyl is not substituted.

[0045] It is understood that the substituents and substitution patterns on the compounds of the present invention can be selected by those skilled in the art to result in chemically stable compounds that can be easily synthesized from readily available starting materials using techniques known in the art and the methods described below. When a substituent is itself substituted with more than one group, it is understood that these multiple groups can be on the same carbon or on different carbons, as long as a stable structure is obtained.

[0046] As used herein, the term "optionally substituted" refers to the replacement of 1 to 6 hydrogen radicals in a given structure with the radical of a specified substituent, including, but not limited to: hydroxyl, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl, cycloalkyl, heterocyclyl, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH-O-alkyl, -OP(O)(O-alkyl), or -CH-OP(O)(O-alkyl). Preferably, "optionally substituted" refers to the replacement of 1 to 4 hydrogen radicals in a given structure with a substituent. More preferably, 1 to 3 hydrogen radicals are replaced by a substituent. It is understood that the substituent may be further substituted.

[0047] Articles such as "a," "an," and "the" can mean one or more unless indicated otherwise or clear from the context. A claim or description including "or" between one or more members of a group is considered to be satisfied when one, one or more, or all of the group members are present in, used in, or otherwise relevant to a given product or process, unless indicated to the contrary or clear from the context. The invention includes embodiments in which exactly one member of a group is present in, used in, or otherwise relevant to a given product or process. The invention includes embodiments in which one, more than one, or all of the group members are present in, used in, or otherwise relevant to a given product or process.

[0048] As used herein, the term "alkyl" refers to saturated aliphatic groups, including but not limited to, C1 to C6 10 Straight chain alkyl group or C1-C 10Branched alkyl groups are included. Preferably, the "alkyl" group refers to a C1-C6 straight chain alkyl group or a C1-C6 branched chain alkyl group. Most preferably, the "alkyl" group refers to a C1-C4 straight chain alkyl group or a C1-C4 branched chain 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 be substituted.

[0049] The term "acyl" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0050] The term "acylamino" is art-recognized and refers to an amino group substituted with an acyl group and may be represented, for example, by the formula hydrocarbylC(O)NH-.

[0051] The term "acyloxy" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0052] The term "alkoxy" refers to an alkyl group having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, and the like.

[0053] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and can be represented by the general formula alkyl-O-alkyl.

[0054] The term "alkyl" refers to saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. In preferred embodiments, a straight-chain or branched-chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C for a straight chain). 1-30 , C for branched chains 3-30 ), more preferably having 20 or fewer carbon atoms.

[0055] Furthermore, the term "alkyl," as used throughout the specification, examples, and claims, is intended to include both unsubstituted and substituted alkyl groups, the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone, and includes haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl.

[0056] "C x-y " or "C x ~C y The term "C" when used with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups containing x to y carbons in the chain. C alkyl indicates a hydrogen atom if the group is in a terminal position, or a bond if it is internal. 1-6 An alkyl group contains, for example, 1 to 6 carbon atoms in the chain.

[0057] The term "alkylamino," as used herein, refers to an amino group substituted with at least one alkyl group.

[0058] The term "alkylthio," as used herein, refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS-.

[0059] The term "amide" as used herein refers to the following group: [ka] In the formula, R9 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 complete a heterocycle having 4 to 8 atoms in the ring structure.

[0060] The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines and their salts, such as, for example: [ka] where R 9 , R 10 , and R 10 ' each independently represent a hydrogen or a hydrocarbyl group, or R 9 and R 10 together with the N atom to which they are attached complete a heterocycle having 4 to 8 atoms in the ring structure.

[0061] The term "aminoalkyl," as used herein, refers to an alkyl group substituted with an amino group.

[0062] The term "aralkyl," as used herein, refers to an alkyl group substituted with an aryl group.

[0063] As used herein, the term "aryl" includes substituted or unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon. Preferably, the ring is 5- to 7-membered, more preferably 6-membered. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings, where two or more carbons are common to two adjacent rings and at least one of the rings is aromatic, e.g., the other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

[0064] The term "carbamate" is art-recognized and refers to the following group: [ka] In the formula, R 9 and R 10 each independently represents hydrogen or a hydrocarbyl group.

[0065] The term "carbocyclylalkyl," as used herein, refers to an alkyl group substituted with a carbocyclic group.

[0066] The term "carbocycle" includes 5- to 7-membered monocyclic rings and 8- to 12-membered bicyclic rings. Each ring in a bicyclic carbocycle can be selected from saturated, unsaturated, and aromatic rings. Carbocycles include bicyclic molecules in which one, two, or more atoms are shared between the two rings. The term "fused carbocycle" refers to a bicyclic carbocycle in which each ring shares two adjacent atoms with the other ring. Each ring in a fused carbocycle can be selected from saturated, unsaturated, and aromatic rings. In an exemplary embodiment, an aromatic ring, such as 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, valence permitting, is included in the definition of carbocycle. Exemplary "carbocycles" 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 carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hept-3-ene. A "carbocycle" can be substituted at any one or more positions that can have a hydrogen atom.

[0067] The term "carbocyclylalkyl," as used herein, refers to an alkyl group substituted with a carbocyclic group.

[0068] The term "carbonate" is art-recognized and refers to the group --OCO.sub.2--.

[0069] The term "carboxy," as used herein, refers to a group represented by the formula -CO2H.

[0070] The term "ester" as used herein refers to the group -C(O)OR 9 where R 9 represents a hydrocarbyl group.

[0071] The term "ether" as used herein refers to a hydrocarbyl group bonded to another hydrocarbyl group via an oxygen atom. Thus, the ether substituent of a hydrocarbyl group can be hydrocarbyl-O-. Ethers can be symmetrical or asymmetrical. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include "alkoxyalkyl" groups, which can be represented by the general formula alkyl-O-alkyl.

[0072] The terms "halo" and "halogen," as used herein, mean halogen and include chloro, fluoro, bromo, and iodo.

[0073] The terms "hetaralkyl" and "heteroaralkyl," as used herein, refer to an alkyl group substituted with a heteroaryl group.

[0074] The terms "heteroaryl" and "hetaryl" include substituted or unsubstituted aromatic monocyclic ring structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, whose ring structures contain at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more cyclic rings, where two or more carbons are common to two adjacent rings and at least one of the rings is heteroaromatic; for example, the other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, and the like.

[0075] The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0076] The term "heterocyclylalkyl," as used herein, refers to an alkyl group substituted with a heterocycle group.

[0077] The terms "heterocyclyl," "heterocycle," and "heterocyclic" refer to a substituted or unsubstituted non-aromatic ring system, preferably a 3- to 10-membered ring, more preferably a 3- to 7-membered ring, whose ring system contains at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more cyclic rings, where two or more carbons are common to two adjacent rings and at least one of the rings is heterocyclic, e.g., the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

[0078] The term "hydrocarbyl," as used herein, refers to a group bonded through a carbon atom that has no =0 or =S substituents, typically having at least one carbon-hydrogen bond and a primarily carbon backbone, but may optionally contain heteroatoms. Thus, groups such as methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered hydrocarbyl for purposes of this application, while substituents such as acetyl (which has an =0 substituent on the linking carbon) and ethoxy (which is linked through an oxygen rather than a carbon) are not. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocyclic, heterocyclic, alkyl, alkenyl, alkynyl, and combinations thereof.

[0079] The term "hydroxyalkyl," as used herein, refers to an alkyl group substituted with a hydroxy group.

[0080] The term "lower," when used in conjunction with chemical moieties such as, for example, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups in which there are 10 or fewer, preferably 6 or fewer atoms in the substituent. "Lower alkyl," for example, refers to alkyl groups containing 10 or fewer, preferably 6 or fewer, carbon atoms. In certain embodiments, 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 descriptions of hydroxyalkyl and aralkyl (in which, for example, when counting carbon atoms in an alkyl substituent, atoms in the aryl group are not counted).

[0081] The terms "polycyclyl," "polycycle," and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl) in which two or more atoms are common to two adjacent rings, e.g., the rings are "fused rings." Each of the rings of a polycycle can be substituted or unsubstituted. In certain embodiments, each ring of a polycycle contains 3 to 10, preferably 5 to 7, atoms in the ring.

[0082] The term "sulfate" is art-recognized and refers to the group -OSO3H or a pharmaceutically acceptable salt thereof.

[0083] The term "sulfonamide" is art-recognized and may be represented by the general formula: [ka] wherein R 9 and R 10 independently represent hydrogen or hydrocarbyl.

[0084] The term "sulfoxide" is art-recognized and refers to the group --S(O)--.

[0085] The term "sulfonate" is art-recognized and refers to the group SO3H or a pharmaceutically acceptable salt thereof.

[0086] The term "sulfone" is art-recognized and refers to the group -S(O)2-.

[0087] The term "substituted" refers to moieties having substituents replacing a hydrogen on one or more backbone carbons. It will be understood that "substituted" or "substituted with" includes the implicit proviso that such substitution is subject to the permissible valencies of the substituted atom and substituent, and also results in a stable compound that does not spontaneously undergo transformation, for example, by rearrangement, cyclization, elimination, and the like. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valence of the heteroatom. Substituents can include any of the substituents described herein, for example, halogen, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate.

[0088] The term "thioalkyl," as used herein, refers to an alkyl group substituted with a thiol group.

[0089] The term "thioester" as used herein refers to the group -C(O)SR 9 or -SC(O)R 9 (In the formula, R 9 represents hydrocarbyl).

[0090] As used herein, the term "thioether" is equivalent to an ether where the oxygen has been replaced with a sulfur.

[0091] The term "urea" is art-recognized and has the general formula: [ka] where R 9 and R 10 independently represent hydrogen or hydrocarbyl.

[0092] 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.

[0093] The phrase "pharmaceutically acceptable" is art-recognized. In certain embodiments, this term includes compositions, excipients, adjuvants, polymers, and other materials and / or dosage forms that 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.

[0094] "Salt" is used herein to refer to an acid addition salt or a base addition salt.

[0095] Many of the compounds useful in the methods and compositions of the present disclosure have at least one stereocenter in their structure.This stereocenter can be in R or S configuration, and the R and S designations are used according to the rules described in Pure Appl. Chem. (1976), 45, 11-30.The present disclosure contemplates all stereoisomeric forms, such as enantiomeric and diastereomeric forms of compounds, salts, prodrugs or mixtures thereof (including mixtures of all possible stereoisomers).See, for example, International Publication No. 01 / 062726.

[0096] Furthermore, certain compounds containing alkenyl groups can exist as Z (Zusammen) or E (Entgegen) isomers. In each case, the present disclosure includes both mixtures and the separate individual isomers.

[0097] Some of the compounds may also exist in tautomeric forms, and such forms, although not explicitly shown in the formulas set forth herein, are intended to be included within the scope of the present disclosure.

[0098] "Pharmaceutically acceptable" means approved or approvable by a regulatory agency of the Federal or State government or its counterpart in a country other than the United States, or listed in the United States Pharmacopeia or other generally recognized pharmacopoeia for use in animals, especially humans.

[0099] "Pharmaceutically acceptable salts" refers to salts of the compounds of the present invention that are pharmaceutically acceptable and possess the desired pharmacological activity of the parent compound. In particular, such salts are non-toxic and can be inorganic or organic acid addition salts and base addition salts. Specifically, such salts include (1) salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or salts formed with 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, camphorsulfonic acid, 4-methyl Included are acid addition salts formed with organic acids such as bicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, t-butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, and the like; or (2) salts formed when an acidic proton present in the parent compound is replaced by a metal ion, such as an alkali metal ion, alkaline earth ion, or aluminum ion; or when coordinated with an organic base such as ethanolamine, diethanolamine, triethanolamine, N-methylglucamine, and the like. Salts further include, by way of example only, salts of non-toxic organic or inorganic acids such as sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and, if the compound contains a basic functional group, salts of hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate, and the like.

[0100] The term "pharmaceutically acceptable cation" refers to an acceptable cationic counterion of an acidic functional group. Examples of such cations include sodium, potassium, calcium, magnesium, ammonium, and tetraalkylammonium cations (see, e.g., Berge, et al., J. Pharm. Sci. 66 (1):1-79 (January 77)).

[0101] "Pharmaceutically acceptable vehicle" refers to a diluent, adjuvant, excipient, or carrier with which a compound of the invention is administered.

[0102] A "pharmaceutically acceptable metabolically cleavable group" refers to a group that is cleaved in vivo to yield a parent molecule of the structural formula shown herein. Examples of metabolically cleavable groups include -COR, -COOR, -CONRR, and -CHOR radicals, where R is independently selected in each occurrence from alkyl, trialkylsilyl, carbocyclic aryl, or carbocyclic aryl substituted with one or more of alkyl, halogen, hydroxy, or alkoxy. Specific examples of representative metabolically cleavable groups include acetyl, methoxycarbonyl, benzoyl, methoxymethyl, and trimethylsilyl groups.

[0103] "Prodrug" refers to a compound containing a cleavable group that becomes a pharmaceutically active compound of the present invention in vivo by solvolysis or under physiological conditions, including derivatives of the compounds of the present invention. Examples include, but are not limited to, choline ester derivatives, N-alkylmorpholine esters, and the like. Other derivatives of the compounds of the present invention are active in both their acid and acid derivative forms, but the acid-sensitive forms often offer advantages of solubility, tissue compatibility, or delayed release in mammalian organisms (see Bundgard, H., Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs include acid derivatives well known to those skilled in the art, such as esters prepared by reacting the parent acid with an appropriate alcohol, amides prepared by reacting the parent acid with a substituted or unsubstituted amine, acid anhydrides, or mixed anhydrides. Simple aliphatic or aromatic esters, amides, and anhydrides derived from acidic groups pendant on the compounds of the present invention are specific prodrugs. In some cases, it may be desirable to prepare double ester prodrugs, such as (acyloxy)alkyl esters or (alkoxycarbonyl)oxy)alkyl esters. In particular, the compounds of the present invention may be prodrugs of C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, aryl, C7-C 12 Substituted aryl and C7-C 12 It is an aryl alkyl ester.

[0104] "Solvate" refers to a form of a compound associated with a solvent or water (also called a "hydrate"), usually by a solvolysis reaction. This physical association also includes hydrogen bonding. Conventional solvents include water, ethanol, acetic acid, and the like. The compounds of the present invention may be prepared, for example, in crystalline form, and solvated or hydrated. Suitable solvates include pharmaceutically acceptable solvates, such as hydrates, and further include both stoichiometric and non-stoichiometric solvates. In certain instances, a "solvate" will be isolable, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. "Solvate" encompasses both solution-phase and isolable solvates. Representative solvates include hydrates, ethanolates, methanolates, and the like.

[0105] "Subjects" to which administration is contemplated include, but are not limited to, humans (male or female of any age, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, elderly adults)), and / or non-human animals, e.g., mammals such as primates (e.g., cynomolgus monkeys, rhesus monkeys), cows, pigs, horses, sheep, goats, rodents, cats, and / or dogs. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human animal. The terms "human," "patient," and "subject" are used interchangeably herein.

[0106] An "effective amount" refers to 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" may 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 for therapeutic treatment. Also, a "prophylactically effective amount" refers to an amount effective for prophylactic treatment.

[0107] "Preventing" or "prevention" or "prophylactic treatment" refers to reducing the risk of acquiring or developing a disease or disorder (i.e., preventing the development of at least one clinical symptom of the disease in a subject who has not yet been exposed to a disease-causing agent or is susceptible to the disease prior to the onset of the disease).

[0108] The term "prophylaxis" is related to "prevention" and refers to measures or treatments aimed at preventing, rather than treating or curing, a disease. Non-limiting examples of preventive measures can include the administration of a vaccine; the administration of low-molecular-weight heparin to hospitalized patients who are at risk of thrombosis due to immobility; and the administration of antimalarial drugs such as chloroquine prior to visiting areas where malaria is endemic or where there is a high risk of contracting malaria.

[0109] "Treating" any disease or disorder or "treatment" or "therapeutic treatment" of a disease or disorder refers, in one embodiment, to ameliorating the disease or disorder (i.e., halting the disease or reducing at least one sign, degree, or severity of its clinical symptoms). In another embodiment, "treating" or "treatment" refers to improving at least one physical parameter, which may not be discernible by the subject. In yet another embodiment, "treating" or "treatment" refers to modulating the disease or disorder physically (e.g., stabilization of discernible symptoms), physiologically (e.g., stabilization of physical parameters), or both. In a further embodiment, "treating" or "treatment" relates to slowing the progression of the disease.

[0110] As used herein, the term "isotopic variant" refers to a compound that contains unnatural proportions of isotopes at one or more of the atoms that constitute such compound. For example, an "isotopic variant" of a compound may contain, for example, deuterium ( 2 H or D), carbon 13 ( 13 C), nitrogen 15(15 In compounds with such isotopic substitutions, if present, the following atoms may be present: 2 H / D, and any carbon 13 C, or any nitrogen 15 It will be understood that the presence and location of such atoms may vary, such as N, and that the determination of such atoms is within the skill of one of ordinary skill in the art. Similarly, the present invention includes the preparation of isotopic variants using radioactive isotopes, for example, when the resulting compounds may be used in drug and / or substrate tissue distribution studies. The radioactive isotope tritium ( 3 H) and carbon-14 ( 14 C) is particularly useful for this purpose in view of its ease of incorporation and ease of detection. 11 C. 18 F, 15 O. 13 Compounds substituted with positron emitting isotopes, such as N, can also be prepared and are useful 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 invention.

[0111] It should also be understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed "isomers." Isomers that differ in the arrangement of their atoms in space are termed "stereoisomers."

[0112] Stereoisomers that are not mirror images of one another are called "diastereomers," while those that are non-superimposable mirror images of each other are called "enantiomers." When a compound has an asymmetric center, for example, if it is bonded to four different groups, a pair of enantiomers is possible. Enantiomers are characterized by the absolute configuration of their asymmetric center and described by the R- and S-sequencing rules of Cahn and Prelog or by the way the molecule rotates the plane of polarized light, designated as dextrorotatory or levorotatory (i.e., (+)- or (-)-isomer, respectively). Chiral compounds can exist as individual enantiomers or as mixtures thereof. A mixture containing equal proportions of enantiomers is called a "racemic mixture."

[0113] "Tautomers" are interchangeable forms of a particular compound structure, differing in the displacement of hydrogen atoms and electrons. Thus, two structures can be in equilibrium through the movement of electrons and atoms (usually H). For example, enols and ketones are tautomers because they are rapidly interconverted by treatment with either acid or base. Another example of tautomerism is the aci- and nitro-forms of phenylnitromethane, which are similarly formed by treatment with acid or base. Tautomers can be relevant to achieving optimal chemical reactivity and biological activity of a compound of interest.

[0114] As used herein, a pure enantiomer compound is substantially free of other enantiomers or stereoisomers of the compound (i.e., in enantiomeric excess). In other words, the "S" form of a compound is substantially free of the "R" form of the compound and is thus in enantiomeric excess of the "R" form. The terms "enantiomerically pure" or "pure enantiomer" mean that a compound contains greater than 95%, 96%, 97%, 98%, 98.5%, 99%, 99.2%, 99.5%, 99.6%, 99.7%, 99.8%, or 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.

[0115] As used herein, unless otherwise indicated, the term "enantiomerically pure R compounds" refers to at least about 95% by weight R compounds and at most about 5% by weight S compounds, at least about 99% by weight R compounds and at most about 1% by weight S compounds, or at least about 99.9% by weight R compounds and at most about 0.1% by weight S compounds. In certain embodiments, the weights are based on the total weight of the compounds.

[0116] As used herein, unless otherwise indicated, the term "enantiomerically pure S-compounds" or "S-compounds" refers to at least about 95% by weight S-compounds and at most about 5% by weight R-compounds, at least about 99% by weight S-compounds and at most about 1% by weight R-compounds, or at least about 99.9% by weight S-compounds and at most about 0.1% by weight R-compounds. In certain embodiments, the weights are based on the total weight of the compounds.

[0117] As used herein, an enantiomerically pure compound, or its pharmaceutically acceptable salt, solvate, hydrate, or prodrug, may be present together with other active or inactive ingredients. For example, a pharmaceutical composition containing an enantiomerically pure R compound may contain, for example, about 90% excipients and about 10% enantiomerically pure R compound. In some embodiments, the enantiomerically pure R compound in such a composition may contain, for example, at least about 95% by weight of the R compound and up to about 5% by weight of the S compound, based on the total weight of the compound. For example, a pharmaceutical composition containing an enantiomerically pure S-compound may contain, for example, about 90% excipients and about 10% of the enantiomerically pure S compound. In some embodiments, the enantiomerically pure S compound in such a composition may contain, for example, at least about 95% by weight of the S compound and up to about 5% by weight of the R compound, based on the total weight of the compound. In some embodiments, the active ingredient may be formulated with little or no excipients or carriers.

[0118] The compounds of the present invention may possess one or more asymmetric centers; therefore, such compounds can be produced as individual (R)- or (S)-stereoisomers or as mixtures thereof.

[0119] Unless otherwise indicated, the description or naming of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures, racemic or otherwise, of such compounds. The methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art.

[0120] Those skilled in the art of organic synthesis will recognize that the maximum number of heteroatoms in a stable, chemically feasible heterocyclic ring, whether aromatic or non-aromatic, is determined by the ring size, degree of unsaturation, and valence of the heteroatoms. Generally, a heterocyclic ring can have from 1 to 4 heteroatoms, so long as the heteroaromatic ring is chemically feasible and stable. [Example]

[0121] In order that the invention described herein may be more fully understood, the following examples are set forth. The examples described herein are provided to illustrate the compounds, compositions, materials, devices, and methods provided herein, and should not be construed in any way as limiting the scope thereof.

[0122] material and method general : All chemicals used for lipid synthesis were purchased from Sigma-Aldrich and used directly. All ASOs and DNA fragments were purchased from Integrated DNA Technologies (IDT). ASOs were provided by IDT, and where noted, ASO products provided by the company were used to include chemical modifications to improve stability. HeLa-DsRed and GFP-HEK cells were maintained in Dulbecco's modified Eagle's 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 using a flow cytometer (BD FACS Calibur, BD Science, CA). The (-27)GFP-Cre (addgene #89253) protein was expressed and extracted from BL21 Escherichia coli (E. coli) and further purified using a Ni-NTA column (Qiagen). Nanoparticle size and zeta potential were recorded using a ZetaPALS particle size analyzer. TEM images were taken using a Technai Spirit transmission electron microscope (FEI).

[0123] lipid synthesis All head amines used in lipid synthesis were commercially available from Sigma-Aldrich. All cationic lipidoids (NT1-O12B-O18B, NT2-O12B-O18B, NT3-O12B-O18B, NT1-EC16, NT1-C18, NT1-IE, NT2-EC16, NT2-IE, NT3-EC16, NT3-IE, 306-O12B-3, 76-O16B) were synthesized according to our previous report. The crude products were purified using flash chromatography on silica gel. The 1E tail was synthesized as shown in Figure 18A. The phenylboronic acid quaternized lipidoid was synthesized as shown in Figure 18B. NT1-Neu was synthesized as shown in Figure 18C. 1H NMR and electrospray ionization (ESI) MS were used to confirm the lipid structures.

[0124] Biodistribution of DiR-labeled NT-LNPs in the mouse brain NT-lipidoid and DiR were dissolved together in 100% ethanol at a weight ratio of 10:1. Then, 100 μL of the solution was added dropwise to 300 μL of sodium acetate buffer (25 mM, pH 5.2) and briefly vortexed. Finally, the ethanol in the formulation was removed by dialysis (MWCO 35 kDa, ThermoFisher) against diH2O (deionized water) for 12 hours. The DiR-labeled LNP was then intravenously injected into BALB / C mice (female, 6 weeks old). After 1 hour, the mice were anesthetized and perfused with saline. The mouse brains were then harvested. The fluorescent signal distribution was visualized using a Spectrum CT Biophotonic Imager (PerkinElmer, Boston, MA).

[0125] Preparation of AmB / NT-lipidoid nanoparticle formulations AmB was prepared according to our previous report. 3Briefly, 1 mg of each lipidoid (solid) was mixed with 1 mg of AmB in 300 μL of dimethyl sulfoxide (DMSO). The mixture was sonicated for 30 min and then vortexed for 10 min until completely dissolved. The solution was added dropwise to a glass bottle containing 600 μL of sodium acetate buffer (pH 5.0) with continuous homogenization at 700 rpm. The solution was then dialyzed overnight against distilled water using dialysis tubing (MWCO 35 kDa) to remove DMSO and unencapsulated AmB.

[0126] Characterization of AmB / NT-lipidoid nanoparticulate formulations The particle size and polydispersity index (PDI) of all inclusion bodies were measured using dynamic light scattering (DLS). Zeta potential was recorded using a ZetaPALS particle size distribution analyzer. DLC of AmB was calculated according to our previous report. 3 TEM images were taken with a Technai Spirit transmission electron microscope from FEI.

[0127] statistical analysis Statistical analysis was performed using one-way analysis of variance (ANOVA) followed by the Turkey-Kramer multiple comparison test for more than two groups. Student's t-test was used to compare two groups using Prism (v.8, GraphPad Software, La Jolla, CA). A value of p<0.05 was considered significant.

[0128] Synthesis of NT-lipidoid and BBB permeability study of NT-LNP The neurotransmitters tryptamine, phenethylamine, and phenylethanolamine were selected as the structural basis for the synthetic lipidoids. NT-lipidoids were synthesized by Michael addition between the primary amines of neurotransmitters and acrylate-containing hydrophobic tails in glass vials at 70 °C for 48 h, using an approach similar to our previously published combinatorial lipid library synthesis strategy (Figure 1B). The result is a combinatorial library of NT-lipidoids, each containing one specific neurotransmitter as the head group and one specific bioreducible hydrophobic structure as the tail group. NT-lipidoids are designated "NTn-O[x]B" (n = 1, 2, 3), where NT1 is tryptamine, NT2 is phenethylamine, NT3 is phenylethanolamine, and O[x]B represents the bioreducible hydrophobic tail, where [x] indicates the number of carbon atoms in the acrylate hydrophobic tail shown in Figure 1B. For example, NT1-O12B represents a lipidoid containing a tryptamine head group and a hydrophobic tail group containing 12 carbon atoms. All NT-lipidoids were purified using flash chromatography and characterized by ESI-MS (Figure 5). The resulting NT-lipidoids are amphiphilic and therefore can self-assemble into either micelles or liposomes when prepared in aqueous solution. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) of the NT-lipidoids showed that these structures indeed self-assembled into spherical liposome-like structures (Figure 6).

[0129] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0130] Using a fluorescent dye (DiR) as a model cargo, we further investigated whether these NT-lipidoids could cross the BBB during systemic intravenous delivery. Hydrophobic small molecules, such as DiR, can partition into the hydrophobic regions of micelles and liposomes and are often used to track the biodistribution of these structures. To formulate DiR-loaded NT-lipidoid, NT-lipidoid and DiR were mixed in ethanol at a 10 / 1 (w / w) ratio, the mixture was added dropwise to sodium acetate buffer (25 mM, pH 5.2), and the ethanol was then removed by dialysis. The DiR-loaded NT-lipidoid nanoparticle solution was injected into mice via tail vein injection. After 1 h, the animals were sacrificed 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.

[0131] As shown in Figure 1C, a strong DiR fluorescence signal is observed in the brains of mice treated with DiR / NT1-lipidoid nanoparticles, compared to the brains of mice treated with DiR / NT2-lipidoid and DiR / NT3-lipidoid, which show very weak fluorescence signals. NT1-lipidoids containing shorter aliphatic chain lengths yield greater fluorescence intensity, and it is also observed that the length of the aliphatic tail chain has a significant impact on the observed fluorescence intensity (Figure 7). There are no significant differences in physical properties, such as hydrodynamic size, polydispersity index, zeta potential, and morphology, between these NT1-derived lipidoids (Figure 6).

[0132] We hypothesized that doping NT1-lipidoids, such as NT1-O12B, into other BBB-impermeable lipid formulations would allow the resulting lipid formulations to cross the BBB. To examine their ability to deliver DiR to the brain, we used previously described synthetic lipids, 76-O16B, EC16-80, and 113-O16B. We found that none of these lipids alone was effective in delivering DiR to the mouse brain, but when these lipids were doped with NT1-O12B, strong DiR signals were observed in the mouse brain (Figure 8).

[0133] 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 membrane. 21 Our results are also driven by active transport, suggesting that changes in the chemical structure of the NT1 lipid may modulate its ability to cross the BBB. We specifically hypothesized that the ionizability of the α-amine of tryptamine (NT1) after lipidation is a key factor for the derivative's ability to cross the BBB. To test this hypothesis, we synthesized a series of NT1 derivatives with different linkers, as shown in Figure 9. DiR signals were observed in the brains of mice treated with all NT1 derivatives except NT1-neu. In NT1-neu, the α-amine of tryptamine is linked via an amide bond and is not ionizable, whereas the α-amines of all other NT1 derivatives are ionizable. Furthermore, no strong DiR signals were observed in the brains of mice treated with NT2- and NT3-derived lipidoids with any linker.

[0134] Delivery of the small molecule AmB to the mouse brain As shown above, NT1-derived lipidoids were identified to be capable of delivering a hydrophobic dye (DiR) to the brain, either when used alone or when doped into other LNPs. These NT1-derived lipidoids were used to investigate the brain delivery of therapeutically relevant hydrophobic drug molecules. Amphotericin B (AmB) was selected as a model drug. AmB is a classic polyene antifungal drug and is the gold standard for the treatment of severe systemic fungal infections. However, AmB cannot be used clinically for the treatment of cerebral fungal infections due to its BBB impermeability. Recently, we formulated AmB into synthetic lipidoid nanoparticles and performed thorough PK and biodistribution studies of the AmB formulation using conventional synthetic lipid nanoparticles. However, in that study, none of our lipid nanoparticles were able to penetrate the BBB and deliver AmB to the mouse brain. 27 .

[0135] Using a procedure similar to DiR encapsulation, AmB was encapsulated in pure NT1-lipidoids (i.e., NT1-O12B, NT1-O14B, NT1-O16B, and NT1-O18B). AmB-loaded NT1-lipidoid nanoparticles were injected into mice via the tail vein at a dose of 5 mg / kg AmB per mouse. After 24 h, the animals were sacrificed, and their brains were harvested, perfused with saline, and homogenized. The concentration of AmB in brain tissue was quantified using HPLC (detailed methods are in the SI). As shown in Figure 10, the AmB concentration in brain tissue from all four groups was approximately 150 ng / g tissue. Notably, our previous report showed that AmB was undetectable in the brain after systemic delivery with traditional synthetic lipidoids, indicating that the NT1-lipidoid formulation enhanced AmB delivery to the mouse brain.

[0136] However, AmB formulated in NT1-lipidoid formulations showed an opaque solution (Figure 11A), indicating large particle sizes in the solution. DLS results (Figure 11B, Figure 12) showed that the nanoparticles ranged in diameter from 750 to 800 nm. It is speculated that making NT1-lipidoid nanoparticles smaller may help improve brain delivery efficiency. Previous reports have found that quaternized lipidoids provided stable AmB formulations with smaller particle sizes compared to non-quaternized lipids. 27 Therefore, it is hypothesized that doping quaternized lipidoids with NT1-lipidoids may result in smaller nanoparticle sizes while maintaining or improving their ability to penetrate the BBB.

[0137] Here, we synthesized a new phenylboronic acid quaternized lipidoid, PBA-Q76-O16B (Figure 2A), for AmB encapsulation. NT1-O12B exhibited the highest DiR fluorescence intensity among all NT lipidoids (Figure 1C), and therefore was selected as a dopant for enhanced brain delivery. AmB was formulated in a mixture of NT1-O12B and PBA-Q76-O16B, in which the two lipidoids were mixed at different weight ratios (7:3, 5:5, 3:7, 1:9, and pure PBA-Q76-O16B). As shown in Figure 2B, AmB inclusion bodies gradually became a homogeneous, transparent, yellow solution as the proportion of PBA-Q76-O16B lipidoid in the formulation increased. The hydrodynamic size also decreased from 800 nm to 100 nm (Figure 2C, Figure 12). Using DiR as the cargo, we observed that a lipidoid containing NT1-O12B and PBA-Q76-O16B at a 3:7 (w / w) ratio produced the strongest fluorescent signal in mouse brains compared to all other lipid ratios (Figure 2D). The fluorescent signal intensity at the 3:7 ratio was 4.5-fold higher than that of brains treated with DiR formulated in pure NT1-O12B (Figure 13). We further investigated AmB delivery using mixed lipids and determined the AmB concentration in mouse brain tissue 24 hours after intravenous injection of 5 mg / kg AmB per mouse. As shown in Figure 2E, the amount of AmB detected in the brain increased as the doping ratio of PBA-Q76-O16B increased from 0% (i.e., pure NT1-O12B) to 70% (i.e., a 3:7 ratio), reaching a maximum concentration of approximately 300 ng / g, which was approximately two-fold higher than that of pure NT1-O12B. When the doping ratio was further increased to 90% (i.e., 1:9), the AmB concentration decreased slightly but was still higher than that of AmB formulated in pure NT1-O12B. Thus, the AmB delivery results were in excellent agreement with those of DiR delivery (Figures 2D and 2E). Interestingly, without doping with NT1-lipidoid, AmB was barely detectable in the brain after intravenous injection of pure PBA-Q76-O16B / AmB.These results demonstrated the important role of NT1 lipidoids in facilitating brain delivery and the importance of finding the optimal doping ratio.

[0138] Delivery of Tau ASOs to the mouse brain for gene knockdown We evaluated the efficiency of mixed lipidoid formulations for ASO delivery in vitro by delivering an ASO targeting green fluorescent protein (GFP) mRNA into HEK cells stably expressing GFP (Figure 3B). NT1-O14B alone did not exhibit GFP silencing effects (10:0 ratio in Figure 3B), indicating that this lipidoid alone is ineffective at delivering ASOs into cells. However, GFP silencing was observed when ASOs were delivered using LNPs containing a mixture of NT1-O14B and 306-O12B-3. When the doping ratio of 306-O12B-3 was greater than 50% (i.e., a weight ratio of 5:5 or greater favoring 306-O12B-3), GFP silencing in GFP-HEK cells was observed, and silencing efficiency increased as the 306-O12B-3 doping ratio increased. A scrambled ASO delivered by lipofetamine 2000 (LPF 2K) did not exhibit GFP silencing, demonstrating that GFP silencing is truly ASO sequence-specific.

[0139] We then investigated whether a mixed lipidoid formulation (NT1-O14B and 306-O12B-3) could deliver ASOs to the brain and mediate gene knockdown in vivo. Because ASO-mediated tau reduction after local injection of tau-ASOs using an intracerebroventricular (ICV) pump has shown promising results in the treatment of Alzheimer's disease (AD), we selected tau as a therapeutic target and designed an ASO targeting tau mRNA. 31、32 .

[0140] The sequence of the tau-ASO was obtained from published literature. 31The tau ASOs were selected according to the methodology and synthesized by IDT. To improve efficacy, we provided tau ASOs containing phosphorothioate groups between each nucleotide and 2'-O-methoxyethyl modification of the riboses in the 5' and 3' terminal nucleotides. To formulate ASOs for intravenous injection, the ASOs were mixed with the formulated LNP solution at a weight ratio of 1 / 15 (ASO to total lipid). Each mouse received five injections of 1 mg / kg ASO, with each injection spaced 3 days apart. Four days after the last injection, the mice were sacrificed and perfused. Brain tissue was harvested, homogenized, and total RNA was extracted. Total tau mRNA levels were analyzed by quantitative PCR. As shown in Figure 3C, no reduction in tau mRNA was detected in brain tissue when ASOs were delivered using either pure NT1-O14B or pure 306-O12B-3. For mixed lipidoid formulations, only the 5:5 and 3:7 w / w ratios of NT1-O14B and 306-012B-3 demonstrated a reduction in tau mRNA in the brain. These two formulations resulted in approximately 25% and 50% mRNA reduction, respectively. No tau mRNA silencing was observed with other ratios (i.e., 7:3 and 1:9) of mixed lipidoid formulations.

[0141] To confirm that ASO delivery resulted in functional tau knockdown, we also examined tau protein levels in ASO-treated mice using ELISA (Figure 3D). Compared with the untreated group, mice treated with tau-ASO formulated in NT1-O14B / 306-O12B-3 (3:7 w / w) showed substantially reduced total tau protein levels. Furthermore, using the exact same method as for functional ASO, we delivered scrambled tau-ASO at the best ratio (NT1-O14B / 306-O12B-3, 3:7 w / w). As shown, neither tau mRNA silencing effect nor tau protein reduction was detected, demonstrating that tau knockdown was specifically due to sequence-specific ASO silencing.

[0142] Delivery of GFP-Cre fusion protein for gene recombination in the brain of Ai14 mice GFP-fused Cre recombinase was selected as a model protein for studies using the Ail4 mouse model line (Figure 4A). The Ail4 mouse line contains a flox-stop-flox tdTomato construct. Successful intracellular delivery of the Cre protein into Ail4 mouse cells results in genetic recombination, turning on tdTomato expression, which can be directly visualized as a red fluorescent signal without additional staining. Here, we used the (-27)GFP-Cre protein. NT1-O14B LNPs doped with PBA-Q76-O16B were selected because these nanoparticles were able to successfully deliver (-27)GFP-Cre. The weight ratio of NT1-O14B to PBA-Q76-O16B was fixed at 3:7 based on the results observed from AmB and ASO delivery. Lipid formulations were prepared using the approach described for formulations for ASO delivery. Briefly, (-27)GFP-Cre protein and LNP were mixed at a 1:4 weight ratio, and the solution was incubated at room temperature for 15 minutes before intravenous injection. Mice were injected four times at a dose of 50 μg of protein per injection. Five days after the final injection, mice were sacrificed, and brain tissue was harvested, fixed, and dehydrated. The tissue was then cryosectioned into 15 μm slices and counterstained with DAPI for fluorescent imaging. As shown in Figure 4B, strong tdTomato signals were observed in multiple regions of the brain, including the cerebral cortex, hippocampus, and cerebellum. In contrast, no tdTomato expression was observed in the brain of mice injected with LNP formulations using either pure NT1-O14B (10:0) or pure PBA-Q76-O16B (0:10).

[0143] Delivery of different formulations of GFP-Cre fusion protein for gene recombination in the brain of Ai14 mice NT1-O14B was doped into various lipid nanoparticle formulations, including 306-O12B, PBA-Q76O16B, and Dlin-MC3, and the doped LNP formulations were examined for Cre mRNA delivery to the brains of Ai14 mice via intravenous (i.v.) injection. The weight ratio of NT1-O14B to other ionizable lipids (e.g., 306-O12B, PBA-Q76O16B, and Dlin-MC3) was 3:7. To form stable LNPs, other co-lipids, including 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG2000), cholesterol, and DOPE, were also included. Cre recombinase-encoding mRNA was loaded into the LNPs and injected into Ai14 mice. Mice were sacrificed at specific time points, and brain tissue was collected, fixed, and dehydrated. The tissue was then cryosectioned into 15 μm slices and counterstained with DAPI for fluorescence imaging. tdTomato signals were observed in multiple brain regions, demonstrating successful delivery of Cre mRNA to brain cells using this LNP formulation via systemic injection. Fluorescence images of brain sections from Ai14 mice are shown in Figures 19A-19N, 20A-20B, and 21A-21B. NT1-O14B-doped 306-O12B showed the highest brain delivery compared to PBA-O76O16B or Dlin-MC3 LNPs. References

[0144] 1. Pardridge, WM Blood-brain barrier delivery. Drug Discov. Today 12, 54-61 (2007). 2. Barchet, TM & Amiji, MM Challenges and opportunities in CNS delivery of therapeutics for neurodegenerative diseases. Expert Opin. Drug Del. 6, 211-225 (2009). 3. Collins, P.Y. et al. Grand challenges in global mental health. Nature 475, 27-30 (2011). 4. Obermeier, B., Daneman, R. & Ransohoff, R.M. Development, maintenance and disruption of the blood-brain barrier. Nat. Med. 19, 1584-1596 (2013). 5. Khorkova, O. & Wahlestedt, C. Oligonucleotide therapies for disorders of the nervous system. Nat. Biotechnol. 35, 249-263 (2017). 6. Patel, M.M. & Patel, B.M. Crossing the blood-brain barrier: recent advances in drug delivery to the brain. CNS Drugs 31, 109-133 (2017). 7. Dong, X. Current strategies for brain drug delivery. Theranostics 8, 1481-1493 (2018). 8. Fung, L.K., Shin, M., Tyler, B., Brem, H. & Saltzman, W.M. Chemotherapeutic drugs released from polymers: distribution of 1, 3-bis (2-chloroethyl)-l-nitrosourea in the rat brain. Pharm. Res. 13, 671-682 (1996). 9. Rubenstein, J.L. et al. Phase I study of intraventricular administration of rituximab in patients with recurrent CNS and intraocular lymphoma. J. Clin. Oncol. 25, 1350-1356 (2007). 10. Lu, C.-T. et al. Current approaches to enhance CNS delivery of drugs across the brain barriers. Int. J. Nanomedicine 9, 2241-2257 (2014). 11. Villringer, K. et al. DCE-MRI blood-brain barrier assessment in acute ischemic stroke. Neurology 88, 433-440 (2017). 12. Fu, H. & McCarty, D.M. Crossing the blood-brain-barrier with viral vectors. Curr. Opin. Virol. 21, 87-92 (2016). 13. Ha, D., Yang, N. & Nadithe, V. Exosomes as therapeutic drug carriers and delivery vehicles across biological membranes: current perspectives and future challenges. Acta Pharm. Sin. B 6, 287-296 (2016). 14. Pardridge, W.M. Drug and gene targeting to the brain with molecular Trojan horses. Nat. Rev. Drug Discov. 1, 131-139 (2002). 15. Zhou, Y., Peng, Z., Seven, E.S. & Leblanc, R.M. Crossing the blood-brain barrier with nanoparticles. J. Control. Release 270, 290-303 (2018). 16. Mingozzi, F. & High, K.A. Immune responses to AAV vectors: overcoming barriers to successful gene therapy. Blood 122, 23-36 (2013). 17. Pardridge, W.M. Delivery of biologics across the blood-brain barrier with molecular Trojan horse technology. BioDrugs 31, 503-519 (2017). 18. Blanco, E., Shen, H. & Ferrari, M. Principles of nanoparticle design for overcoming biological barriers to drug delivery. Nat. Biotechnol. 33, 941-951 (2015). 19. Wen, J. et al. Sustained delivery and molecular targeting of a therapeutic monoclonal antibody to metastases in the central nervous system of mice. Nat. Biomed. Eng. 3, 706-716 (2019). 20. Snowman, A.M. & Snyder, S.H. Cetirizine: actions on neurotransmitter receptors. J. Allergy Clin. Immunol. 86, 1025-1028 (1990). 21. Carbonaro, T.M. & Gatch, M.B. Neuropharmacology of N, N-dimethyltryptamine. Brain Res. Bull. 126, 74-88 (2016). 22. Wang, M. et al. Efficient delivery of genome-editing proteins using bioreducible lipid nanoparticles. Proc. Natl. Acad. Sci. U.S.A. 113, 2868-2873 (2016). 23. Chang, J. et al. Integrating Combinatorial Lipid Nanoparticle and Chemically Modified Protein for Intracellular Delivery and Genome Editing. Acc. Chem. Res. 52, 665-675 (2018). 24. Wang, M., Alberti, K., Sun, S., Arellano, C.L. & Xu, Q. Combinatorially designed lipid-like nanoparticles for intracellular delivery of cytotoxic protein for cancer therapy. Angew. Chem. Int. Ed. 53, 2893-2898 (2014). 25. Ostrosky-Zeichner, L., Marr, K.A., Rex, J.H. & Cohen, S.H. Amphotericin B: time for a new" gold standard". Clin. Infect. Dis. 37, 415-425 (2003). 26. Xu, N. et al. Efficacy of intravenous amphotericin B-polybutylcyanoacrylate nanoparticles against cryptococcal meningitis in mice. Int. J. Nanomedicine 6, 905-913 (2011). 27. Liu, F. et al. In vitro and in vivo study of an Amphotericin B formulation with quaternized bioreducible lipidoids. ACS Biomater. Sci. Eng. 6, 1064-1073 (2020). 28. Rinaldi, C. & Wood, M.J. Antisense oligonucleotides: the next frontier for treatment of neurological disorders. Nat. Rev. Neurol. 14, 9-21 (2018). 29. Talbot, K. & Wood, M.J. Wrangling RNA: Antisense oligonucleotides for neurological disorders. Sci. Transl. Med. 11, eaay2069 (2019). 30. Yang, L. et al. Efficient delivery of antisense oligonucleotides using bioreducible lipid nanoparticles in vitro and in vivo. Mol. Ther. Nucleic Acids (2020). https: / / doi.org / 10.1016 / j.omtn.2020.01.018 31. DeVos, SL et al. Antisense reduction of tau in adult mice protects against seizures. J. Neurosci. 33, 12887-12897 (2013). 32. DeVos, SL et al. Tau reduction prevents neuronal loss and reverses pathological tau deposition and seeding in mice with tauopathy. Sci. Transl. Med. 9, eaag0481 (2017).

[0145] Incorporation by Reference All U.S. and PCT patent publications and U.S. patents referred to herein are incorporated by reference in their entirety, as if each individual patent publication or patent were specifically and individually indicated to be incorporated by reference. In the case of conflict, the present application, including any definitions herein, will control.

[0146] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments described herein. The scope of the embodiments described herein is not intended to be limited to the above description, but rather is as set forth in the appended claims. Those skilled in the art will appreciate that various changes and modifications to this description can be made without departing from the spirit or scope of the invention, as defined in the following claims. Finally, preferred embodiments of the present invention are described in sections. [Embodiment 1] A compound of formula I or a pharmaceutically acceptable salt thereof: [ka] [In the formula, Y is a moiety derived from a neurotransmitter; W is -NR 20 -, -O-, or -S-; R 脂質 are 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 is heteroalkynyl; R 20 is R 脂質 , H., C. 1-6 Alkyl, C 1-6 Alkenyl, or C 1-6 alkynyl]. [Embodiment 2] Y is

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Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof: 【Chemistry 1】 [In the formula, Y is, 【Chemistry 2】 Selected from: W is -NR ２０ - and; R 脂質 teeth, 【Transformation 3】 and Each instance of R 1 and R 2 is independently —H, —OH, —NHR 30 , or —SH; R 3 and R 4 are both —H; or R 3 and R 4 together form an oxo (═O) group; Z is —CH 2 —, —O—, —NR 30 —, or —S—; X and Y 1 are independently —CH 2 —, —NR 30 —, —O—, —S—, or —Se—; m is an integer selected from 1 to 3; n is an integer selected from 1 to 14; p is 0 or 1; q is an integer selected from 1 to 10; t is 0 or 1; R 30 is —H, C 1-6 alkyl, C 1-6 alkenyl, or C 1-6 alkynyl; R ２０ is R 脂質 and However, if Y is 【Chemistry 4】 where R 1 and R 2 are —H, R 3 and R 4 together form an oxo (═O) group, and Z is O; or R 1 is —H, R 2 is —OH, R 3 and R 4 are —H, and Z is —CH 2 —.

2. Y is 【Transformation 5】 2. The compound of claim 1, wherein:

3. Y is 【Transformation 6】 2. The compound of claim 1, wherein:

4. Y is 【Transformation 7】 2. The compound of claim 1, wherein:

5. R １ and R ２ 4. The compound of claim 2 or 3, wherein each instance of is independently -H or -OH.

6. R １ and R ２ The compound of claim 2 or 3, wherein is —H.

7. The compound of any one of claims 2, 3, 5, and 6, wherein R 3 and R 4 are —H.

8. The compound of any one of claims 2, 3, 5, and 6, wherein R 3 and R 4 together form an oxo (═O) group.

9. Z is -CH ２ -, -O-, or -NR ３０ The compound according to any one of claims 2, 3, and 5 to 8, wherein 10. The compound of any one of claims 2, 3, and 5 to 8, wherein Z is -CH 2 -.

11. The compound of any one of claims 2, 3, and 5-8, wherein Z is -O-.

12. The compound of any one of claims 2, 3, and 5 to 8, wherein Z is -NR 30 -.

13. R １ and R ２ is -H, and R ３ and R ４ The compound of any one of claims 1 to 4, wherein together form an oxo (=O) group and Z is O.

14. The compound of claim 1, wherein R 1 is —H, R 2 is —OH, R 3 and R 4 are —H, and Z is —CH 2 —.

15. X and Y1 are independently —CH ２ The compound according to any one of claims 1 to 14, wherein the aryl group is - or -O-.

16. The compound according to any one of claims 1 to 14, wherein X and Y 1 are independently -CH 2 - or -S-. 【Request Item 17】 【Chemistry 8-1】 【Chemistry 8-2】 【Chemistry 8-3】 or a pharmaceutically acceptable salt thereof.

18. Lipidoid nanoparticles comprising a compound according to any one of claims 1 to 17.

19. 20. The lipidoid nanoparticle of claim 18, further comprising a protein, a nucleic acid, and / or a small molecule.

20. 20. The lipidoid nanoparticle of claim 19, comprising a protein, said protein being GFP-Cre.

21. 20. The lipidoid nanoparticle of claim 19, comprising a nucleic acid, wherein the nucleic acid is tau-ASO.

22. 20. The lipidoid nanoparticle of claim 19, comprising a small molecule, wherein the small molecule is an antifungal or chemotherapeutic agent.

23. and small molecules, such as 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, ruxolitinib, epirubicin, pirarubicin, idarubicin, valrubicin, amrubicin, bleomycin, phleomycin, dactinomycin, mithramycin, streptozotecin, pentostatin, and mitosin.

20. The lipidoid nanoparticle of claim 19, selected from the group consisting of mitomycin C, enediyne calicheamicin, glycoside rebeccamycin, macrolide lactone epothyphilone, ixabepilone, pentostatin, salinosporamide A, vinblastine, vincristine, etoposide, teniposide, vinorelbine, docetaxel, camptothecin, hycamtin, pederin, theopederin, annamide, trabectedin, aplidine, and ecteinascidin 743 (ET743).

24. 20. The lipidoid nanoparticle of claim 19, comprising a small molecule, wherein the small molecule is amphotericin B or doxorubicin.

25. Lipidoid nanoparticles according to any one of claims 18 to 24, having a particle size of 25 nm to 1000 nm.

26. A pharmaceutical composition comprising the lipidoid nanoparticles of any one of claims 18 to 25 and a pharmaceutically acceptable carrier or excipient.

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