Charge-altering releasable transporters from amino alcohol initiators

Abstract

The disclosure provides nucleic acid transporters in the form of copolymers derived from lipid carbonate monomers and cationic aminoester monomers prepared using various amino alcohol initiators, including linear and branched amino alcohol initiators, cyclic amino alcohol initiators and imidazole initiators. The compounds are useful for the delivery of nucleic acids to cells in vitro, ex vivo, or in vivo.

Description

CHARGE- AL TERING RELEASABLE TRANSPORTERS FROM AMINO ALCOHOLINITIATORS

[0001] This invention was made with Government support under contracts CHE— 2002933 awarded by the National Science Foundation and 5R01CA245533-03 awarded by the National Institutes of Health. The Government has certain rights in the invention.BACKGROUND

[0002] Nucleic acids are polyanionic, polar, and relatively large molecules on the order of about 20-200 kilodaltons (kDa) for plasmid DNA to between about 100-500 kDa for mRNA, as compared to small molecule therapeutics which are generally about 1 kilodalton or less. The large size and anionic charge of nucleic acids prevents or significantly hinders their unassisted passage through the nonpolar lipid membranes of cells and tissues. In addition, nucleic acids, especially RNAs, and particularly single-stranded mRNAs, are highly susceptible to enzymatic degradation. Two main strategies have developed to address these difficulties with delivery of nucleic acid based therapeutic agents. One approach is the development of noncharged and nonbiodegradable nucleic acid surrogates. Another approach is the development of delivery vehicles that facilitate delivery of the nucleic acids across lipid membranes. Delivery vehicles have included lipids, peptides, and aptamers as well as cationic polymers.

[0003] Efficient delivery vehicles for transporting nucleic acids into target cells and tissues, and where the nucleic acid encodes a polypeptide or protein, expression of the encoded polypeptide or protein, are needed to realize the full potential of nucleic acid-based technologies, including RNA-based vaccines and immunotherapies.BRIEF SUMMARY

[0004] The intracellular delivery of mRNA presents particular difficulties due both to its large size and amorphous secondary structure because, unlike plasmid DNA or siRNA, mRNA does not have a rigid double helix structure. The present invention provides new nucleic aciddelivery vehicles which have a demonstrated ability to deliver mRNA into cells where it is efficiently translated to provide high level protein expression both in vitro and in vivo.

[0005] Provided are compounds of Formula I or Formula la,Ri-X-Y-[(Li)xi-(L2)X2-(A2)y2-(Ai)yi]-R2 (Formula I)Ri-X-Y-[(Ai)yi-(A2)y2-(L2)x2-(Li)xi]-R2(Formula la) whereinRi is substituted or unsubstituted C2-C24 heteroalkyl or heterocycloalkyl, optionally substituted with one or more of hydroxyl, methyl, substituted or unsubstituted Ci-Cs alkyl, substituted 5 to 18 membered heterocycloalkyl or substituted 5 to 8 membered heterocycloalkenyl, wherein at least one heteroatom is N;X is a bond or linking group;Y is a bond, O, S, or N;Li and L2are each independently a lipid-functionalized methyl-trimethylene carbonate (MTC) unit having a structure ofwhere RL is a branched or unbranched, substituted or unsubstituted C1-C30 alkyl, which may be fully saturated, mono- or polyunsaturated;Ai and A2are each independently an aminoester monomer unit having a structure ofwherein RA is H, methyl, ethyl, aminoalkyl, substituted alkyl or substituted aryl, optionally substituted with thio, hydroxy, or cresyl; xi and yi are each independently greater than zero and less than 1,000;X2 and y2 are each independently zero or less than 1,000; andR2 is H or substituted or unsubstituted alkyl.

[0006] In aspects, Ri is the hydrocarbyl group of an amino alcohol initiator selected from AA01-AA22 or AA24-AA27.

[0007] In aspects, the compound may include where RL is branched, substituted C1-C30 alkyl, optionally substituted with a heteroaryl. In some aspects, RL is butyloctanyl, cholesteryl, dodecyl, linolenyl, linoleyl, nonenyl, oleyl, stearyl, or tocopheryl.

[0008] In aspects, the compound may include where RA is H, methyl, ethyl, -(CH2)3-NH2, - (CH2)4-NH2. -CH2-C6H4-OH or -(CH2)2-S-CH3 or tert-butoxy-methylbenzyl.

[0009] In aspects, the compound may include where R2 is substituted C2-C24 alkyl, optionally substituted with C2-C24 alkylcarbonyl, C1-C24 alkylcarbamoyl, carbamoyl, thiocarbamoyl, or C1-C24 thioalkylcarbamoyl.

[0010] Also provided are compounds where x2 and y2 are each zero and the compound is represented by Formula III or Formula Illa:Formula Illa wherein Ri, R2, xi, and yi are as defined for Formula I and Formula la, RLI is a branched or unbranched, substituted or unsubstituted C1-C30 alkyl, which may be fully saturated, mono- or polyunsaturated, and RAI is H, methyl, ethyl, aminoalkyl, substituted alkyl or substituted aryl, optionally substituted with thio, hydroxy, or cresyl.

[0011] In aspects, the compound may include where RLI is butyloctanyl, cholesteryl, dodecyl, linolenyl, linoleyl, nonenyl, oleyl, stearyl, or tocopheryl.

[0012] In aspects, the compound may include where RAI is H, methyl, ethyl, -(CH2)3-NH2, - (CH2)4-NH2, -CH2-C6H4-OH, -(CH2)2-S-CH3 or tert-butoxy-methylbenzyl. In some aspects, RAI is H.

[0013] In aspects, the compound may include where xl is 5-200 and yl is 5-100.

[0014] In aspects, provided is a compound selected from SC-3.05, SC-3.06, SC-5.03, SC-5.04, SC-6.01, SC-8.01, SC-8.03, SC-8.04, SC-9.04, SC-9.05, SC-10.00, SC-10.02, SC-11.00, SC- 11.04, SC-11.05, SC-11.08, SC-11.09, SC-11.11, SC-12.03, SC-12.04, SC-13.01, SC-13.02, SC-14.00 or SC-14.03.

[0015] Also provided are compositions and pharmaceutical compositions including nanoparticles of a compound of Formula I, la, II, Ila, III, or Illa, as described herein, non- covalently complexed with a nucleic acid or a plurality of different nucleic acids. The composition or pharmaceutical composition may also include a carrier or excipient. In aspects, the nucleic acid is RNA or DNA. In aspects, the nucleic acid is selected from messenger RNA (mRNA), small interference RNA (siRNA), short hairpin RNA (shRNA), micro RNA (miRNA), guide RNA (gRNA), CRISPR RNA (crRNA), transactivating RNA (tracrRNA), circular RNA (circRNA), self-amplifying RNA, plasmid DNA (pDNA), minicircle DNA, and genomic DNA (gNDA), and combinations of two or more of any of the foregoing.

[0016] The composition may also include where the nucleic acid is a therapeutic agent or the nucleic acid encodes a therapeutic agent.

[0017] The composition may also include where the composition is a vaccine.

[0018] Also provided are methods of transfecting a nucleic acid into a cell in vitro or ex vivo, the methods including contacting the cell in vitro or ex vivo with a composition or pharmaceutical composition including nanoparticles of a compound of Formula I, la, II, Ila, III, or Illa non-covalently complexed with the nucleic acid.

[0019] Also provided are methods of delivering a nucleic acid to a cell in a subject, the method including administering to the subject the composition or pharmaceutical composition including nanoparticles of a compound of Formula I, la, II, Ila, III, or Illa non-covalently complexed with the nucleic acid.

[0020] In aspects of the foregoing methods, the compound is SC-3.05, SC-3.06, SC-5.03, SC- 5.04, SC-6.01, SC-8.01, SC-8.03, SC-8.04, SC-9.04, SC-9.05, SC-10.00, SC-10.02, SC-11.00, SC-11.04, SC-11.05, SC-11.08, SC-11.09, SC-11.11, SC-12.03, SC-12.04, SC-13.01, SC-13.02, SC-14.00 or SC-14.03. In aspects, the compound is SC-6.01, SC-8.01, SC-9.04, SC-10.00, SC- 10.02, SC-11.04, SC-11.05, SC-11.08, SC-11.09, SC-12.03, SC-12.04 or SC-14.03. In aspects, the compound is BC-11, BC-13, BC-14, BC-15 or BC-16.

[0021] Also provided is a method of delivering a nucleic acid to a spleen of a subject, the method comprising administering to the subject a composition or pharmaceutical composition including a compound of Formula I, la, II, Ila, III, or Illa non-covalently complexed with the nucleic acid, wherein the compound is selected from SC-6.01, SC-8.01, SC-10.00, SC-11.00, SC-11.04, SC-11.05, SC-11.08, SC-12.04, SC-13.01, SC-13.02, or SC-14.00.

[0022] Also provided is a method of delivering a nucleic acid to a spleen or liver of a subject, the method comprising administering to the subject a composition or pharmaceutical composition including a compound of Formula I, Ta, IT, ITa, III, or Illa non-covalently complexed with the nucleic acid, wherein the compound is SC-11.05.

[0023] Also provided is a method of delivering a nucleic acid to a lung of a subject, the method comprising administering to the subject a composition or pharmaceutical composition including a compound of Formula I, la, II, Ila, III, or Illa non-covalently complexed with the nucleic acid, wherein the compound is selected from SC-2.00, SC-2.02, SC-3.01, SC-5.01, or SC-5.05.

[0024] Also provided is a method of delivering a nucleic acid to a liver of a subject, the method comprising administering to the subject a composition or pharmaceutical composition including a compound of Formula I, la, II, Tla, III, or Illa non-covalently complexed with the nucleic acid, wherein the compound is SC-9.02

[0025] Other technical features may be readily apparent to one skilled in the art from the following figures, descriptions, and claims.BRIEF DESCRIPTION OF THE FIGURES

[0026] FIG. 1 is a bar graph showing bioluminescence (total flux (p / s)) in mice (n=2) following intravenous injection via tail vein of fLuc mRNA complexed with compounds BC- ONA REF, BC-11, BC-12, BC-13, and BC-14 (left to right). BC-ONA REF was previously shown to support high levels of in vivo protein expression from complexed mRNA and is used as a reference compound. Also included as a reference compound is BC-12 which includes 1,3- propanediol as the initiator, rather than an amino alcohol initiator.

[0027] FIG. 2 is a bar graph showing bioluminescence (total flux (p / s)) in mice (n=2) following intravenous injection via tail vein of fLuc of mRNA complexed with compounds BC- 14, BC-15, BC-16, and SC-5.03 (left to right). The sequence of monomers is reversed in BC-15 as compared to BC-14, showing that both sequences are effective for in vivo protein expression. In contrast, BC-16, a high molecular weight block copolymer (DP=60) fails to show any protein expression while SC-5.03, a similar high molecular weight statistical copolymer (DP=68) showed unexpectedly high protein expression.

[0028] FIG. 3 is a bar graph showing bioluminescence (total flux (p / s)) in mice (n=2) following intravenous injection via tail vein of fLuc mRNA complexed with representativestatistical copolymers of Formula III. BC-ONA REF2 is used as a reference compound. Bioluminescence values are given in Tables 1.1-1.6, infra, which also give the architecture of the compounds.

[0029] FIG. 4A is a bar graph showing bioluminescence (total flux (p / s)) in mice (n=2) following intravenous injection via tail vein of fLuc mRNA complexed with compounds SC- 6.01, SC-8.01, SC-9.04, SC-10.02, SC-11.05, SC-12.03, and SC-14.03. BC-ONA REF2 is used as a reference.

[0030] FIG. 4B is a bar graph showing bioluminescence (total flux (p / s)) in mice (n=2) following intravenous injection via tail vein of fLuc mRNA complexed with compounds SC- 2.00, SC-2.02, SC-3.01, SC-5.01, and SC-5.05.

[0031] FIG. 5 is an image showing bioluminescence (total flux (p / s)) in mice (n=2) following intravenous injection via tail vein of fLuc mRNA complexed with compound SC-9.02, which showed high expression in the liver without expression in the spleen. Also shown is an image of two mice injected with fLuc mRNA complexed with a reference compound, BC-ONA REF2 (O6N6A9).

[0032] FIG. 6 illustrates structures of representative statistical copolymers of Formula III. Shown are compounds SC-11.00, SC-11.04, SC-11.05, and SC-11.08. Illustrated on the left side of the figure is AA22, the branched amino alcohol initiator moiety (Ri). The lipid / aminoester carbonate polymer moieties are designated as “Rp” at the initiation site on each arm of Ri in the AA22 structure. The different lipid / aminoester carbonate polymer moieties (Rp) for each compound are illustrated to the right of AA22 in the figure.

[0033] FIG. 7 illustrates structures of representative statistical copolymers of Formula III. Shown are compounds SC-6.01, SC-8.01 and SC-12.04. Illustrated on the left side of the figure are the branched amino alcohol initiator moieties for each compound (Ri), AA12, AA19, and AA23, respectively. The lipid / aminoester carbonate polymer moieties are designated as “Rp” at the initiation site on each arm of the AA structures. The different lipid / aminoester carbonate polymer moieties (Rp) for each compound are illustrated to the right of AA structures in the figure.

[0034] FIG. 8 illustrates structures of representative statistical copolymers of Formula III. Shown are compounds SC-13.01, SC-13.02, SC-10.00 and SC-14.00. Illustrated on the left side of the figure are the branched amino alcohol initiator moieties for each compound (Ri), AA24, AA21, and AA26, respectively. The lipid / aminoester carbonate polymer moieties aredesignated as “Rp” at the initiation site on each arm of the AA structures. The different lipid / aminoester carbonate polymer moieties (Rp) for each compound are illustrated to the right of AA structures in the figure.DETAILED DESCRIPTION

[0035] The disclosure provides compounds in the form of copolymers derived from lipid carbonate monomers and cationic aminoester monomers prepared using various amino alcohol initiators, including linear and branched amino alcohol initiators, cyclic amino alcohol initiators and imidazole initiators. The compounds disclosed herein are effective nucleic acid transporters as demonstrated by their ability to complex with mRNA and deliver the mRNA into cells where it is efficiently translated to produce high levels of protein expression in vitro and in vivo. In addition, certain compounds provide specific tropism to certain organs, such as spleen, lung and / or liver. Also provided are related compositions, including pharmaceutical compositions, and methods of delivering nucleic acids into cells, including delivery to cells in vitro, ex vivo, and in vivo.

[0036] Descriptions of compounds of the present disclosure are limited by principles of chemical bonding known to those skilled in the art. Accordingly, where a group may be substituted by one or more of a number of substituents, such substitutions are selected so as to comply with principles of chemical bonding and to give compounds which are not inherently unstable and / or would be known to one of ordinary skill in the art as likely to be unstable under ambient conditions, such as aqueous, neutral, and several known physiological conditions. For example, a heterocycloalkyl or heteroaryl is attached to the remainder of the molecule via a ring heteroatom in compliance with principles of chemical bonding known to those skilled in the art thereby avoiding inherently unstable compounds.

[0037] The term “alkyl,” by itself or as part of another substituent, refers to an acyclic branched or unbranched hydrocarbon group containing from about 1 to 24 carbon atoms (C1-C24) or from 1 to 18 carbon atoms (Ci-Cis), which may be fully saturated, mono- or polyunsaturated and can include mono-, di- and multivalent radicals. Examples of saturated alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, octadecyl, and the like. An unsaturated alkyl group is one having one or more double bonds or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(l,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and the higherhomologs and isomers. In some aspects, an unsaturated alkyl group may be a linolenyl or oleyl. “Substituted alkyl” refers to alkyl substituted with one or more substituent groups, including where two hydrogen atoms from the same carbon atom in an alkyl substituent are replaced, such as in a carbonyl group. For example, a substituted alkyl group may include a carbonyl (- C(=O)-) moiety. The term “heteroalkyl” refers to an alkyl substituent in which at least one carbon atom is replaced with a heteroatom, as described in more detail below. If not otherwise indicated, the term “alkyl” includes unsubstituted, substituted, and / or heteroatom-containing alkyl substituents.

[0038] The term “alkoxy” refers to an alkyl group bound through a single, terminal ether linkage which may be represented as -O-alkyl, where alkyl is as defined above. Examples include methoxy, ethoxy, n-propoxy, isopropoxy, t-butyloxy, etc. The term “alkylthio” refers to a group -S-alkyl.

[0039] The term “heteroalkyl”, which is shorthand for “heteroatom-containing alkyl”, refers to an alkyl substituent in which at least one carbon atom is replaced with a heteroatom. Similarly, the term “heterocyclic” refers to a cyclic substituent that is heteroatom-containing, and the term “heteroaryl” refers to “aryl” substituents that are heteroatom-containing. In this context, “heteroatom-containing” refers to a molecule, linkage or substituent in which one or more carbon atoms are replaced with an atom other than carbon, such as nitrogen (N), oxygen (O), sulfur (S), phosphorus (P) or silicon (Si), and more typically in the context of the present disclosure with N, O, or S. Examples of heteroalkyl groups include alkoxyaryl, alkylsulfanylsubstituted alkyl, N-alkylated amino alkyl, and the like. Examples of heteroaryl groups are provided below.

[0040] In some aspects, the heteroatom is O, N, or S. Examples include, but are not limited to: -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2- S-CH2, -S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -CH2-CH=N-OCH3, -CH=CH- N(CH3)-CH3, -O-CH3, -O-CH2-CH3, and -CN. Up to two or three heteroatoms may be consecutive, such as, for example, -CH2-NH-OCH3, R-S-S-R’, and RO-S(O)x-OR’. In some aspects, a heteroalkyl moiety may include one, two, three, four, or five heteroatom (e.g., O, N, S). In some aspects, a heteroalkyl moiety may include up to 8 optionally different heteroatoms (e.g., O, N, S).

[0041] The term “alkenyl” refers to a linear or branched hydrocarbon group of 2 to 24 carbon atoms containing at least one double bond, such as ethenyl, n-propenyl, isopropenyl, n-butenyl,isobutenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, octadecenyl, eicosenyl, and the like. In some aspects, alkenyl groups may contain 2 to about 18 carbon atoms (C2-C18) or 2 to 12 carbon atoms (C2-C12). The term “substituted alkenyl” refers to alkenyl substituted with one or more substituent groups, and the term “heteroalkenyl” refers to alkenyl in which at least one carbon atom is replaced with a heteroatom. A heteroalkenyl may optionally include more than one double bond and / or one or more triple bonds in additional to the one or more double bonds.

[0042] The term “alkylene” refers to a di-radical alkyl group. Unless otherwise indicated, such groups include saturated hydrocarbon chains containing from 1 to 24 carbon atoms, which may be substituted or unsubstituted, may contain one or more alicyclic groups, and may be heteroatom-containing. In aspects, an alkylene substituent group may be methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), 2-methylpropylene (-CH2-CH(CH3)-CH2-), hexylene (-(CH2)e-), octylene (-(CH2)s-) and the like. Similarly, the terms “alkenylene”, “alkynylene”, “arylene”, “aralkylene”, and “alkarylene” refer to di-radical alkenyl, alkynyl, aryl, aralkyl, and alkaryl groups, respectively. In some aspects, the aralkylene group is substituted on the alkylene moiety or the arylene moiety (e.g. at carbons 2, 3, 4, or 6) with a functional group. In the context of the present disclosure, alkylene groups are utilized as linking groups. Accordingly, these and other di-radical groups may be referred to herein as “linkers”, “linker groups”, “linker substituents”, “linking groups” or “linking substituents”. The alkylene, alkenylene, alkynylene, arylene, aralkylene, and alkarylene groups may also contain one or more functional groups. The term “functional group” in this context refers to di-radical moieties that contain one or more functional groups such as an oxo (-O-, such as in an ether linkage), amine (-NR-), carbonyl (-C(=O)-), carbonate, and the like. In some aspects, the functional group of a functional linker may be selected from oxo, -N3, -CN, -CHO, -OH, -NH2, -COOH, -CONH2, -NO2, -SH, -SO2CH3 -SO3H, , -OSO3H, -SO2NH2, -NHNH2, -ONH2, -NHC(0)NHNH2, substituted or unsubstituted C1-C5 alkyl or substituted or unsubstituted 2 to 5 membered heteroalkyl. In embodiments, the alkylarylene is unsubstituted.

[0043] The term "amino" is used herein to refer to the group -NZ1Z2 wherein Zi and Z2 are hydrogen or nonhydrogen substituents, with nonhydrogen substituents including, for example, alkyl, aryl, alkenyl, aralkyl, and substituted and / or heteroatom-containing variants thereof.

[0044] The terms “cycloalkyl” and “heterocycloalkyl” refer to cyclic versions of the “alkyl” and “heteroalkyl” groups defined above. Cycloalkyl and heterocycloalkyl are not aromatic. Inaspects, a cycloalkyl group includes monocyclic hydrocarbon ring systems containing from 3 to 9 carbon atoms (C3-C9), one or more of which may be replaced with a heteroatom, and where such groups can be saturated or unsaturated, but not aromatic. The term “3 to 6 membered” in reference to a cycloalkyl or heterocycloalkyl refers to the number of atoms, i.e. carbon atoms or carbon and one or more heteroatoms, in the monocyclic ring. For example, the term “3 to 6 membered cycloalkyl or heterocycloalkyl” refers to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, heterocyclopropyl, heterocyclobutyl, heterocyclopentyl, and heterocyclohexyl. In aspects, a cycloalkyl group includes a bicyclic or multi cyclic cycloalkyl ring system where multiple rings are fused together, where at least one of the fused rings is a cycloalkyl ring and the multiple rings are attached to the parent molecular moiety through any carbon atom contained within a cycloalkyl ring of the multiple rings. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1 -cyclohexenyl, 3- cyclohexenyl, cycloheptyl, and the like. Additionally, for heterocycloalkyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Examples of heterocycloalkyl include, but are not limited to, 1-(1, 2,5,6- tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1- piperazinyl, 2-piperazinyl, and the like. A “cycloalkylene” and a “heterocycloalkylene,” alone or as part of another substituent, means a divalent radical derived from a cycloalkyl and heterocycloalkyl, respectively. In embodiments, the cycloalkyl is fully saturated. In aspects, the cycloalkyl is monounsaturated. In embodiments, the cycloalkyl is polyunsaturated. In aspects, the heterocycloalkyl is fully saturated. In embodiments, the heterocycloalkyl is monounsaturated. In aspects, the heterocycloalkyl is polyunsaturated.

[0045] The term “acyl” means, unless otherwise stated, -C(O)R where R is a substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

[0046] The term “aryl” refers to cyclic groups that contain at least one aromatic ring, for example a single ring (e.g. phenyl) or multiple condensed rings (e.g. naphthyl). In some aspects of the present disclosure, the aryl group contains 6, 9 or 10 atoms such as phenyl, biphenyl, naphthyl, diphenylether, diphenylamine, benzophenone, indanyl, anthracenyl, 1,2- dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indanyl, indenyl and the like. A fusedring aryl refers to multiple rings fused together wherein at least one of the fused rings is an aryl ring and wherein the multiple rings are attached to the parent molecular moiety through any carbon atom contained within an aryl ring of the multiple rings. “Substituted aryl” refers to an aryl moiety substituted with one or more substituent groups, and the term “heteroaryl” refers to an aryl substituent in which at least one carbon atom is replaced with a heteroatom, as described in more detail below. If not otherwise indicated, the term “aryl” includes unsubstituted, substituted, and / or heteroatom-containing aromatic substituents.

[0047] The term “aralkyl” refers to an alkyl group with an aryl substituent, and the term “alkaryl” refers to an aryl group with an alkyl substituent, where “alkyl” and “aryl” are as defined above. In general, aralkyl and alkaryl groups may contain from 6 to 30 carbon atoms. Aralkyl and alkaryl groups may, for example, contain 6 to 20 carbon atoms or from 6 to 12 carbon atoms.

[0048] The term “heteroaryl” refers to aryl groups (or rings) that contain at least one heteroatom such as N, O, or S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized. Thus, the term “heteroaryl” includes fused ring heteroaryl groups (i.e., multiple rings fused together wherein at least one of the fused rings is a heteroaromatic ring and wherein the multiple rings are attached to the parent molecular moiety through any atom contained within a heteroaromatic ring of the multiple rings). A heteroaryl group can be attached to the remainder of the molecule through a carbon or heteroatom. Non-limiting examples of aryl and heteroaryl groups include phenyl, naphthyl, pyrrolyl, pyrazolyl, pyridazinyl, triazinyl, pyrimidinyl, imidazolyl, pyrazinyl, purinyl, oxazolyl, isoxazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrimidyl, benzothiazolyl, benzoxazoyl benzimidazolyl, benzofuran, isobenzofuranyl, indolyl, isoindolyl, benzothiophenyl, isoquinolyl, quinoxalinyl, quinolyl, 1 -naphthyl, 2-naphthyl, 4-biphenyl, 1 -pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4- oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5- thiazolyl, 2-furyl, 3 -furyl, 2-thienyl, 3 -thienyl, 2-pyridyl, 3 -pyridyl, 4-pyridyl, 2-pyrimidyl, 4- pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1 -isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituents for each of the above noted aryl and heteroaryl ring systems are selected from the group of acceptable substituents as described herein.

[0049] An “arylene” and a “heteroarylene,” alone or as part of another substituent, means a divalent radical derived from an aryl and heteroaryl, respectively. A heteroaryl group substituent may be -O- bonded to a ring heteroatom nitrogen.

[0050] For brevity, the term “aryl” when used in combination with other terms (e.g., aryloxy, arylthioxy, arylalkyl) includes both aryl and heteroaryl rings as defined above. Thus, the term “arylalkyl” is meant to include those radicals in which an aryl group is attached to an alkyl group (e.g., benzyl, phenethyl, pyridylmethyl, and the like) including those alkyl groups in which a carbon atom (e.g., a methylene group) has been replaced by, for example, an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl, 3-(l-naphthyloxy)propyl, and the like).

[0051] The terms “halo” and “halogen” are used in the conventional sense to refer to a chloro, bromo, fluoro or iodo substituent.

[0052] The term “substituted” as in “substituted alkyl”, “substituted aryl”, and the like, refers to at least one hydrogen atom bound to a carbon or other atom that is replaced with one or more non-hydrogen substituents in the alkyl, aryl, or other moiety. The term “substituted or unsubstituted” preceding a list, as in “substituted or unsubstituted C1-C24 alkyl, Ci- C 24 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or alkylaryl” is intended to modify each member of the list, as in "“substituted or unsubstituted C1-C24 alkyl, substituted or unsubstituted C1-C24 heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or substituted or unsubstituted alkylaryl”. Examples of such substituents include, without limitation: functional groups such as halo, hydroxyl, sulfhydryl, C1-C24 alkoxy, C2-C24 alkenyloxy, C5-C20 aryloxy, acyl (including C2-C24 alkylcarbonyl (-CO-alkyl) and C6-C20 arylcarbonyl (-CO-aryl)), acyloxy (-O-acyl), C2-C24 alkoxycarbonyl (-(CO)-O-alkyl), C6-C20 aryloxycarbonyl (-(CO)-O-aryl), halocarbonyl (-CO)-X where X is halo), C2-C24 alkylcarbonato (-O-(CO)-O-alkyl), C6-C20 arylcarbonato (-O-(CO)-O-aryl), carboxy (-COOH), carboxylato (-COO ), carbamoyl (-(CO)-NH2), mono-substituted C1-C24 alkylcarbamoyl (- (CO)-NH(CI-C24 alkyl)), di-substituted alkylcarbamoyl (-(CO)-N(CI-C24 alkyl)2), monosubstituted arylcarbamoyl (— (CO)— NH-aryl), thiocarbamoyl (-(CS)-NH2), carbamido (-NH- (CO)-NH2), formyl (-(CO)-H), thioformyl (-(CS)-H), amino (-NH2), mono- and di-(Ci-C24 alkyl)- substituted amino, mono- and di-(Cs-C2o aryl)-substituted amino, C2-C24 alkylamido (- NH-(CO)-alkyl), C5-C20 arylamido (-NH-(CO)-aryl), imino (-CR=NH where R is hydrogen, Ci- C24 alkyl, C5-C20 aryl, C6-C20 alkaryl, C6-C20 aralkyl, etc.), alkylimino (-CR=N(alkyl), where Ris hydrogen, alkyl, aryl, alkaryl, etc.), arylimino (-CR=N(aryl), where R is hydrogen, alkyl, aryl, alkaryl, etc.), nitro (-NO2), nitroso (-NO), sulfo (-SO2-OH), sulfonato (-SO2-O ), sulfonamide (-SO2NH), C1-C24 alkylsulfanyl (-S-alkyl or “alkylthio”), arylsulfanyl (-S-aryl or “arylthiol”), C1-C24 alkylsulfinyl (-(SO)-alkyl), C5-C20 arylsulfinyl (-(SO)-aryl), C1-C24 alkylsulfonyl (-SO2-alkyl), C5-C20 arylsulfonyl (-SO2-aryl); and the hydrocarbyl moieties Ci- C24 alkyl (including Ci-Cis alkyl, C1-C12 alkyl, Ci-Cs alkyl and Ci-Ce alkyl), C2-C24 alkenyl (including C2-C18 alkenyl, C1-C12 alkenyl, Ci-Cs alkenyl, and Ci-Ce alkenyl), C5-C30 aryl (including C5-C20 aryl, C5-C12 aryl), and C6-C30 aralkyl (including C6-C20 aralkyl, and C6-C12 aralkyl). In addition, the aforementioned functional groups may, if a particular group permits, be further substituted with one or more additional functional groups or with one or more hydrocarbyl moieties such as those specifically enumerated above. Analogously, the above- mentioned hydrocarbyl moieties may be further substituted with one or more functional groups or additional hydrocarbyl moieties such as those specifically enumerated. In addition, the hydrocarbyl moieties may contain one or more heteroatoms, optionally N, O, or both.

[0053] The suffix “ene” added on to any of the above groups means that the group is divalent, i.e. inserted between two other groups.

[0054] The symbol ” denotes the point of attachment of a chemical moiety to the remainder of a molecule or chemical formula.

[0055] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art. See, e.g., Singleton et al., Dictionary of Microbiology and Molecular Biology, 2nd ed., J. Wiley & Sons (New York, NY 1994); Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Springs Harbor Press (Cold Springs Harbor, NY 1989). Any methods, devices and materials similar or equivalent to those described herein can be used in the practice of this disclosure. The following definitions are provided to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure.Compounds

[0056] Provided are compounds in the form of statistical or block copolymers derived from two different types of monomers, lipid carbonate monomers and cationic aminoester monomers. The corresponding units of the copolymers derived from these monomers are referred to as “constitutional units” or “monomer units” or simply “units”. In some aspects, twodifferent types of lipid carbonate monomer unit and / or cationic aminoester monomer unit may be included in the copolymer such that the copolymers described here may comprise, e.g., from 2-4 different types of constitutional units derived from 2-4 different types of monomer.

[0057] Below, the statistical or block copolymers provided herein may be referred to using the nomenclature poly(A-stat-B) or polyA-block-polyB to differentiate between the statistical or block arrangement of the monomers, in accordance with Hodge et al., Pure Appl. Chem. 2020 92(5):797-813.

[0058] The polymer formulas described herein are presented as graphical representations of an “ideal” form of the polymer. For example, while the graphical representations may present the repeating monomer units as connected to each other in the same orientation, e.g., “head-to- tail”, it is understood that in the actual polymer there may also be some “head-to-head” and / or “tail-to-tail” dyads.

[0059] Provided are compounds of Formula I or Formula la:Ri-X-Y-[(Li)xi-(L2)x2-(A2)y2-(Ai)yi]-R2 (Formula I).Ri-X-Y-[(Ai)yi-(A2)y2-(L2)x2-(Li)xi]-R2 (Formula la). whereinRi is substituted or unsubstituted C2-C24 heteroalkyl or heterocycloalkyl, optionally substituted with one or more of hydroxyl, methyl, substituted or unsubstituted Ci-Cs alkyl, substituted 5 to 18 membered heterocycloalkyl or substituted 5 to 8 membered heterocycloalkenyl, wherein at least one heteroatom is N;X is a bond or a linking group as defined herein;Y is a bond, O, S, or N;Li and L2 are each independently a lipid carbonate monomer unit;Ai and A2 are each independently an aminoester monomer unit; xl and yl are each independently greater than zero and less than 1,000; x2 and y2 are each independently zero or less than 1,000; andR2 is H or substituted or unsubstituted alkyl.

[0060] In aspects, Li and L2 are each independently a lipid-functionalized methyltrimethylene carbonate (MTC) unit having a structure ofwhere RL is a branched or unbranched, substituted or unsubstituted C1-C30 alkyl, which may be fully saturated, mono- or polyunsaturated. In some aspects, Rtis branched, substituted C1-C30 alkyl, optionally substituted with a heteroaryl. In some aspects, RL is butyloctanyl, cholesteryl, dodecyl, linolenyl, linoleyl, nonenyl, oleyl, stearyl, or tocopheryl. In some aspects, RL is tocopheryl.

[0061] Ai and A2 are each independently an alpha aminoester (AAE) monomer unit having a structure ofwherein RA is H, methyl, ethyl, aminoalkyl, substituted alkyl, or substituted aryl, including thio substituted alkyl, hydroxy substituted aryl, and cresyl.

[0062] In some aspects, RA is H, methyl, ethyl, -(CH2)3-NH2, -(CH2)4-NH2, -CH2-C6H4-OH or -(CH2)2-S-CH3.

[0063] In aspects, xl, x2, yl, and y2 are each independently 5-1,000, 5-800, 5-600, 5-300, 5- 200, 5-100, 5-50, or 5-25. In some aspects, xl, x2, yl, and y2 are each independently 50-1,000, 50-900, 50-800, or 50-600. In some aspects, xl , x2, yl , and y2 are each independently 50-500, 50-400, 50-300, 50-200, or 50-100. In some aspects, xl, x2, yl, and y2 are each independently 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100. In some aspects, xl, x2, yl, and y2 are each independently 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100. In some aspects, X2 or Y2, or both, is zero.

[0064] In aspects, the sum of xl, x2, yl, and y2 is from 5-1,000, 5-800, 5-600, 5-300, 5-200, or 5-100. In some aspects, the sum of xl, x2, yl, and y2 is from 50-1,000, 50-800, 50-600, 50- 300, 50-200, or 50-100. In some aspects, the sum of xl, x2, yl, and y2 is from 20-30, 20-40, 20-50, 20-60, 20-70, 20-80, or 20-100. In some aspects, the sum of xl, x2, yl, and y2 is from 20-40 or from 20-100 or from 20-200.

[0065] In some aspects,RL isRA is H, C2-C10 aminoalkyl, alkylaryl, or alkoxy substituted phenyl; andRi is substituted or unsubstituted C2-C24 heteroalkyl or heterocycloalkyl, optionally substituted with one or more of hydroxyl, methyl, substituted or unsubstituted Ci-Cs alkyl, substituted 5 to 18 membered heterocycloalkyl or substituted 5 to 8 membered heterocycloalkenyl, wherein at least one heteroatom is N.

[0066] In some aspects, RL is oleyl, dodecyl, tocopheryl, or butyloctanyl; RA is H, C2-C10 aminoalkyl, alkylaryl, or alkoxy substituted phenyl; and Rl is C2-C6 substituted or unsubstituted aminoalkoxy, diaminoalkoxy, hydroxyaminoalkoxy, or dihydroxyaminoalkoxy, optionally substituted with one or more of methyl, ethyl, propyl, isopropyl, diisopropyl, hydroxyl, hydroxymethyl, hydroxyethyl, hydroxypropyl.

[0067] In some aspects, Ri is the hydrocarbyl group of an amino alcohol initiator selected from AA01-AA05:

[0068] In some aspects, Ri is the hydrocarbyl group of an amino alcohol initiator selected from AA06-AA10, AA19, AA20, or AA21 :

[0069] In some aspects, Ri is the hydrocarbyl group of an amino alcohol initiator selected from AA11, AA12, AA22, or AA27:

[0070] In some aspects, RL is butyloctanyl, dodecyl, oleyl, or tocopheryl;RA is H, C2-C10 aminoalkyl, alkylaryl, or alkoxy substituted phenyl; andRi is substituted pyrrolidinyl, piperazindiyl, diazepandiyl, imidazolyl, triazolyl, azacyclooctadecyl or diazacyclooctadecyl, optionally substituted with one or more of hydroxymethyl, hydroxy ethyl, hydroxypropyl, aminoalkoxy, or aminodialkoxy.

[0071] In some aspects, Ri is the hydrocarbyl group of an amino alcohol initiator selected from AA13-AA18 or AA24-AA26:

[0072] In accordance with each of the foregoing aspects where RA is H, C2-C10 aminoalkyl, alkylaryl, or alkoxy substituted phenyl, RA may be -(CH2)3NH2, -(CH2)4NH2, or tert-butoxy-methylbenzyl.

[0073] In accordance with any of the foregoing aspects, R2 may be substituted C2-C24 alkyl, optionally substituted with C2-C24 alkylcarbonyl, C1-C24 alkyl carbamoyl, carbamoyl, thiocarbamoyl, or C1-C24 thioalkylcarbamoyl.

[0074] In some aspects, the compound is represented by Formula II or Ila:Formula Ila where Ri, R2, xl, x2 and yl are as defined above; RLI and RL2 are each independently as defined for Ri, above; and RAI is as defined for RA above.

[0075] In some aspects, the compound is represented by Formula III or Illa:Formula Illa where Ri, R2, xl, and yl are as defined above or as defined in Tables 1-4; RLI is as defined above for RL or as defined in Tables 1-4; and RAI is as defined above for RA or as defined in Tables 1-4.

[0076] Table 1.1 : Representative statistical copolymers of Formula III and in vivo protein expression following intravenous delivery of compounds complexed with fLuc mRNA, measured as bioluminescence in units of flux (p / s).#Compound Ri RLI xl RAI y 1 R2 flux unitsSC-1.00 AA07 oleyl 8 H 8 H 16 6.08E+07(CH2)3-SC-2.00 AA08 butyloctanyl 10 3 H 13 1.31E+08NH2(CH2)3-SC-2.01 AA08 dodecyl 13 5 H 18NH2(CH2)3-SC-2.02 AA08 oleyl 14 6 H 20 3.78E+08NH2SC-3.00 AA09 butyloctanyl 14 H 15 H 29 9.77E+07(CH2)3-SC-3.01 AA09 butyloctanyl 18 9 H 27 1.00E+08NH2CH2-SC-3.02 AA09 butyloctanyl 22 C6H4- 5 H 27 1.05E+07OH(CH2)3-SC-3.03 AA09 dodecyl 18 5 H 23NH2CH2-SC-3.04 AA09 dodecyl 24 C6H4- 29 H 53OHCH2-SC-3.05 AA09 oleyl 19 C6H4- 5 H 24 5.61E+08OHSC-3.06 AA09 oleyl 30 H 30 H 60 5.67E+08(CH2)3-SC-3.07 AA09 oleyl 34 21 H 55 7.07E+06NH2SC-3.08 AA09 oleyl 13 H 11 H 24SC-3.09 AA09 tocopheryl 18 H 18 H 36 2.90E+07CH2-SC-3.10 AA09 tocopheryl 19 C6H4- 4 H 23 1.86E+06OHSC-4.00 AA10 butyloctanyl 8 H 9 H 17 2.70E+06SC-4.01 AA10 dodecyl 9 H 9 H 18 4.13E+07SC-4.02 AA10 oleyl 13 H 13 H 26 4.88E+07

[0077] Table 1.2 Representative statistical copolymers of Formula III (cont ).#Compound Ri RLI xl RAI yl R2 flux unitsSC-5.00 AA11 butyloctanyl 21 H 16 H 37 2.94E+07(CH2)3-SC-5.01 AA11 dodecyl 39 18 H 57 3.94E+07NH2SC-5.02 AA11 dodecyl 14 H 14 H 28SC-5.03 AA11 oleyl 35 H 33 H 68 1.35E+09SC-5.04 AA11 oleyl 12 H 8 H 20 1.29E+09(CH2)3-SC-5.05 AA11 oleyl 17 9 H 26 2.76E+07NH2SC-5.06 AA11 tocopheryl 20 H 12 H 32 1.08E+07SC-6.00 AA12 butyloctanyl 16 H 15 H 31 2.88E+08SC-6.01 AA12 dodecyl 18 H 16 H 34 7.23E+08SC-6.02 AA12 dodecyl 25 H 21 H 46SC-6.03 AA12 dodecyl 55 H 49 H 104SC-6.04 AA12 oleyl 18 H 16 H 34 2.57E+08SC-6.05 AA12 tocopheryl 35 H 34 H 69 1.73E+06

[0078] Table 1.3: Representative statistical copolymers of Formula III (cont.).#Compound Ri RLI xl RAI yl R2 flux unitsSC-7.00 AA18 butyloctanyl 16 H 14 H 30 4.31E+07SC-7.01 AA18 dodecyl 7 H 7 H 14 7.18E+06SC-8.00 AA19 butyloctanyl 11 H 10 H 21 2.35E+08SC-8.01 AA19 dodecyl 13 H 12 H 25 1.01E+09SC-8.02 AA19 dodecyl 19 H 14 H 33SC-8.03 AA19 dodecyl 25 H 22 H 47 5.55E+08SC-8.04 AA19 dodecyl 24 H 20 H 44 5.33E+08SC-8.05 AA19 oleyl 11 H 10 H 21 2.86E+08SC-8.06 AA19 tocopheryl 23 H 23 H 46 1.35E+07SC-9.00 AA20 butyloctanyl 14 H 12 H 26 3.63E+08(CH2)3-SC-9.01 AA20 dodecyl 12 4 H 16 2.45E+08NH2CH2-C6H4-SC-9.02 AA20 dodecyl 8 2 H 10 4.01E+08OHSC-9.03 AA20 dodecyl 9 H 8 H 17 4.99E+06CH2-C6H4-SC-9.04 AA20 oleyl 12 8 H 20 8.17E+08OHSC-9.05 AA20 oleyl 14 H 12 H 26 4.58E+08SC-9.06 AA20 tocopheryl 9 H 6 H 15 4.72E+06

[0079] Table 1.4: Representative statistical copolymers of Formula III (cont.).Compound Ri RLI xl RAI yl R2 # units fluxSC-10.00 AA21 butyloctanyl 12 H 10 H 22 7.87E+08SC-10.01 AA21 dodecyl 12 H 12 H 24 7.60E+07SC-10.02 AA21 oleyl 11 H 10 H 21 9.52E+08SC-10.03 AA21 oleyl 14 H 14 H 28SC-10.04 AA21 oleyl 29 H 26 H 55SC-10.05 AA21 tocopheryl 12 H 12 H 24 4.33E+07SC-11.00 AA22 butyloctanyl 19 H 18 H 37 5.79E+08SC-11.01 AA22 butyloctanyl 21 H 16 H 37 3.58E+08SC- 11.02 AA22 butyloctanyl 20 H 15 H 35SC-11.03 AA22 butyloctanyl 30 H 24 H 54SC-11.04 AA22 dodecyl 18 H 15 H 33 9.09E+08SC-11.05 AA22 dodecyl 20 H 20 H 40 2.11E+09SC-11.06 AA22 dodecyl 23 H 16 H 39SC-11.07 AA22 dodecyl 41 H 33 H 74SC-11.08 AA22 oleyl 16 H 12 H 28 8.02E+08SC-11.09 AA22 oleyl 18 H 17 H 35 1.23E+09SC-11.10 AA22 oleyl 20 H 15 H 35SC-11.11 AA22 oleyl 23 H 19 H 42 4.54E+08SC-11.12 AA22 oleyl 15 H 11 H 26SC-11.13 AA22 oleyl 34 H 32 H 66SC-11.14 AA22 tocopheryl 13 H 11 H 24

[0080] Table 1. : Representative statistical copolymers of Formula III (cont.).Compound Ri RLI xl RAI yl R2 # units fluxSC-12.00 AA23 butyloctanyl 14 H 10 H 24 1.62E+08SC-12.01 AA23 butyloctanyl 17 H 7 H 24 1.45E+07SC-12.02 AA23 dodecyl 15 H 14 H 29 2.48E+08SC-12.03 AA23 dodecyl 16 H 6 H 22 7.55E+08SC-12.04 AA23 oleyl 22 H 19 H 41 1.29E+09SC-12.05 AA23 oleyl 18 H 17 H 35SC-12.06 AA23 oleyl 25 H 21 H 46SC-12.07 AA23 tocopheryl 47 H 22 H 69 9.59E+05SC-13.00 AA24 butyloctanyl 14 H 10 H 24 6.39E+07SC-13.01 AA24 dodecyl 15 H 10 H 25 1.05E+08SC-13.02 AA24 oleyl 14 H 11 H 25 1.36E+08

[0081] Table 1.6: Representative statistical copolymers of Formula III (cont.).#Compound Rixl RAI yl R2 flux unitsSC-14.00 AA26 butyloctanyl 13 H 12 H 25 4.95E+08SC-14.01 AA26 butyloctanyl 27 H 25 H 52 1.99E+07SC-14.02 AA26 dodecyl 13 H 13 H 26 1.11E+08SC-14.03 AA26 oleyl 10 H 10 H 20 9.26E+08SC-14.04 AA26 tocopheryl 31 H 19 H 50 1.48E+06SC-15.00 AA27 butyloctanyl 142 H 61 H 200 5.35E+06SC-15.01 AA27 butyloctanyl 65 H 60 H 125 9.71E+06SC-15.02 AA27 dodecyl 37 H 35 H 72 6.45E+07SC-15.03 AA27 dodecyl 77 H 32 H 109 6.97E+06SC-15.04 AA27 oleyl 139 H 61 H 200 1.72E+07SC-15.05 AA27 oleyl 18 H 13 H 31 6.11E+07SC-15.06 AA27 tocopheryl 31 H 12 H 43 1.52E+07

[0082] FIG. 6 illustrates structures of representative statistical copolymers of Formula III, particularly compounds SC-11.00, SC-11.04, SC-11.05, and SC-11.08. Illustrated on the left side of the figure is AA22, the branched amino alcohol initiator moiety (Ri). The lipid / aminoester carbonate polymer moieties are designated as “Rp” at the initiation site on each arm of Ri in the AA22 structure. The different lipid / aminoester carbonate polymer moieties (Rp) for each compound are illustrated to the right of AA22 in the figure. The distribution of lipid / aminoester carbonate polymer moieties among the different arms of Ri was not determined but is approximated as the total number of lipid and aminoester monomer units (xl + yl) divided by the total number of initiation sites on the initiator moiety. For example, for compound SC-11.00, each of the 6 initiation sites is estimated to contain 19 / 6 lipid monomer units and 18 / 6 aminoester monomer units. FIG. 7 similarly illustrates compounds SC-6.01, SC- 8.01 and SC-12.04. FIG. 8 illustrates compounds SC-10.00, SC-13.01, SC-13.02 and SC-14.00.

[0083] Representative block copolymers of Formula Illa are given in Table 2. Table 3 gives the structures of the reference ONA compounds.

[0084] Table 2: Representative block copolymers of Formula Illa. “BDM” refers to benzene dimethanol, “MPAE” refers to 2-(methylphenylamino)ethanol and “PD” refers to 1,3-propanediol. Also shown is in vivo protein expression measured as flux (average of total) following intravenous delivery of compounds complexed with fLuc mRNA in mice (n=2).Compound Ri RLI xl RAI yl R2 # units fluxBC-8 (REF) benzyl oleyl 13 H 11 H 24 5.29E+08BC-9 (REF) BDM oleyl 13 H 11 H 24 3.41E+08BC-10 (REF) BDM oleyl 11 H 13 H 24 8.14E+08BC-11 MPAE oleyl 12 H 12 H 24 5.38E+08BC-12 (REF) PD oleyl 11 H 11 H 22 2.56E+08BC-13 AA08 oleyl 9 H 7 H 16 6.26E+08BC-14 AA11 oleyl 7 H 7 H 14 1.56E+09BC-15 (rev) AA11 oleyl 8 H 8 H 16 1.2E+09BC-16 AA11 oleyl 30 H 30 H 60 2.22E+07

[0085] Table 3: ONA reference compounds. Also shown is in vivo protein expression measured as flux (average of total) following intravenous delivery of compounds complexed with fLuc mRNA in mice (n=2).REF # x x RA y RCompoun Ri RLI RL2 unit flux1 2 1 1 2 d sBC-ONA benzy oley noneny 2.36E+06 6 H 8 H 20REFI 1 1 1 8BC-ONA benzy oley noneny 3.45E+06 6 H 9 H 21REF2 1 1 1 7Synthetic Methods

[0086] The synthesis of the compounds described herein proceeds according to scheme 1, in which an amino alcohol initiator (RiOH) is reacted with a lipid-derived MTC monomer and an amino acid ester (AAE) derived monomer to form a copolymer as shown in scheme 1. For monomer synthesis, see Blake, T. and Waymouth, R. M., “Organocatalytic Ring Opening Polymerization of Morpholinones: New Strategies to Functionalized Polyesters” J. Am. Chem. Soc., 2014, 136, 9252-9255; and / or McKinlay et al., “Charge-altering Releasable Transporters (CARTs) for the delivery and release of messenger RNA in living animals” Proc. Natl. Acad. Set., 2017, 114, E448-E456.where Ri, R2, RL and RA are defined as above. Exemplary amino alcohol initiators (RiOH) are provided by AA01-AA22 and AA24-AA27, as defined above.

[0087] In some aspects, the lipid-derived MTC monomer and the amino acid ester (AAE) derived monomer are independently selected from the following:Compositions

[0088] Also provided are compositions, including pharmaceutical compositions, comprising a compound as described herein and a carrier or excipient, including a pharmaceutically acceptable carrier or excipient.

[0089] In aspects, the composition or pharmaceutical composition comprises a compound of Formula I, II, or III, or subformula thereof, non-covalently complexed with a nucleic acid or a plurality of different nucleic acids. In aspects, the complexes condense to form nanoparticles ranging in size from 50-500 nanometers (nm) in diameter or from 100-300 nm. In this context, size refers to the mean particle size (Z). Thus, the disclosure also provides compositions or pharmaceutical compositions comprising nanoparticles of a compound of Formula I-III, orsubformula thereof, non-covalently complexed with a nucleic acid or a plurality of different nucleic acids.

[0090] In aspects, provided is a composition or pharmaceutical composition comprising a compound of Formula I, II, or III, or a subformula thereof, non-covalently complexed with a nucleic acid. In aspects, the complexes are in the form of nanoparticles. In aspects, the compound is a compound of Formula III. In aspects, the compound is SC-3.05, SC-3.06, SC- 5.03, SC-5.04, SC-6.01, SC-8.01, SC-8.03, SC-8.04, SC-9.04, SC-9.05, SC-10.00, SC-10.02, SC-11.00, SC-11.04, SC-11.05, SC-11.08, SC-11.09, SC-11.11, SC-12.03, SC-12.04, SC-13.01, SC-13.02, SC-14.00 or SC-14.03. In aspects, the compound is SC-6.01, SC-8.01, SC-9.04, SC- 10.00, SC-10.02, SC-11.04, SC-11.05, SC-11.08, SC-11.09, SC-12.03, SC-12.04 or SC- 14.03. In aspects, the compound is BC-11, BC-13, BC-14, BC-15 or BC-16. In aspects, the compound is SC-11.05.

[0091] In aspects, the nucleic acid is an RNA or DNA. In aspects, the nucleic acid is messenger RNA (mRNA), small interference RNA (siRNA), short hairpin RNA (shRNA), micro RNA (miRNA), guide RNA (gRNA), CRISPR RNA (crRNA), transactivating RNA (tracrRNA), circular RNA (circRNA), self-amplifying RNA, plasmid DNA (pDNA), minicircle DNA, and genomic DNA (gNDA), and combinations of two or more of any of the foregoing.

[0092] In aspects, the nucleic acid is a therapeutic agent. In aspects, the nucleic acid encodes a therapeutic agent.

[0093] In aspects, the nucleic acid includes one or more vectors, which may be eukaryotic expression vectors, bacterial plasmid vectors or viral vectors. In some aspects where the vector is a eukaryotic expression vector, the vector may include (a) a first polynucleotide encoding a CRISPR-Cas system guide RNA that hybridizes with a target sequence in the genome of eukaryotic cell, and (b) a second polynucleotide encoding a Cas9 protein, optionally wherein the Cas9 protein is codon optimized for expression in the cell. In aspects, the first (a) and second (b) polynucleotides are located in the same or different vectors. In aspects, the nucleic acid comprises a CRISPR RNA (crRNA), optionally wherein the crRNA is in the same plasmid vector as the first nucleotide sequence. In aspects, the nucleic acid comprises a transactivating RNA (tracrRNA). In aspects, the tracrRNA is optionally in the same vector as the second nucleotide sequence.

[0094] In aspects, the nucleic acid includes (a) a first polynucleotide encoding a transposase; and (b) a second polynucleotide comprising a nucleic acid sequence of a gene of interestflanked by a transposase recognition site. In aspects, the first (a) and second (b) polynucleotides are located in the same or different vectors. In accordance with this aspect, the transposase recognizes and excises a genomic sequence of interest.

[0095] In general, a compound of Formula I-III, or subformula thereof, is complexed with nucleic acid in an amount effective to produce a theoretical charge ratio of cationic compound to anionic nucleic acid of from about 2: 1 to about 30: 1 (cation: anion). In aspects, the theoretical charge ratio is 2: 1, 3: 1, 5: 1, 10: 1, 15: 1, 20: 1, 25: 1, or 30: 1. Theoretical (+ / -) charge ratios are calculated as moles of ammonium cations to moles of phosphate anions, assuming full amine protonation and phosphate deprotonation. In some aspects, the charge ratio (+ / -) is 3: 1, or 5: 1. Complexation of nucleic acid with a compound described herein may take place in the presence of a coordinating metal such as Zn+2, Mg+2, Ca+2; a dynamic non- covalent cross linker such as a carbohydrate; a counterion such as C1‘, AcO', succinate, or citrate; or a solubility modulator such as a lipid or a polyethyleneglycol (PEG), or any combination thereof.

[0096] In other aspects, the composition or pharmaceutical composition comprises a compound of Formula I-III, or a subformula thereof, without a nucleic acid cargo, for example where the nucleic acid is to be complexed at a later time such as before administration of the pharmaceutical composition to a subject.

[0097] In some aspects, the composition is a vaccine. In aspects where the composition is a vaccine, the composition may further include an immunological adjuvant. The immunological adjuvant can include, but is not limited to, agonists of Toll-like Receptors (TLRs), agonists of the STING pathway, agonistic antibodies against CD40, 0X40, CTLA4, PD1, or PD1-L, Freund’s adjuvant, bryostatins, PKC modulators, and ligands for CD40, 0X40, CD137, PD1, CTLA4 and any combinations thereof.

[0098] Pharmaceutically acceptable carriers or excipients include water, a pharmaceutically acceptable organic solvent, collagen, polyvinyl alcohol, polyvinylpyrrolidone, a carboxyvinyl polymer, carboxymethylcellulose sodium, polyacrylic sodium, sodium alginate, water-soluble dextran, carboxymethyl starch sodium, pectin, methyl cellulose, ethyl cellulose, xanthan gum, gum Arabic, casein, gelatin, agar, diglycerin, glycerin, propylene glycol, polyethylene glycol, Vaseline, paraffin, stearyl alcohol, stearic acid, human serum albumin (HSA), mannitol, sorbitol, lactose, a pharmaceutically acceptable surfactant and the like. Additional acceptable carriers, excipients, or stabilizers may include buffers such as phosphate, citrate, and otherorganic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG).

[0099] In some aspects, the composition or pharmaceutical composition may include a cryoprotectant agent. Non-limiting examples of cryoprotectant agents include a glycol (e.g., ethylene glycol, propylene glycol, and glycerol), dimethyl sulfoxide (DMSO), formamide, sucrose, trehalose, dextrose, and any combinations thereof.Methods of Use

[0100] The disclosure also provides methods for delivery of nucleic acids to cells and / or tissues in vitro, ex vivo, or in vivo. In aspects, the methods comprising contacting a target cell or tissue with a composition comprising nanoparticulate particles of a nucleic acid complexed with a compound of Formula I-III, or subformula thereof. In aspects, the methods comprise administering to a subject a composition comprising nanoparticulate particles of a nucleic acid complexed with a compound of Formula I-III, or subformula thereof, including a pharmaceutical composition or vaccine. Administration may be according to any suitable route, for example, parenteral, including e.g., intravenous, intramuscular, intradermal, subcutaneous, intraperitoneal, intracranial, etc., or transmucosal, including e.g., buccal, nasal, sublingual, transdermal, etc. In aspects, the subject is a mammal, for example a human, a non-human primate, a murine (i.e., mouse and rat), a canine, a feline, or an equine. In aspects, the subject is a human.

[0101] In an aspect, provided is a method of transfecting a nucleic acid into a cell in vitro or ex vivo, the method comprising contacting the cell in vitro or ex vivo with a compound of Formula I, II, or III, or a subformula thereof, non-covalently complexed with the nucleic acid, or a composition comprising a compound of Formula I, II, or III, or a subformula thereof, non-covalently complexed with the nucleic acid. In aspects, the complexes are in the form of nanoparticles. In aspects, the compound is SC-3.05, SC-3.06, SC-5.03, SC-5.04, SC-6.01, SC- 8.01, SC-8.03, SC-8.04, SC-9.04, SC-9.05, SC-10.00, SC-10.02, SC-11.00, SC-11.04, SC- 11.05, SC-11.08, SC-11.09, SC-11.11, SC-12.03, SC-12.04, SC-13.01, SC-13.02, SC-14.00 or SC-14.03. In aspects, the compound is SC-11.05.

[0102] In an aspect, provided is a method of delivering a nucleic acid to a cell in a subject, the method comprising administering to the subject the composition or pharmaceutical composition comprising non-covalent complexes of a compound of Formula I, II, or III, or a subformula thereof, with the nucleic acid. In aspects, the complexes are in the form of nanoparticles. In aspects, the compound is SC-3.05, SC-3.06, SC-5.03, SC-5.04, SC-6.01, SC-8.01, SC-8.03, SC- 8.04, SC-9.04, SC-9.05, SC-10.00, SC-10.02, SC-11.00, SC-11.04, SC-11.05, SC-11.08, SC- 11.09, SC-11.11, SC-12.03, SC-12.04, SC-13.01, SC-13.02, SC-14.00 or SC-14.03. In aspects, the compound is SC-11.05.

[0103] In an aspect, provided is a method of delivering a nucleic acid to the spleen of a subject, where the method includes administering to the subject a composition or pharmaceutical composition comprising non-covalent complexes of a compound of Formula I, II, or III, or a subformula thereof, with a nucleic acid or a plurality of different nucleic acids. In aspects, the complexes are in the form of nanoparticles. In aspects, the compound is SC-6.01, SC-8.01, SC-10, SC-11.00, SC-11.04, SC-11.05, SC-11.08, SC-12.04, SC-13.01, SC-13.02, or SC-14.00.

[0104] In an aspect, provided is a method of delivering a nucleic acid to the spleen of a subject, where the method includes administering to the subject a composition or pharmaceutical composition comprising non-covalent complexes of a compound of Formula I, II, or III, or a subformula thereof, with a nucleic acid or a plurality of different nucleic acids. In aspects, the complexes are in the form of nanoparticles. In aspects, the compound is SC-11.05.

[0105] In an aspect, provided is a method of delivering a nucleic acid to the liver of a subject, where the method includes administering to the subject a composition or pharmaceutical composition comprising non-covalent complexes of a compound of Formula I, II, or III, or a subformula thereof, with a nucleic acid or a plurality of different nucleic acids. In aspects, the complexes are in the form of nanoparticles. In aspects, the compound is SC-9.02.

[0106] In accordance with any of the foregoing methods, the nucleic acid may be RNA or DNA. In aspects, the RNA is messenger RNA (mRNA), small interference RNA (siRNA), shorthairpin RNA (shRNA), micro RNA (miRNA), guide RNA (gRNA), CRISPR RNA (crRNA), or transactivating RNA (tracrRNA). In aspects, the DNA is plasmid DNA (pDNA), minicircle DNA, or genomic DNA (gNDA).

[0107] In aspects, the nucleic acid may be a therapeutic nucleic acid or the nucleic acid may encode one or therapeutic agents, for example a cytokine, a T cell receptor (TCR), or a chimeric antigen receptor (CAR).

[0108] In some aspects, a method of transfecting a nucleic acid into a cell as described herein may be part of a method for gene editing or genetic engineering. For example, one or more nucleic acids may be transfected using the methods described herein in a CRISPR-based system or a transposon-based system for gene editing or genetic engineering. Accordingly, one or more nucleic acids may be transfected according to the methods described here, which nucleic acids may be located on one or more vectors. For example, the one or more nucleic acids may comprise a vector having a first nucleotide sequence encoding a CRISPR-Cas system guide RNA (gRNA) that hybridizes with a target sequence in the genome of a target cell and a second nucleotide sequence encoding a Cas9 protein. Alternatively, the gRNA and Cas9 protein can be located on different vectors, or either the gRNA or Cas9 protein may be produced in the target cell. The one or more nucleic acids may also comprise a CRISPR RNA (crRNA) and / or transactivating RNA (tracrRNA), each of which may be located on the same or a different vector as the gRNA and / or Cas9 encoding sequence.

[0109] In another aspect, the one or more nucleic acids may comprise a sequence encoding a transposase and a nucleic acid sequence of a gene of interest flanked by a transposase recognition site, which may be located on the same or different vectors.

[0110] In some aspects, a compound of Formula I, II, or III, or subformula thereof, may be complexed with one or more nucleic acids encoding one or more antigenic or immunogenic epitopes or peptides which may form a vaccine composition. In some aspects, a mixture of two or more different compounds of Formula I, II, or III, or subformula thereof, may be complexed with one or more nucleic acids encoding one or more antigenic or immunogenic epitopes or peptides.Biological Examples

[0111] The compounds described herein are able to form stable complexes with nucleic acids, including mRNA, which complexes are effective to transporting their nucleic acid cargo intocells in vivo following intravenous delivery. In addition, as evidenced by the examples described below, compounds within the scope of the present invention complexed with mRNA not only deliver the mRNA into cells following intravenous delivery, but are further able to escape the endosomal compartment and release the mRNA into the cytosol for translation resulting in high level protein expression. The successful high level expression of encoded proteins indicates that complexes formed with compounds of the present invention are adapted to avoid renal clearance, protect their nucleic acid cargo against enzymatic degradation, and avoid serum aggregation in the bloodstream. In addition, the compounds are biodegradable over relatively short time periods of less than 24 or 48 hours, thereby reducing the chance of cytotoxicity or immunogenicity related to the compounds themselves.

[0112] Representative compounds of Formula III and Illa showed unexpectedly high levels of in vivo protein expression when complexed with fLuc mRNA and injected into the tail vein of mice. FIG. 3 and Tables 1.1-1.6 show in vivo protein expression measured as fLuc bioluminescence for compounds of Formula III. Values are average total flux (n= 2 mice) measured 4 hours after tail-vein injection. As evidenced by the data, many compounds of Formula III and Illa showed remarkably high protein expression. As a reference, previously described compounds, BC-ONA REFI and BC-ONA REF2 (Table 3) provided average fluxes in the range of 3.45E+07 to 2.36 E+8 p / s, which is 1 to 2 orders of magnitude lower than that achieved for many of the compounds of Formula III and Illa.

[0113] Compounds exhibiting high protein expression, defined as a flux of at least about 5E+8 p / s, include SC-3.05, SC-3.06, SC-5.03, SC-5.04, SC-6.01, SC-8.01, SC-8.03, SC-8.04, SC- 9.04, SC-10.00, SC-10.02, SC-11.00, SC-11.04, SC-11.05, SC-11.08, SC-11.09, SC-12.03, SC- 12.04, SC-13.01, SC-13.02, SC-14.00 and SC-14.03.

[0114] The following compounds exhibited high protein expression in the spleen: SC-6.01, SC-8.01, SC-10.00, SC-11.00, SC-11.04, SC-11.05, SC-11.08, SC-12.04, SC-13.01, SC-13.02 and SC-14.00.

[0115] Compound SC-11.05 exhibited high protein expression in both spleen and liver.

[0116] The following compounds exhibited selective protein expression in the lung: SC-2.00, SC-2.02, SC-3.01, SC-5.01 and SC-5.05.

[0117] Compound SC-9.02 exhibited selective protein expression in the liver.

[0118] Compounds SC-6.01, SC-8.01, SC-9.04, SC-10.00, SC-10.02, SC-11.04, SC-11.05, SC-11.08, SC-11.09, SC-12.03, SC-12.04 and SC-14.03 all demonstrated particularly high levels of in vivo protein expression, defined as a flux of at least about 7E+8 p / s.

[0119] Compounds of Formula Illa exhibiting high protein expression include BC-11, BC-13, BC-14, BC-15 and BC-16.

[0120] FIG. 1 shows results of a representative experiment in which bioluminescence (total flux (p / s)) was measured in mice (n=2) following intravenous injection via tail vein of firefly luciferase (fLuc) mRNA complexed with compounds BC-ONA REF, BC-11, BC-12, BC-13, and BC-14 (left to right). BC-ONA REF was previously shown to support high levels of in vivo protein expression from complexed mRNA and is used as a reference. Also included as a reference compound is BC-12 which includes 1,3 -propanediol as the initiator, rather than an amino alcohol initiator.

[0121] FIG. 2 shows results of another representative experiment showing in vivo protein expression measured as bioluminescence (total flux (p / s)) in mice (n=2) following intravenous injection via tail vein of fLuc of mRNA complexed with compounds BC-14, BC-15, BC-16, and SC-5.03 (left to right). The sequence of monomers is reversed in BC-15 as compared to BC-14, showing that both sequences are effective for in vivo protein expression. In contrast, BC-16, a high molecular weight block copolymer (DP=60), fails to show any protein expression while SC-5.03, a similar high molecular weight statistical copolymer (DP=68) showed unexpectedly high protein expression.

[0122] FIG. 3 is a bar graph showing in vivo protein expression measured as bioluminescence (total flux (p / s)) in mice (n=2) following intravenous injection via tail vein of fLuc of mRNA complexed with additional compounds of Table 1.

[0123] FIG. 4A is a bar graph showing in vivo protein expression measured as bioluminescence (total flux (p / s)) in mice with some of the highest performing compounds, SC- 6.01, SC-8.01, SC-9.04, SC-10.02, SC-11.05, SC-12.03, and SC-14.03. BC-ONA REF2 is used as a reference.

[0124] FIG. 4B is a bar graph showing in vivo protein expression measured as bioluminescence (total flux (p / s)) in mice for five compounds that were lung-tropic, left to right: SC-5.05, SC-5.01, SC-3.01, SC-2.00 and SC-2.02.

[0125] FIG. 5 shows an image of in vivo protein expression measured as bioluminescence (total flux (p / s)) in mice for compound SC-9.02, which showed high expression in the liver without expression in the spleen.

[0126] In the foregoing description and the following claims, the following will be appreciated. The phrases “at least one”, “one or more”, and “and / or”, as used herein, are open- ended expressions that are both conjunctive and disjunctive in operation. The terms “a”, “an”, “the”, “first”, “second”, etc., do not preclude a plurality. For example, the term “a” or “an” entity, as used herein, refers to one or more of that entity. As such, the terms “a” (or “an”), “one or more” and “at least one” can be used interchangeably herein.

[0127] The abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical sciences.

[0128] The terms “oligomer” and “polymer” are used interchangeably herein to refer to a compound that has a plurality of repeating subunits, which may be referred to as blocks or monomer units, or simply as monomers. The terms “co-oligomer” or “copolymer” are used interchangeably herein to refer to an oligomer or polymer that includes two or more different types of monomers.

[0129] The term “initiator” refers to a compound that initiates a polymerization reaction. Thus, the initiator is typically incorporated at the end of the synthesized polymer. For example, a plurality of monomer molecules can be reacted with an initiator to provide a polymer or copolymer. The initiator can be present on at least one end of the resulting polymer or copolymer and not constitute a repeating monomer unit of the polymer.

[0130] The term “small organic molecule” refers to a small organic compound, including heteroorganic and organometallic compounds, having a molecular weight less than about 1,000 grams per mole, or less than about 500 grams per mole and salts, esters, and other pharmaceutically acceptable forms of such compounds.

[0131] While the invention herein disclosed has been described by means of specific embodiments and applications thereof, numerous modifications and variations could be made thereto by those skilled in the art without departing from the scope of the invention set forth in the claims.

[0132] It will be appreciated that the present invention is set forth in various levels of detail in this application. In certain instances, details that are not necessary for one of ordinary skill inthe art to understand the invention, or that render other details difficult to perceive may have been omitted. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting beyond the scope of the appended claims. Unless defined otherwise, technical terms used herein are to be understood as commonly understood by one of ordinary skill in the art to which the disclosure belongs.

[0133] In the claims, the term “comprises / comprising” does not exclude the presence of other elements, components, features, regions, integers, steps, operations, etc. Additionally, although individual features may be included in different claims, these may possibly advantageously be combined, and the inclusion in different claims does not imply that a combination of features is not feasible and / or advantageous. The transitional phrase “consisting essentially of’ (and grammatical variants) is to be interpreted as encompassing the recited materials or steps “and those that do not materially affect the basic and novel characteristic(s)” of the recited embodiment. Thus, the term “consisting essentially of’ as used herein should not be interpreted as equivalent to “comprising.”

Claims

CLAIMSWhat is claimed is:1 . A compound of Formula I or Formula la,Ri-X-¥-[(Li)xi-(L2)x2-(A2)y2-(Ai)yi]-R2 (Formula I)Ri-X-Y-[(Ai)yi-(A2)y2-(L2)x2-(Li)xi]-R2 (Formula la) whereinRi is substituted or unsubstituted C2-C24 heteroalkyl or heterocycloalkyl, optionally substituted with one or more of hydroxyl, methyl, substituted or unsubstituted Ci-Cs alkyl, substituted 5 to 18 membered heterocycloalkyl or substituted 5 to 8 membered heterocycloalkenyl, wherein at least one heteroatom is N;X is a bond or linking group;Y is a bond, O, S, or N;Li and L2 are each independently a lipid-functionalized methyl-trimethylene carbonate(MTC) unit having a structure ofwhere RL is a branched or unbranched, substituted or unsubstituted C1-C30 alkyl, which may be fully saturated, mono- or polyunsaturated;Ai and A2 are each independently an aminoester monomer unit having a structure ofwherein RA is H, methyl, ethyl, aminoalkyl, substituted alkyl or substituted aryl, optionally substituted with thio, hydroxy, or cresyl; xi and yi are each independently greater than zero and less than 1,000;X2 and y2 are each independently zero or less than 1,000; andR2 is H or substituted or unsubstituted alkyl.

2. The compound of claim 1, wherein Ri is the hydrocarbyl group of an amino alcohol initiator selected from AA01-AA22 or AA24-AA27.

3. The compound of claim 1 or 2, wherein Ri s branched, substituted C1-C30 alkyl, optionally substituted with a heteroaryl.

4. The compound of claim 1 or 2, wherein RL is butyloctanyl, cholesteryl, dodecyl, linolenyl, linoleyl, nonenyl, oleyl, stearyl, or tocopheryl.

5. The compound of any one of claims 1 to 4, wherein RA is H, methyl, ethyl, -(CH2)3-NH2, - (CH2)4-NH2, -CH2-C6H4-OH, -(CH2)2-S-CH3 or tert-butoxy-methylbenzyl.

6. The compound of any one of claims 1 to 5, wherein R2 is substituted C2-C24 alkyl, optionally substituted with C2-C24 alkylcarbonyl, C1-C24 alkylcarbamoyl, carbamoyl, thiocarbamoyl, or C1-C24 thioalkylcarbamoyl.

7. The compound of any one of claims 1 to 6, wherein x2 and y2 are each zero and the compound is represented by Formula III or Formula Illa:wherein Ri, R2, xi, and yi are as defined in any one of claims 1 to 6,RLI is a branched or unbranched, substituted or unsubstituted C1-C30 alkyl, which may be fully saturated, mono- or polyunsaturated, andRAI is H, methyl, ethyl, aminoalkyl, substituted alkyl or substituted aryl, optionally substituted with thio, hydroxy, or cresyl.

8. The compound of claim 7, wherein RLI is butyloctanyl, cholesteryl, dodecyl, linolenyl, linoleyl, nonenyl, oleyl, stearyl, or tocopheryl.

9. The compound of claim 7 or 8, wherein RAI is H, methyl, ethyl, -(CH2)3-NH2, -(CH2)4-NH2, - CH2-C6H4-OH, -(CH2)2-S-CH3 or tert-butoxy-methylbenzyl.

10. The compound of claim 8, wherein RAI is H.

11. The compound of any one of claims 1 to 9, wherein xi is 5-200 and yi is 5-100.

12. The compound of any one of claims 7-9 or 11, wherein the compound is SC-3.05, SC-3.06, SC-5.03, SC-5.04, SC-6.01, SC-8.01, SC-8.03, SC-8.04, SC-9.04, SC-9.05, SC-10.00, SC- 10.02, SC-11.00, SC-11.04, SC-11.05, SC-11.08, SC-11.09, SC-11.11, SC-12.03, SC-12.04, SC-13.01, SC-13.02, SC-14.00 or SC-14.03.

13. A composition or pharmaceutical composition comprising the compound of any one of claims 1-9 or 11-12 non-covalently complexed with a nucleic acid or a plurality of different nucleic acids.

14. The composition of claim 13, wherein the nucleic acid is RNA or DNA.

15. The composition of claim 13, wherein the nucleic acid is selected from messenger RNA (mRNA), small interference RNA (siRNA), short hairpin RNA (shRNA), micro RNA (miRNA), guide RNA (gRNA), CRISPR RNA (crRNA), transactivating RNA (tracrRNA), circular RNA (circRNA), self-amplifying RNA, plasmid DNA (pDNA), minicircle DNA, and genomic DNA (gNDA), and combinations of two or more of any of the foregoing.

16. The composition of any one of claims 13 to 15, wherein the nucleic acid is a therapeutic agent or the nucleic acid encodes a therapeutic agent.

17. The composition of any one of claims 13 to 16, wherein the composition is a vaccine.

18. A method of transfecting a nucleic acid into a cell in vitro or ex vivo, the method comprising contacting the cell in vitro or ex vivo with the composition or pharmaceutical composition of any one of claims 13 to 17.

19. A method of delivering a nucleic acid to a cell in a subject, the method comprising administering to the subject the composition or pharmaceutical composition any one of claims 13 to 17.

20. The method of claim 18 or 19, wherein the compound is SC-3.05, SC-3.06, SC-5.03, SC- 5.04, SC-6.01, SC-8.01, SC-8.03, SC-8.04, SC-9.04, SC-9.05, SC-10.00, SC-10.02, SC-11.00, SC-11.04, SC-11.05, SC-11.08, SC-11.09, SC-11.11, SC-12.03, SC-12.04, SC-13.01, SC-13.02, SC-14.00 or SC-14.03.

21. A method of delivering a nucleic acid to a spleen and / or a liver of a subject, the method comprising administering to the subject the composition or pharmaceutical composition any one of claims 13 to 17, wherein the compound is selected from SC-6.01, SC-8.01, SC-9.02, SCI O, SC-1 1.00, SC-1 1.04, SC-1 1.05, SC-1 1.08, SC-12.04, SC-13.01 , SC-13.02, or SC-14.00.

22. A method of delivering a nucleic acid to a lung of a subject, the method comprising administering to the subject the composition or pharmaceutical composition any one of claims 13 to 17, wherein the compound is selected from SC-2.00, SC-2.02, SC-3.01, SC-5.01, or SC- 5.05.

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