Backbone-degradable polymers, vectors comprising the same, and methods of use thereof

Abstract

The disclosure provides, in part, a polymer composition comprising a random copolymer of at least one hydrophilic vinyl monomer, at least one cationic vinyl monomer, and at least one cyclic monomer, vectors comprising the polymer composition of the disclosure and at least one nucleic acid, and methods of use thereof. In certain embodiments, the cyclic monomer of the disclosure is a compound of formula (Ia), which comprises at least one degradable moiety:

Description

[0001] Attorney Docket No. 370602-7086W01(00285)

[0002] TITLE OF THE INVENTION

[0003] Backbone-Degradable Polymers, Vectors Comprising the Same, and Methods of Use Thereof

[0004] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 U. S. C. § 119(e) to U. S. Provisional Patent Application No. 63 / 746,824. filed January 17, 2025, which is incorporated herein by reference in its entirety.

[0005] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0006] This invention was made with government support under grant number R35GM138296 awarded by the National Institutes of Health and grant number CHE 1944512 awarded by the National Science Foundation. The government has certain rights in the invention.

[0007] BACKGROUND

[0008] Viral gene delivery is the most widely studied approach to deliver genetic payloads into cells for gene therapy. However, concerns about safety7, immunogenicity7, and payload limitations have led to increased interest in developing alternative synthetic delivery vectors. Polymers in particular present an attractive alternative due to their significant synthetic flexibility that allows their chemical composition to be precisely tailored. The incorporation of cationic monomers into the polymer generates a highly charged macromolecule that can condense nucleic acid to form a polyelectrolyte complex. Such complexes can navigate across various cellular barriers to efficiently deliver the nucleic acid payload intracellularly. However, these highly charged polymers can also lead to cell cytotoxicity.

[0009] There is thus a need in the art for biocompatible and / or degradable polymers and methods of use thereof for effective gene delivery. The present disclosure addresses this need.

[0010] BRIEF SUMMARY OF THE INVENTION

[0011] In one aspect, the disclosure provides a polymer composition comprising a random copolymer of:

[0012] (a) at least one hydrophilic vinyl monomer, wherein the at least one hydrophilic vinyl monomer comprises a vinyl monomer substituted with at least one hydrophilicAttorney Docket No. 370602-7086W01(00285)

[0013] moiety;

[0014] (b) at least one cationic vinyl monomer, wherein the at least one cationic vinyl monomer comprises a vinyl monomer substituted with at least one cationic moiety; and

[0015] (c) at least one cyclic monomer of formula (la):

[0016] \ X1vL1

[0017] R2bAzAR

[0018]

[0019] R2aR1 b(la), wherein:

[0020] Rlaand Rlbare each independently selected from the group consisting of H, optionally substituted Ci-Ce alkyl, and optionally substituted Ce-Cio aryl;

[0021] R2aand R2bare each independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, and optionally substituted Cg-Cio aryl;

[0022] R3aand R3bare each independently H;

[0023] L1comprises at least one divalent moiety selected from the group consisting of optionally substituted C1-C12 alkylenyl, optionally substituted C1-C12 heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted Ce-Cio arylenyl, and optionally substituted C2-C10 heteroarylenyl;

[0024] X1is selected from the group consisting of -O-*, -S-*, -S-S-*, -C(=O)O-*, -OC(=O)-*, - NHC(=O)-*, -C(=O)NH-*, -C(=S)O-*, -OC(=S)-*, -OC(=O)O-*, -NHC(=S)S-*, -SC(=S)NH-*, -NHC(=O)O-*, -OC(=O)NH-*, and a bond (absent);

[0025] X2is selected from the group consisting of **-O-, **-S-, **-S-S-. **-C(=O)O-, **- OC(=O)-, **-NHC(=O)-, **-C(=O)NH-, **-€(=8)0-, **-OC(=S)-, **-OC(=O)O-, **-NHC(=S)S-, **-SC(=S)NH-, **-NHC(=O)O-, **-OC(=O)NH-, and a bond;

[0026] wherein no more than one of X1and X2is a bond;

[0027] Z1is selected from the group consisting of -S- and -S(=O)2-;

[0028] * indicates a bond between X1and L1; and

[0029] ** indicates a bond between X2and L1.

[0030] In certain embodiments, the disclosure provides a vector comprising the polymer composition of the disclosure and at least one nucleic acid cargo, wherein the polymer and nucleic acid cargo are non-covalently complexed.

[0031] In another aspect, the disclosure provides a pharmaceutical composition comprising the vector of the disclosure and at least one pharmaceutically acceptable excipient.

[0032] In another aspect, the disclosure provides a method of delivering a nucleic acid to aAttorney Docket No. 370602-7086W01(00285)

[0033] target cell, the method comprising administering to the subject comprising the target cell a vector of the disclosure and / or a pharmaceutical composition of the disclosure.

[0034] In another aspect, the disclosure provides a method for performing gene therapy in a subject, the method comprising administering to the subject comprising the target cell a vector of the disclosure and / or a pharmaceutical composition of the disclosure.

[0035] In another aspect, the disclosure provides a method for treating, ameliorating, and / or preventing a disease in a subject, the method comprising administering to the subject comprising the target cell a vector of the disclosure and / or a pharmaceutical composition of the disclosure.

[0036] In another aspect, the disclosure provides a method for modulating gene expression in a target cell, the method comprising administering to the subject comprising the target cell a vector of the disclosure and / or a pharmaceutical composition of the disclosure.

[0037] BRIEF DESCRIPTION OF THE FIGURES

[0038] The drawings illustrate generally, by way of example, but not by way of limitation, various embodiments of the present application.

[0039] FIGs. 1 A-1C: Synthesis and evaluation of degradable polyplexes. FIG. 1A: Chemical structure of the backbone-degradable copolymer containing hydrophilic (2-hydroxy ethyl methacrylate, HEMA). cationic (2-aminoethyl methacrylamide. AEMAm), and macrocyclic allylic sulfide monomer (Cycl). FIG. IB: Ester groups in the degradable copolymer backbone undergo hydrolytic degradation. FIG 1C: Degradable copolymers form apolyplex upon condensation with a GFP-encoded plasmid (pMAX-GFP) via electrostatic interactions. U-2 OS cells were transfected with the polyplexes formed with backbone-degradable polymers. GFP-positive cells were counted using an image cytometer.

[0040] FIGs. 2A-2D: Biodegradability of copolymers. FIG. 2A: Copolymerization with Cycl monomer incorporates ester-containing biodegradable residues into the polymer backbone. FIGs. 2B-2D: Size-exclusion chromatography (SEC) was used to analy ze polymers after incubation with NH4OH (FIG. 2B), HEPES buffer (FIG. 2C), and esterase (FIG. 2D).

[0041] Polymers synthesized without the macrocyclic co-monomer (black traces) show no change in molecular weight. In contrast, ester-containing copolymers (red traces) saw significant shifts in response to chemical and enzymatic challenge, while remaining stable under neutral conditions.

[0042] FIG. 3: Synthetic scheme for synthesis of backbone-degradable cationic copolymers via PET-RAFT polymerization. Macrocyclic allylic sulfide Cycl can participate in PET-Attorney Docket No. 370602-7086W01(00285)

[0043] RAFT via radical ring-opening cascade copolymerization (rROCCP) to introduce ester groups into the copolymer backbone.

[0044] FIGs. 4A-4C: Polymerization kinetics and incorporation of macrocyclic allylic sulfide monomer (Cycl) into backbone-degradable cationic copolymers via rROCCP. FIG. 4A: Copolymerization kinetics of a PET-RAFT reaction polymerizing co-monomers AEMAm (60 mol% feed ratio), HEMA (30 mol%), and Cycl (10 mol%). Monomer conversion is presented as percent of its respective feed. FIG. 4B: Cycl conversion as a function of overall monomer conversion across the kinetics study. Conversion continues to increase over the course of the reaction, indicating incorporation of the degradable ester sequence throughout the polymer chain. FIG. 4C: Comparison of degradable sequence frequency in copolymer backbone across five PET-RAFT polymerization reactions with varying Cycl feed ratio (0-10 mol%), showing degradability can be modulated by altering monomer feed.

[0045] FIGs. 5A-5C: Transfection efficiency of degradable and non-degradable polyplexes. Polyplexes were formed with pMAX_GFP at varied N / P ratios using degradable copolymers with variable macrocyclic biodegradable residues. U-2 OS cells were transfected with these polyplexes and were imaged at 48 h. FIG. 5A: The highest transfection efficiency was observed with degradable copolymers containing 7.5% macrocyclic monomer content at low' N / P ratios of 5 and 10. Transfection efficiency w as calculated by dividing the number of GFP-expressing cells with the total number of cells (obtained by Hoechst staining) and normalized to the highest average transfection efficiency obtained (Cycl = 7.5%, N / P = 5). FIG. 5B: The number of cells expressing GFP was high with degradable copolymers containing 7.5% and 10% of macrocyclic monomer at a low' N / P ratio of 5. FIG. 5C: The GFP mean intensity, or the average number of GFP molecules produced per cell by polyplex and commercial reagent PEIpro demonstrate no significant difference. Significance between degradable (feed ratio of macrocyclic monomer > 0) and non-degradable RAFT copolymers datasets at respective N / P ratios w ere determined using ANOVA with a Dunnett’s post hoc test (* p<0.05; ** p<0.01; *** p<0.001). All plots are plotted with mean ± SE at n = 3 replicates. No GFP expression was observed in untreated and pMAX_GFP only treated cells.

[0046] FIGs. 6A-6B: Cytotoxicity of degradable and non-degradable polyplexes. FIG. 6A: Cell death caused by polyplexes formed with pMAX_GFP at varied N / P ratios using degradable copolymers with variable macrocyclic biodegradable residues. Cell death (%) w as calculated as [(Y-X)*100 / Y] where Y is the total number of cells in the control wells and X is the total number of cells in treated wells obtained by Hoechst stain counting. Higher N / P ratios of 20 causes higher cell death. Commercial reagent PEIpro causes higher cell deathAttorney Docket No. 370602-7086W01(00285)

[0047] than polyplexes formed at N / P 5. FIG. 6B: Cell viability of polypi ex treated cells as measured by CCK-8 kit and normalized to the control cells. All polyplexes formulated with degradable copolymers with an N / P ratio of 5 demonstrate comparable cell viability to the control cells and higher cell viability than PEIpro. Significance between degradable (feed ratio of macrocyclic monomer > 0) and non-degradable RAFT copolymers datasets at respective N / P ratios were determined using ANOVA with a Dunnett’s post hoc test (* p<0.05; ** p<0.01; *** p<0.001). All plots are plotted with mean ± SE at n = 3 replicates. Cell viability and death in the pDNA treatment group were 100.59% ± 0.34% and 11.20% ± 2.34%, respectively.

[0048] FIG. 7: Normalized GFP cell count, cell death, and cell viability obtained for polyplexes formed with degradable copolymers containing 7.5% and 10% Cycl content at N / P 5 compared to its non-degradable analog containing 0% Cycl. Polyplexes formed with 7.5% and 10% Cycl content demonstrate higher GFP cell count while maintaining low cell death and high cell viability. Significance between degradable (feed ratio of macrocyclic monomer > 0) and non-degradable RAFT copolymers datasets at respective N / P ratios were determined using ANOVA with a Dunnett’s post hoc test (* p<0.05; ** p<0.01; *** p<0.001). All plots are plotted with mean ± SE at n = 3 replicates.

[0049] FIG. 8: General synthetic scheme for copolymerization of macrocyclic allylic sulfide monomer Cycl with comonomer (z.e., (meth)acrylates or (meth)acrylamides) via PET-RAFT polymerization. Cycl is able to participate in the PET-RAFT process via radical ring-opening cascade copolymerization (rROCCP).

[0050] FIG. 9:JH NMR in D2O of degradable cationic copolymer prepared with Cycl feed ratio of 10 mol%. Degradable unit incorporation calculated by the integration of aromatic protons of the degradable comonomer Cycl (A) and methyl protons of HEMA and AEMAm comonomers (B): % incorp. = [L / 5] / ([L / 5] + [Ib / 6]) x 100%. As shown in the figure, % incorp. = (0.912 / 17.578) x 100% = 5.2%.

[0051] FIG. 10:!H NMR in D2O of degradable cationic copolymer prepared with Cycl feed ratio of 7.5 mol%. Degradable unit incorporation calculated by the integration of aromatic protons of the degradable comonomer Cycl (A) and methyl protons of HEMA and AEMAm comonomers (B): % incorp. = [Ia / 5] / ([Ia / 5] + [Ib / 6]) x 100%. As shown in the figure, % incorp. = (0.402 / 17.068) x 100% = 2.4%.

[0052] FIG. 11:

[0053]

[0054] NMR in D2O of degradable cationic copolymer prepared with Cycl feed ratio of 5 mol%. Degradable unit incorporation calculated by the integration of aromatic protons of the degradable comonomer Cycl (A) and methyl protons of HEMA and AEMAmAttorney Docket No. 370602-7086W01(00285)

[0055] comonomers (B): % incorp. = [L / 5] / ([Ia / 5] + [Ib / 6]) x 100%. As shown in the figure, % incorp. = (0.364 / 17.030) x 100% = 2.1%.

[0056] FIG. 12:!H NMR in D2O of degradable cationic copolymer prepared with Cycl feed ratio of 2.5 mol%. Degradable unit incorporation calculated by the integration of aromatic protons of the degradable comonomer Cycl (A) and methyl protons of HEMA and AEMAm comonomers (B): % incorp. = [L / 5] / ([Ia / 5] + [Ib / 6]) x 100%. As shown in the figure, % incorp. = (0.106 / 16.772) x 100% = 0.6%.

[0057] FIG. 13: 'H NMR in D2O of non-degradable cationic copolymer prepared without Cycl in feed ratio.

[0058] FIG. 14: ’H NMR (DMSO-de) of reaction mixture for degradable cationic copolymer Pl (fAEMAm0= 0.6, HEMA0= 0.3, fcyd0= 0.1) at the start (Oh. top) and end (18h, bottom) of the reaction. Integrals are assigned to vinyl protons of unreacted HEMA (5 = 6.04 ppm and 8 = 5.66 ppm, “A”), vinyl protons of unreacted AEMAm (8 = 5.74 ppm and 8 = 5.36 ppm, “B”), vinyl protons of unreacted Cycl (8 = 6.34 ppm and 8 = 5.95 ppm, “C”), and allylic proton of unreacted Cycl (8 = 5.20, “D”).

[0059] FIG. 15:XH NMR (DMSO-de) of reaction mixture for degradable cationic copolymer P2 fAEMAm = 0.6, / H£M4° = 0.325, fcyd° = 0.075) at the start (Oh, top) and end (18h, bottom) of the reaction. Integrals are assigned to vinyl protons of unreacted HEMA (8 = 6.04 ppm and 8 = 5.66 ppm, “A’"), vinyl protons of unreacted AEMAm (8 = 5.74 ppm and 8 = 5.36 ppm, " B”). vinyl protons of unreacted Cycl (8 = 6.34 ppm and 8 = 5.95 ppm, “C”), and allylic proton of unreacted Cycl (8 = 5.20, "‘D”).

[0060] FIG. 16: ’H NMR (DMSO-de) of reaction mixture for degradable cationic copolymer P3 (fAEMAm0= 0.6, HEMA0= 0.35,fcyd° = 0.05) at the start (Oh, top) and end (18h, bottom) of the reaction. Integrals are assigned to vinyl protons of unreacted HEMA (8 = 6.04 ppm and 8 = 5.66 ppm, ‘" A”), vinyl protons of unreacted AEMAm (8 = 5.74 ppm and 8 = 5.36 ppm, “B”), vinyl protons of unreacted Cycl (8 = 6.34 ppm and 8 = 5.95 ppm, “C”), and allylic proton of unreacted Cycl (8 = 5.20, “D”).

[0061] FIG. 17: 'H NMR (DMSO-de) of reaction mixture for degradable cationic copolymer P4 jAEUAm0= 0.6, fnEMA0= 0.375, fcyd0= 0.025) at the start (Oh, top) and end (18h, bottom) of the reaction. Integrals are assigned to vinyl protons of unreacted HEMA (8 = 6.04 ppm and 8 = 5.66 ppm, “A”), vinyl protons of unreacted AEMAm (8 = 5.74 ppm and 8 = 5.36 ppm, “B"’), vinyl protons of unreacted Cycl (8 = 6.34 ppm and 8 = 5.95 ppm, “C”), and allylic proton of unreacted Cycl (8 = 5.20, “D”).Attorney Docket No. 370602-7086W01(00285)

[0062] FIG. 18: ’I I NMR (DMSO-de) of reaction mixture for non-degradable cationic copolymer P5 (fAEMAnP = 0.6, HEMA0= 0.4. fcyci0= 0) at the start (Oh, top) and end (18h, bottom) of the reaction. Integrals are assigned to vinyl protons of unreacted HEMA (5 = 6.04 ppm and 5 = 5.66 ppm, “A”) and vinyl protons of unreacted AEMAm (5 = 5.74 ppm and 5 = 5.36 ppm, “B”).

[0063] FIGs. 19A-19B: FIG. 19A: Fluorescence microscopy of U-2 OS cells transfected with polyplexes formed with degradable and non-degradable polyplexes with varied feed ratio of the degradable residue and varied N / P ratios. FIG. 19B: Images of U-2 OS control cells, cells transfected only with pMAX GFP without any polymer delivery vehicle, and cells transfected with PEIpro. GFP expression was measured using target expression analysis with a Celigo Image Cytometer (Nexcelom Bioscience). Green fluorescence channel (483 / 536) was used to image the GFP expression with an exposure time of 10 ms. Celigo software was used for the automated image analysis that counts the GFP-positive cells and the mean intensity of GFP expression in each cell. Scale bar = 1 mm. Untreated cells and those treated with only the pMAX-GFP vector (i.e., no polymer) did not demonstrate any transfection.

[0064] FIG. 20: Dynamic light scattering (DLS)-based characterization of polyplexes formed from polymer library. (Top) Summary of hydrodynamic radii for polypi ex library. Polyplexes were formed in PBS and measured in triplicate. Data are reported as mean ± SD. (Bottom) Representative histograms of each polyplex demonstrating formation of well-defined particles with relatively narrow dispersities and a lack of large aggregates.

[0065] FIG. 21: Complexation efficiency of cationic polymer library. (Top) Quantification of pDNA encapsulation efficiency as reported by a picogreen-based quantification assay. All polyplexes demonstrated high efficiency (>90%). (Bottom) Gel electrophoresis-based quantification of free pDNA after polyplex formation. No free pDNA is observed in the gel indicating near-quantitative incorporation of the pDNA at all N / P ratios tested.

[0066] FIG. 22: Backbone degradability of cationic copolymers. Polymers synthesized without the macrocyclic comonomer (P5) show no change in molecular weight after incubation withNH4OH. In contrast, ester-containing copolymers (Pl and P3) saw significant shifts in response to chemical challenge. Furthermore, the degree of fragmentation increased for the construct w ith a greater number of ester residues (Pl vs P3), indicating degradability of the cationic polymer system could be tuned by altering the feed ratio of the cyclic comonomer Cycl.

[0067] DETAILED DESCRIPTION OF THE INVENTIONAttorney Docket No. 370602-7086W01(00285)

[0068] Reference will now be made in detail to certain embodiments of the disclosed subject matter, examples of which are illustrated in part in the accompanying drawings. While the disclosed subject matter will be described in conjunction with the enumerated claims, it will be understood that the exemplified subject matter is not intended to limit the claims to the disclosed subject matter.

[0069] Throughout this document, values expressed in a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. For example, a range of “about 0.1% to about 5%"’ or “about 0.1% to 5%” should be interpreted to include not just about 0.1% to about 5%, but also the individual values (e., 1%. 2%, 3%, and 4%) and the sub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the indicated range. The statement “about X to Y” has the same meaning as “about X to about Y,” unless indicated otherwise. Likewise, the statement “about X, Y, or about Z” has the same meaning as “about X, about Y, or about Z,” unless indicated otherwise.

[0070] In this document, the terms “a,” “an / ’ or “the” are used to include one or more than one unless the context clearly dictates otherwise. The term “or” is used to refer to a nonexclusive “or” unless otherwise indicated. The statement “at least one of A and B” or “at least one of A or B” has the same meaning as “A, B, or A and B.” In addition, it is to be understood that the phraseology or terminology employed herein, and not otherwise defined, is for the purpose of description only and not of limitation. Any use of section headings is intended to aid reading of the document and is not to be interpreted as limiting; information that is relevant to a section heading may occur within or outside of that particular section. All publications, patents, and patent documents referred to in this document are incorporated by reference herein in their entirety, as though individually incorporated by reference.

[0071] In the methods described herein, the acts can be carried out in any order, except when a temporal or operational sequence is explicitly recited. Furthermore, specified acts can be carried out concurrently unless explicit claim language recites that they be carried out separately. For example, a claimed act of doing X and a claimed act of doing Y can be conducted simultaneously within a single operation, and the resulting process will fall within the literal scope of the claimed process.

[0072] Description

[0073] Gene therapy presents a promising alternative to traditional therapeutics in treatingAttorney Docket No. 370602-7086W01(00285)

[0074] hereditary as well as non-hereditary diseases including genetic disorders, neurological disorders, cardiovascular diseases, and cancer. It involves using DNA and RNA molecules to correct, modify, or silence missing genes to treat otherwise incurable diseases. However, the application of gene therapy faces challenges due to the fragile nature of these therapeutic genes, which are susceptible to degradation by serum nucleases, poor membrane permeability, low cell uptake, and instability in circulation. Hence, using gene delivery vectors is critical for delivering these therapeutic genes into the target cells and ensuring efficient transfection. These vectors are divided into two main types: viral and non-viral.

[0075] Viral vectors, despite their efficiency, pose safety risks such as immune responses and toxicity. Non-viral vectors, including cationic polymers, lipids, and nanoparticles, are preferred for their lower immunogenicity, cost-effectiveness, high loading capacity, and versatility. Among these non-viral vectors, cationic polymers are highly versatile, exhibit batch-to-batch uniformity, possess precisely controlled chemical structure, and are biologically inert. Common examples of cationic polymers for gene delivery include include polyethylenimines (PEI), poly(2-N-(dimethylaminoethyl) methacrylate) (PDMAEMA), and poly(L-lysine) (PLL). Cationic polymers can condense with negatively charged genes via electrostatic interactions to form polyelectrolyte complexes also known as polyplexes. These polyplexes are taken up by cells through endocytosis mechanisms, after which they must reach the cytosol, where they can disassemble to release their gene payloads and traffic to their intracellular site of action. For example, polyplexes formed with plasmid DNA (pDNA) must translocate to the nucleus for transcription and protein expression.

[0076] The main challenge for the clinical application of cationic polymers is cytotoxicity, arising primarily due to their high molecular weight, positive charges, and non-degradable nature. Polymers like PEI, PDMAEMA. and PLL, which contain non-degradable backbones, can persist and accumulate in the body. This chronic biocompatibility concern is further exacerbated by the need for repeated administrations with many gene therapies. While using lower molecular weight polymers can mitigate toxicity, it also decreases their ability to complex with therapeutic genes and adversely affects transfection efficiency. This trade-off complicates the design and use of polyplexes for gene delivery. Biodegradable polymers have the potential to address these challenges by degrading inside the cells thus lowering their accumulation in treated cells and reducing their overall toxicity. The degradation of biodegradable polymers in physiological environments relies on hydrolysis of the polymer backbone via breakdown of labile linkages such as esters. This process allows these polyesters to be safely eliminated from the body through excretion, improving theirAttorney Docket No. 370602-7086W01(00285)

[0077] biocompatibility and biosafety.

[0078] In addition, they can undergo hydrolysis without the need of enzymes.

[0079] Biodegradability may also enhance disassembly kinetics of the polyplexes in the cytosol enabling more efficient payload release. The first examples of backbone degradable polyesters for gene therapy utilized poly(4-hydroxy-L-proline ester) (PHP), a biodegradable polymer, and described its degradation along with the transfection efficiency. It was found that PHP degrades to half its original molecular weight in under two hours and fully degrades in three months, showing effective DNA binding and comparable transfection efficiency to PLL, and is unaffected by serum proteins. Similarly, cationic polylactides (CPLA) synthesized via ring-opening polymerization of allyl-functionalized lactide monomers followed by converting allyl functionalities to tertiary amines also demonstrated successful gene transfection with complete hydrolytic degradation within one week. Although multiple investigations have been conducted to evaluate the transfection efficiency of backbone degradable polyesters, there is a lack of evidence of their degradation kinetics.

[0080] In one aspect, the disclosure describes improvements upon previously disclosed degradable vinyl random copolymers synthesized via radical ring-opening cascade copolymerization (rROCCP) of macrocyclic allylic sulfones to investigate the degradation and transfection efficiency of novel polyesters. The ability' of this chemistry' to incorporate biodegradable ester groups directly into the polymer backbone has the potential to significantly improve biocompatibility while enhancing gene delivery- function.

[0081] In one aspect, described herein is the development of backbone-degradable cationic polymer (BDP) using Photoinduced Electron / Energy Transfer-Reversible Addition-Fragmentation Chain-Transfer (PET-RAFT) polymerization of a cationic monomer, a hydrophilic monomer, and a macrocyclic allylic sulfide (or sulfone) as the biodegradable component. A variable number of biodegradable residues were incorporated directly into the backbone of the RAFT polymer to create 4 distinct BDPs. The BDPs were then complexed ■with a model GFP-encoded plasmid to evaluate their transfection abi 1 i ty in vitro in a model mammalian cell line. The GFP-expression was measured using a high-throughput live cell imaging plate reader, which eliminates the time-consuming sample preparation required for GFP-expression analysis using flow cytometry. Finally, the cytotoxicity' induced by these BDPs yvas also investigated to identify the top performing BDPs. These results establish a proof of concept that BDPs synthesized via PET-RAFT polymerization will further broaden the BDP combinatorial library and establish a more extensive dataset for data-driven design of degradable polymeric gene vectors that demonstrate improved biocompatibility andAttorney Docket No. 370602-7086W01(00285)

[0082] transfection.

[0083] Definitions

[0084] The term “about” as used herein can allow for a degree of variability in a value or range, for example, within 10%, within 5%, or within 1% of a stated value or of a stated limit of a range, and includes the exact stated value or range.

[0085] The term “alkenyl” as used herein refers to straight and branched chain and cyclic alkyl groups as defined herein, except that at least one double bond exists between two carbon atoms. Thus, alkenyl groups have from 2 to 40 carbon atoms, or 2 to about 20 carbon atoms, or 2 to 12 carbon atoms or, in some embodiments, from 2 to 8 carbon atoms.

[0086] Examples include, but are not limited to vinyl, -CH=C=CCH2, -CH=CH(CH3). -CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), -C(CH2CH3)=CH2, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl among others.

[0087] The term “alkoxy” as used herein refers to an oxygen atom connected to an alkyl group, including a cycloalkyl group, as are defined herein. Examples of linear alkoxy groups include but are not limited to methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, and the like. Examples of branched alkoxy include but are not limited to isopropoxy, sec-butoxy, tert-butoxy, isopentyloxy, isohexyloxy, and the like. Examples of cyclic alkoxy include but are not limited to cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like. An alkoxy group can include about 1 to about 12, about 1 to about 20, or about 1 to about 40 carbon atoms bonded to the oxygen atom, and can further include double or triple bonds, and can also include heteroatoms. For example, an allyloxy group or a methoxyethoxy group is also an alkoxy group within the meaning herein, as is a methylenedioxy group in a context where two adjacent atoms of a structure are substituted therewith.

[0088] The term “alkyl” as used herein refers to straight chain and branched alkyl groups and cycloalkyl groups having from 1 to 40 carbon atoms, 1 to about 20 carbon atoms, 1 to 12 carbons or, in some embodiments, from 1 to 8 carbon atoms. Examples of straight chain alkyl groups include those with from 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl, and 2,2-dimethylpropyl groups. As used herein, the term “alkyd” encompasses n-alkyd, isoalkyl, and anteisoalkyl groups as well as other branched chain forms of alkyl. Representative substituted alkyl groups can be substituted one or more times with any of the groups listed herein, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.Attorney Docket No. 370602-7086W01(00285)

[0089] The term “alkylene” or “alkylenyl” as used herein refers to a bivalent saturated aliphatic radical (e.g., -CH2-, -CH2CH2-. and -CH2CH2CH2-, inter alia). In certain embodiments, the term may be regarded as a moiety derived from an alkene by opening of the double bond or from an alkane by removal of two hydrogen atoms from the same (e.g., -CH2-) different (e.g., -CH2CH2-) carbon atoms.

[0090] The term “alkynyl” as used herein refers to straight and branched chain alkyl groups, except that at least one triple bond exists between two carbon atoms. Thus, alkynyl groups have from 2 to 40 carbon atoms, 2 to about 20 carbon atoms, or from 2 to 12 carbons or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to –C≡CH, -C≡C(CH3), -C≡C(CH2CH3), -CH2C≡CH, -CH2C≡C(CH3), and -CH2C≡C(CH2CH3) among others.

[0091] The term “aryl” as used herein refers to cyclic aromatic hydrocarbon groups that do not contain heteroatoms in the ring. Thus aryl groups include, but are not limited to, pheny l, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl. biphenylenyl, anthracenyl, and naphthyl groups. In some embodiments, aryl groups contain about 6 to about 14 carbons in the ring portions of the groups. Aryl groups can be unsubstituted or substituted, as defined herein. Representative substituted aryl groups can be mono-substituted or substituted more than once, such as, but not limited to, a phenyl group substituted at any one or more of 2-, 3-, 4-. 5-, or 6-positions of the phenyl ring, or a naphthyl group substituted at any one or more of 2- to 8-positions thereof.

[0092] The term “biodegradable” or “degradable” as used herein refers to a material that can be broken down by either chemical or physical process (e.g, hydrolysis), which may be natural or artificial, upon interaction with the physiological environment at an implantation site or a treatment facility, and erodes or dissolves within a period of time, typically within days, weeks or months. A biodegradable or degradable material serves a temporary function, such as packaging materials, and may be subsequently broken down into monomeric or oligomeric components.

[0093] The term “cationic moiety” as used herein refers to a chemical entity that carries a positive charge or is capable of carrying a positive charge upon protonation, alkylation, or otherwise. In certain embodiments, the term “cationic moiety” refers to nitrogen-based groups that can exist in either their protonated (cationic) or neutral (unprotonated) forms, depending on the environmental pH. Examples of cationic moieties include, but are notAttorney Docket No. 370602-7086W01(00285)

[0094] limited to, amines (e.g., primary, secondary, tertiary amines, or quaternary ammonium groups). These moieties can contribute to the overall charge and interaction properties of a material, such as its solubility, binding affinity, or compatibility with negatively charged counterparts in various applications. The neutral (unprotonated) form of these groups is also encompassed within the scope of the term, recognizing their potential to become positively charged under appropriate conditions.

[0095] The term "cleax able” as used herein refers to a group or bond within a molecule, or a fragment thereof, which can be broken by a chemical reaction. In certain embodiments, the cleavable moiety is cleaved by hydrolysis (e.g, reaction with water). In certain embodiments, the cleavage may be facilitated and / or accelerated by the addition of one or more agents (e.g., acid or base). In certain embodiments, the cleavable moiety is cleaved by a substance that is naturally present or artificially introduced. In certain embodiments, cleavage may occur through enzymatic action (e.g., by hydrolases and proteases, inter alia).

[0096] The term “contacting” as used herein refers to bringing two or more materials into close enough proximity such that the two materials can physically interact.

[0097] The term “copolymer” as used herein refers to a polymer resulting from the polymerization reaction of at least two different monomers.

[0098] The term “covalent linkage” as used herein refers to the feature of two or more molecules being linked together by at least one covalent bond.

[0099] The term “cycloalkyl” as used herein refers to cyclic alkyl groups such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some embodiments, the cycloalkyl group can have 3 to about 8-12 ring members, whereas in other embodiments the number of ring carbon atoms range from 3 to 4, 5. 6, or 7. Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbomyl, adamantyl, bomyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and the like. Cycloalkyl groups also include rings that are substituted with straight or branched chain alkyl groups as defined herein. Representative substituted cycloalkyl groups can be mono-substituted or substituted more than once, such as, but not limited to, 2,2-, 2,3-, 2,4- 2,5- or 2,6-disubstituted cyclohexyl groups or mono-, di- or tri-substituted norbomyl or cycloheptyl groups, which can be substituted with, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

[0100] The term “cycloalkylene” or “cycloalkylenyl” as used herein refers to a bivalent saturated cycloalkyl radical (e.g.. A, V

[0101]

[0102] ,<\\ U, and '" O'", inter alia). In certainAttorney Docket No. 370602-7086W01(00285)

[0103] embodiments, the term may be regarded as a product of removal of two hydrogen atoms from the corresponding cycloalkane (e.g. cyclobutyl) by removal of two hydrogen atoms from the same (e.g.,<^>) different (e.g., U and "' "") carbon atoms.

[0104] The terms “halo,"’ “halogen,” or “halide” group, as used herein, by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.

[0105] The term “haloalkyl” group, as used herein, includes mono-halo alkyl groups, polyhalo alkyl groups wherein all halo atoms can be the same or different, and per-halo alkyl groups, wherein all hydrogen atoms are replaced by halogen atoms, such as fluoro. Examples of haloalkyl include trifluoromethyl. 1,1 -dichloroethyl, 1.2-di chloroethyl, l,3-dibromo-3,3-difluoropropyl, perfluorobutyl, and the like.

[0106] The term “heteroaryl” as used herein refers to aromatic ring compounds containing 5 or more ring members, of which, one or more is a heteroatom such as, but not limited to, N, O, and S; for instance, heteroaryl rings can have 5 to about 8-12 ring members. A heteroaryl group is a variety of a heterocyclyl group that possesses an aromatic electronic structure. A heteroaryl group designated as a C2-heteroaryl can be a 5-ring with two carbon atoms and three heteroatoms, a 6-ring with two carbon atoms and four heteroatoms and so forth.

[0107] Likewise a C4-heteroaryl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, and so forth. The number of carbon atoms plus the number of heteroatoms sums up to equal the total number of ring atoms. Heteroaryl groups include, but are not limited to, groups such as pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl, benzofuranyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl. benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups. Heteroaryl groups can be unsubstituted, or can be substituted with groups as is discussed herein. Representative substituted heteroaryl groups can be substituted one or more times with groups such as those listed herein.

[0108] Additional examples of aryl and heteroaryl groups include but are not limited to phenyl, biphenyl, indenyl, naphthyl (1 -naphthyl, 2-naphthyl), N-hydroxytetrazolyl, N-hydroxytriazolyl, N-hydroxyimidazolyl, anthracenyl (1-anthracenyl, 2-anthracenyl, 3-anthracenyl). thiophenyl (2 -thienyl, 3-thienyl), furyl (2-furyl, 3-furyl), indolyl. oxadiazolyl, isoxazolyl, quinazolinyl, fluorenyl, xanthenyl, isoindanyl, benzhydryl, acridinyl, thiazolyl,Attorney Docket No. 370602-7086W01(00285)

[0109] pyrrolyl (2-pyrrolyl), pyrazolyl (3-pyrazolyl), imidazolyl (1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), triazolyl (1,2,3-triazol-l-yl, l,2,3-triazol-2-yl l,2,3-triazol-4-yl. l,2,4-triazol-3-yl), oxazolyl (2-oxazolyl, 4-oxazolyL 5-oxazolyl), thiazolyl (2 -thiazolyl, 4-thiazolyl, 5-thiazolyl), pyridyl (2-pyridyl, 3-pyridyl, 4-pyridyl), pyrimidinyl (2-pyrimidinyl, 4-pyrimidinyl, 5 -pyrimidiny l, 6-pyrimidinyl), pyrazinyl, pyridazinyl (3- pyridazinyl, 4-pyridazinyl, 5 -pyridazinyl), quinolyl (2-quinolyl, 3-quinolyl, 4-quinolyl, 5-quinolyl, 6-quinolyl, 7-quinolyL 8-quinolyl), isoquinolyl (1 -isoquinolyl, 3-isoquinolyl, 4-isoquinolyl, 5-isoquinolyl, 6-isoquinolyl, 7-isoquinolyl, 8-isoquinolyl), benzo[b]furanyl (2-benzo[b]furanyl, 3-benzo[b]furanyl, 4-benzo[b] furanyl, 5-benzo[b]furanyl, 6-benzo[b] furanyl, 7-benzo[b] furanyl), 2,3-dihydro-benzo[b]furanyl (2-(2,3-dihydro-benzo[b]furanyl), 3-(2,3-dihydro-benzo[b] furanyl), 4-(2,3-dihydro-benzo[b]furanyl), 5-(2.3-dihydro-benzo[b]furanyl), 6-(2,3-dihydro-benzo[b]furanyl), 7-(2,3-dihydro-benzo[b]furanyl), benzo[b]thiophenyl (2-benzo[b]thiophenyl, 3-benzo[b]thiophenyl, 4-benzo[b]thiophenyl, 5-benzo[b]thiophenyl, 6-benzo[b]thiophenyl, 7-benzo[b]thiophenyl), 2,3-dihydro-benzo[b]thiophenyl, (2-(2,3-dihydro-benzo[b]thiophenyl), 3-(2,3-dihydro-benzo[b]thiophenyl), 4-(2,3-dihydro-benzo|bjthiophenyl). 5-(2,3-dihydro-benzo|bJthiophenyl), 6-(2,3-dihydro-benzo[b]thiophenyl), 7-(2,3-dihydro-benzo[b]thiophenyl), indolyl (1-indolyl, 2-indolyl, 3-indolyl, 4-indolyl, 5-indolyl, 6-indolyl, 7-indolyl), indazole (1-indazolyl, 3-indazolyl, 4-indazolyl, 5-indazolyl, 6-indazolyl, 7-indazolyl), benzimidazolyl (1 -benzimidazolyl, 2-benzimidazolyl, 4-benzimidazolyl, 5 -benzimidazolyl. 6-benzimidazolyl, 7-benzimidazolyl, 8-benzimidazolyl), benzoxazolyl (1-benzoxazolyl, 2-benzoxazolyl), benzothiazolyl (1-benzothiazolyl, 2-benzothiazolyl, 4-benzothiazolyl, 5 -benzothiazolyl, 6-benzothiazolyl, 7-benzothiazolyl), carbazolyl (1-carbazolyl, 2-carbazolyl, 3-carbazolyl, 4-carbazolyl), 5H-dibenz[b.f] azepine (5H-dibenz[b,f| azepin- 1-yl, 5H-dibenz[b,f|azepine-2-yl,

[0110] 5H-dibenz[b,f]azepine-3-yl, 5H-dibenz[b,f]azepine-4-yl, 5H-dibenz[b,f]azepine-5-yl), 10,11 -dihy dro-5H-dibenz[b,f] azepine ( 10, 11 -dihy dro-5H-dibenz[b,f] azepine- 1 -yl,

[0111] 10,1 l-dihydro-5H-dibenz[b,f]azepine-2-yl, 10,1 l-dihydro-5H-dibenz[b,f|azepine-3-yl, 10,1 l-dihydro-5H-dibenz[b,f]azepine-4-yl, 10,1 l-dihydro-5H-dibenz[b,f|azepine-5-yl), and the like.

[0112] The term “heteroarylalkyl” as used herein refers to alkyl groups as defined herein in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to a heteroaryl group as defined herein.

[0113] The term “heteroarylene’?or “heteroarylenyf’ as used herein refers to a bivalent heteroaryl radical (e.g., 2,4-pyridylene). In certain embodiments, the term may be regarded asAttorney Docket No. 370602-7086W01(00285)

[0114] a divalent radical formed by the removal of two hydrogen atoms from one or more rings of a heteroaryl moiety, wherein the hydrogen atoms may be removed from the same or different rings, preferably the same ring.

[0115] The term “heterocycloalkyl” as used herein refers to an aliphatic, partially unsaturated or fully saturated, 3- to 14-membered ring system, including single rings of 3 to 8 atoms and bi- and tricyclic ring systems where at least one of the carbon atoms of the ring is replaced with a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus. A heterocycloalkyl can include one to four heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein a nitrogen and sulfur heteroatom optionally can be oxidized and a nitrogen heteroatom can be optionally substituted. Representative heterocycloalkyl groups include, but are not limited, to the following exemplary groups: pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, and tetrahydrofuryl. The term heterocycloalkyl group can also be a C2 heterocycloalkyl, C2-C3 heterocycloalkyl, C2-C4 heterocycloalkyl, C2-C5 heterocycloalkyl, C2-C6 heterocycloalkyl, C2-C7 heterocycloalkyl, C2-C8 heterocycloalkyl, C2-C9 heterocycloalkyl, C2-C10 heterocycloalkyl, C2-C11 heterocycloalkyl, and the like, up to and including a C2-145heterocycloalkyl. For example, a C2heterocycloalkyl comprises a group which has two carbon atoms and at least one heteroatom, including, but not limited to, aziridiny 1, diazetidinyl, oxiranyl, thiiranyl, and the like. Alternatively, for example, a C5heterocycloalkyl comprises a group which has five carbon atoms and at least one heteroatom, including, but not limited to, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, diazepanyl, and the like. It is understood that a heterocycloalky l group may be bound either through a heteroatom in the ring, where chemically possible, or one of carbons comprising the heterocycloalkyl ring. The heterocycloalkyl group can be substituted or unsubstituted.

[0116] The term “heterocycloalkylene” or “heterocycloalkylenyl” as used herein refers to a _ H N.: HN— 1 / — \ \ bivalent saturated cycloalkyl radical (e.g.,

[0117]

[0118] V_ / , 7, W1, andN' — / N, inter alia). In certain embodiments, the term may be regarded as a product of removal of two hydrogen atoms from the corresponding heterocycloalkane (e.g., piperidine) by removal ofttwo. hyd.rogen a ttoms f from t thhe same t (e.g., ' \ / ) differen tt r (e.g. " O ^NHHan dd - --T ) carbon atom(s) and / or heteroatom(s).

[0119] The term “hydrolytic cleavage” as used herein refers to the cleavage of a bond in aAttorney Docket No. 370602-7086W01(00285)

[0120] moiety, molecule, or fragment thereof, by the addition of a water molecule across the bond (e.g, hydrolytic cleavage of an ester provides a carboxylic acid and an alcohol).

[0121] The term '‘hydrophilic moiety’’ as used herein refers to a chemical entity that increases the hydrophilicity of a polymer. Hydrophilic moieties may include polar, neutral functional groups that enhance the polymer's solubility or dispersibility in aqueous environments. Examples of hydrophilic moieties include hydrophobic functional groups and / or nonionic water-soluble polymers, such as polyethylene glycol (PEG), as well as other hydrophilic side chains. The term “hydrophilic functional group,” as used herein, refers to a functional group that is polar and neutral, contributing to the hydrophilicity of a polymer or other material. Such groups may include, but are not limited to, hydroxyl, carboxamide, ester, carbonyl, sulfoxide, and ether groups, as well as substituted derivatives thereof. Hydrophilic functional groups may be attached to a polymer backbone or incorporated into side chains to enhance the material’s interaction with aqueous environments. Examples of polymers incorporating hydrophilic functional groups include those containing hydroxyl- or ether-functionalized side chains.

[0122] The term “independently selected from” as used herein refers to referenced groups being the same, different, or a mixture thereof, unless the context clearly indicates otherwise. Thus, under this definition, the phrase “X1, X2, and X3are independently selected from noble gases” would include the scenario where, for example, X1, X2, and X3are all the same, where X1. X2. and X3are all different, where X1and X2are the same but X3is different, and other analogous permutations.

[0123] The term “moiety” as used herein means a designated region, fragment or functional group of a molecule or compound. A chemical moiety is sometimes expressed as a chemical entity (e.g. a substituent or variable) embedded or appended to a molecule, compound, or chemical formula.

[0124] The term “monomer” as used herein refers to a small molecule of low molecular weight that can be chemically bonded to other monomers to form a polymer. The term “monomer” is used throughout the specification to describe chemical compounds containing at least one polymerizable double bond (e.g., cyclic and / or acyclic double bonds).

[0125] As used herein, the term “non-covalently complexed” refers to the association of two or more entities through non-covalent interactions, such as ionic bonds, hydrogen bonds, van der Waals forces, or hydrophobic interactions, without the formation of covalent chemical bonds. In certain embodiments, the term describes a vector comprising a polymer and a nucleic acid, wherein the polymer and nucleic acid are associated through electrostatic orAttorney Docket No. 370602-7086W01(00285)

[0126] other non-covalent interactions, allowing for reversible binding and maintaining the integrity of the individual components. This type of complexation facilitates interactions while preserving the chemical structure of the nucleic acid and polymer.

[0127] The term “N / P ratio” as used herein refers to the ratio of positively charged nitrogen atoms (N) in a cationic polymer composition to the negatively charged phosphate groups (P) in a nucleic acid cargo. This ratio is used to quantify the balance of charges in a system, particularly in the context of polymer-nucleic acid complexes, such as those used in gene delivery or other bioconjugation applications. The N / P ratio provides an indication of the extent of charge neutralization and electrostatic interaction between the cationic polymer and the anionic nucleic acid. A higher N / P ratio typically suggests an excess of cationic charge, which can enhance nucleic acid binding, protect the nucleic acid from degradation, and facilitate cellular uptake. Conversely, a lower N / P ratio may result in incomplete complexation or reduced stability of the polymer-nucleic acid complex. The appropriate N / P ratio is often optimized based on the specific application, ensuring a balance between complex stability, biological activity, and minimal cytotoxicity.

[0128] As used herein, the term "pharmaceutically acceptable” refers to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively non-toxic, i.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0129] Suitable pharmaceutically acceptable acid addition salts may be prepared from an inorganic acid or from an organic acid. Examples of inorganic acids include hydrochloric, hydrobromic, hydriodic, nitric, carbonic, sulfuric (including sulfate and hydrogen sulfate), and phosphoric acids (including hydrogen phosphate and dihydrogen phosphate). Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, malonic, saccharin, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic). methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic, trifluoromethanesulfonic, 2-hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic, cyclohexylaminosulfonic, stearic, alginic, β-hydroxybutyric, salicylic, galactaric and galacturonic acid.

[0130] Suitable pharmaceutically acceptable base addition salts of compounds described herein include, for example, ammonium salts, metallic salts including alkali metal, alkalineAttorney Docket No. 370602-7086W01(00285)

[0131] earth metal and transition metal salts such as, for example, calcium, magnesium, potassium, sodium and zinc salts. Pharmaceutically acceptable base addition salts also include organic salts made from basic amines such as, for example, N, N’ -dibenzyl ethylene-diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine. All of these salts may be prepared from the corresponding compound by reacting, for example, the appropriate acid or base with the compound.

[0132] As used herein, the term "pharmaceutically acceptable carrier’ or "pharmaceutically acceptable excipient” means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound described herein within or to the patient such that it may perform its intended function. Typically, such constructs are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound(s) described herein, and not injurious to the patient. Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as com starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil. safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols, such as propylene glycol: polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. As used herein, “pharmaceutically acceptable carrier” also includes any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound(s) described herein, and are physiologically acceptable to the patient. Supplementary active compounds may also be incorporated into the compositions. The “pharmaceutically acceptable carrier” may further include a pharmaceutically acceptable salt of the compound(s) described herein. Other additional ingredients that may be included in the pharmaceutical compositions used with the methods or compounds described herein are known in the art and described, for example in Remington’s Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton,Attorney Docket No. 370602-7086W01(00285)

[0133] PA), which is incorporated herein by reference.

[0134] The term “random copolymer” as used herein refers to a polymer having two or more different types of monomers joined together in the same polymer chain wherein the different monomers may be arranged in any order.

[0135] The term “substantially” as used herein refers to a majority of, or mostly, as in at least about 50%. 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more, or 100%. The term “substantially free of’ as used herein can mean having none or having a trivial amount of, such that the amount of material present does not affect the material properties of the composition including the material, such that the composition is about 0 wt% to about 5 wt% of the material, or about 0 wt% to about 1 wt%, or about 5 wt% or less, or less than, equal to, or greater than about 4.5 wt%, 4, 3.5. 3, 2.5, 2, 1.5, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.01, or about 0.001 wt% or less. The term “substantially free of’ can mean having a trivial amount of, such that a composition is about 0 wt% to about 5 wt% of the material, or about 0 wt% to about 1 wt%, or about 5 wt% or less, or less than, equal to, or greater than about 4.5 wt%, 4. 3.5, 3, 2.5, 2, 1.5, 1, 0.9. 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.01, or about 0.001 wt% or less, or about 0 wt%.

[0136] The term “substituted” as used herein in conjunction with a molecule or an organic group as defined herein refers to the state in which one or more hydrogen atoms contained therein are replaced by one or more non-hydrogen atoms. The term “functional group” or “substituent” as used herein refers to a group that can be or is substituted onto a molecule or onto an organic group. Examples of substituents or functional groups include, but are not limited to, a halogen (e.g., F, Cl, Br, and I); an oxygen atom in groups such as hydroxy groups, alkoxy groups, aryloxy groups, aralkyloxy groups, oxo(carbonyl) groups, carboxyl groups including carboxylic acids, carboxylates, and carboxylate esters; a sulfur atom in groups such as thiol groups, alky l and aryl sulfide groups, sulfoxide groups, sulfone groups, sulfonyl groups, and sulfonamide groups; a nitrogen atom in groups such as amines, hydroxyamines, nitriles, nitro groups, N-oxides, hydrazides, azides, and enamines; and other heteroatoms in various other groups. Non-limiting examples of substituents that can be bonded to a substituted carbon (or other) atom include F, Cl, Br, I, OR, OC(O)N(R)2, CN, NO, NO2, ONO2, azido, CFs, OCF3, R, O (oxo), S (thiono), C(O), S(O), methylenedioxy, ethylenedioxy, N(R)2, SR, SOR, SO2R, SO2N(R)2, SO3R, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R. C(O)N(R)2, OC(O)N(R)2, C(S)N(R)2, (CH2)O-2N(R)C(O)R, (CH2)O-2N(R)N(R)2, N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)CON(R)2, N(R)SO2R, N(R)SO2N(R)2, N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(O)N(R)2,Attorney Docket No. 370602-7086W01(00285)

[0137] N(R)C(S)N(R)2, N(COR)COR, N(OR)R, C(=NH)N(R)2, C(O)N(OR)R, and C(=NOR)R, wherein R can be hydrogen or a carbon-based moiety; for example. R can be hydrogen, (Ci-Cioo) hydrocarbyl, alkyl, acyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl, or heteroarylalkyl; or wherein two R groups bonded to a nitrogen atom or to adjacent nitrogen atoms can together with the nitrogen atom or atoms form a heterocyclyl.

[0138] Polymer Composition(s)

[0139] In one aspect, the disclosure provides a polymer composition comprising a random copolymer of at least one hydrophilic vinyl monomer, at least one cationic vinyl monomer, and at least one cyclic monomer.

[0140] In certain embodiments, the polymer composition consists essentially of a random copolymer of at least one hydrophilic vinyl monomer, at least one cationic vinyl monomer, and at least one cyclic monomer.

[0141] In certain embodiments, the polymer composition consists of a random copolymer of at least one hydrophilic vinyl monomer, at least one cationic vinyl monomer, and at least one cyclic monomer.

[0142] In certain embodiments, the least one hydrophilic vinyl monomer comprises a vinyl monomer substituted with at least one hydrophilic moiety.

[0143] In certain embodiments, the at least one cationic vinyl monomer comprises a vinyl monomer substituted with at least one cationic moiety.

[0144] In certain embodiments, the at least one cyclic monomer is a compound of formula (la):

[0145]

[0146] wherein:

[0147] Rlaand Rlbare each independently selected from the group consisting of H, optionally substituted Ci-Ce alkyl, and optionally substituted Ce-Cio aryl;

[0148] R2aand R2bare each independently selected from the group consisting of H, optionally substituted Ci-Ce alkyl, and optionally substituted Ce-Cio aryl;

[0149] R3aand R3bare each independently H;

[0150] L1comprises at least one divalent moiety selected from the group consisting of optionally substituted C1-C12 alkylenyl, optionally substituted C1-C12 heteroalkylenyl, optionallyAttorney Docket No. 370602-7086W01(00285)

[0151] substituted C3-C8 cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted Ce-Cio arylenyl, and optionally substituted C2-C10 heteroarylenyl;

[0152] X1is selected from the group consisting of -O-*, -S-*, -S-S-*, -C(=O)O-*, -OC(=O)-*, -NHC(=O)-*, -C(=O)NH-*, -C(=S)O-*, -OC(=S)-*, -OC(=O)O-*, -NHC(=S)S-*, -SC(=S)NH-*, -NHC(=O)O-*, -OC(=O)NH-*, and a bond (absent);

[0153] X2is selected from the group consisting of -O-, -S-, -S-S-, -C(=O)O-, -OC(=O)-, -NHC(=O)-, -C(=O)NH-. -C(=S)O-, -OC(=S)-, -OC(=O)O-. -NHC(=S)S-, -SC(=S)NH-, -NHC(=O)O-, -OC(=O)NH-, and a bond;

[0154] wherein no more than one of X1and X2is a bond;

[0155] Z1is selected from the group consisting of -S- and -S(=O)2-;

[0156] * indicates a bond between X1and L1; and

[0157] ** indicates a bond between X2and L1.

[0158] In certain embodiments, R1ais H. In certain embodiments, R1ais CH3.

[0159] In certain embodiments, R1bis H. In certain embodiments, R1bis CH3.

[0160] In certain embodiments, R2ais H. In certain embodiments, R2ais optionally substituted phenyl. In certain embodiments, R2ais Ph. In certain embodiments, R2bis H. In certain embodiments, R2bis optionally substituted phenyl. In certain embodiments, R2bis Ph. In certain embodiments, one of R2aand R2bis H, and one of R2aand R2bis Ph.

[0161] In certain embodiments, X1is -C(=O)O-*. In certain embodiments, X1is -OC(=O)-*. In certain embodiments, X2is **-OC(=O)-. In certain embodiments, X2is **-C(=O)O-.

[0162] In certain embodiments, L1is -(CH2)-. In certain embodiments, L1is -(CH2)2-. In certain embodiments, L1is -(CH2)3-. In certain embodiments, L1is -(CH2)4-. In certain embodiments, L1is -(CH2)5- In certain embodiments, L1is -(CH2)6-. In certain embodiments, L1is -(CH2)7-. In certain embodiments, L1is -(CH2)8-. In certain embodiments. L1is -(CH2)9-. In certain embodiments, L1is -(CH2)10-. In certain embodiments, L1is -(CH2)11-. In certain embodiments, L1is -(CH2)12-.

[0163] In certain embodiments, Z1is -S-.

[0164] In certain embodiments, the cyclic monomer of formula (la) is:

[0165]

[0166] In certain embodiments, each hydrophilic monomer is independently a compound ofAttorney Docket No. 370602-7086W01(00285)

[0167] formula (lb):

[0168] R4aL2_R6

[0169] 4b)=<

[0170] R

[0171]

[0172] 4bR5(lb),

[0173] wherein:

[0174] R4aand R4bare each independently H;

[0175] R5is selected from the group consisting of H and Ci-Ce alkyl;

[0176] R6is selected from the group consisting of C1-C24 alkyl, C1-C24 heteroalkyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, and C2-C10 heteroaryl,

[0177] wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is substituted with at least one hydrophilic moiety, and

[0178] wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is further optionally substituted with at least one substituent selected from the group consisting of halogen, optionally substituted Ci-Ce alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroaryl;

[0179] L2comprises at least one divalent moiety selected from the group consisting of -O-, -N(RA)-, -C(=O)-, -C(=NRA)-, C(=S)-, -S(=O)-, and -S(=O)2-;

[0180] each occurrence of RAis independently selected from the group consisting of H, C(=O)RB, optionally substituted Ci-Ce alkyl, optionally substituted Ci-Ce heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C10 heteroaryl; and

[0181] each occurrence of RBis independently selected from the group consisting of optionally substituted Ci-Ce alkyl, optionally substituted Ci-Ce heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroaryl.

[0182] In certain embodiments, R5is CHs.

[0183] In certain embodiments, L2is -C(=O)O-.

[0184] In certain embodiments, each hydrophilic moiety is independently selected from the group consisting of ORA, C(=O)ORB, OC(=O)RA, OC(=O)ORB, OC(=O)SRA, OC(=O)N(RA)(RA), SRA, SC(=O)RA, SC(=O)ORB, SC(=O)N(RA)(RA), S(=O)RA, S(=O)ORB, S(=O)N(RA)(RA), S(=O)2RA, S(=O)2ORB, S(=O)N(RA)(RA), N(RA)C(=O)RA, N

[0185]

[0186] (RA)C(=O)ORB, N(RA)C(=O)SRA, N(RA)C(=O)N(RA)(RA), N(RA)C(=S)RA,Attorney Docket No. 370602-7086W01(00285)

[0187] N(RA)C(=S)ORB, N(RA)C(=S)SRA, N(RA)C(=S)N(RA)(RA), N(RA)S(=O)2RA, N(RA)S(=O)2RA, CN, NO2, halogen, and optionally substituted C2-C10 heteroaryl. In certain embodiments, at least one hydrophilic moiety is OH.

[0188] In certain embodiments, R6is C1-C24 alkyl substituted with at least one OH. In certain embodiments, R6is -(CH2)2OH.

[0189] O

[0190] In certain embodiments, the hydrophilic vinyl monomer is

[0191]

[0192] I In certain embodiments, each cationic vinyl monomer is independently a compound of formula (Ic):

[0193] R7a |_3_R9

[0194]

[0195] R7bR8(Ic),

[0196] wherein:

[0197] R7aand R7bare each independently H;

[0198] R8is selected from the group consisting of H and Ci-Ce alkyl;

[0199] R9is selected from the group consisting of C1-C24 alkyl, C1-C24 heteroalkyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, Ce-Cio aryl, and C2-C10 heteroaryl,

[0200] wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyd, ary l, or heteroaryl is substituted with at least one cationic moiety or ionizable (cationic) moiety, and

[0201] wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is further optionally substituted with at least one substituent selected from the group consisting of halogen, optionally substituted C i-Cs alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroaryl;

[0202] L3comprises at least one divalent moiety selected from the group consisting of -O-, -N(RC)-, -C(=O)-. -C(=NRC)-, C(=S)-, -S(=O)-, and -S(=O)2-;

[0203] each occurrence of RCis independently selected from the group consisting of H, C(=O)RB, optionally substituted Ci-Ce alkyl, optionally substituted Ci-Cs heteroalkyl, optionally substituted Cs-Cs cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroary 1; and each occurrence of RDis independently selected from the group consisting of optionally substituted C1-C6 alkyl, optionally substituted Ci-Ce heteroalkyl, optionally substituted C3-C8Attorney Docket No. 370602-7086W01(00285)

[0204] cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroaryl.

[0205] In certain embodiments, R8is CH3.

[0206] In certain embodiments, L3is -C(=O)NH-.

[0207] In certain embodiments, each cationic moiety is independently selected from the group consisting of -[NH3]+, -[NH2(RD)]+, -[NH(RD)2]+, -[N(RD)3]+, -[C(=NH2)NH2]+, -[C(=NHRD

[0208]

[0209] )NH2]+. -[C(=NH2)NHRD]+, -[C(=NH2)N(RD)2]+, -[C(=NHRD)NHRD]+. -[C(=NHRD)N(RD)2]+, guanidinium, pyridinium, pyrimidinium, pyrazinium, pyrrolium, imidazolium, triazolium, thiazolium, oxazolium, piperidinium, morpholinium, and pyrrolidinium. In certain embodiments, at least one cationic moiety is NH3+.

[0210] In certain embodiments, each ionizable (cationic) moiety is independently selected from the group consisting of NH2, NH(RD), N(RD)2, C(=NH)NH2, C(=NH)NHRD, -[C(=NH)N(RD)2, C(=NRD)NHRD, C(=NRD)N(RD)2, guanidinyl, pyridinyl, pyrimidinyl, pyrazinyl, pyrrolyl, imidazolyl, triazolyl, thiazolyl, piperidinyl, morpholinyl, and pyrroidinyl. In certain embodiments, the at least one ionizable (cationic) moiety is NH2.

[0211] In certain embodiments, R9is C1-C24 alkyl substituted with NH2. In certain embodiments, R9is C1-C24 alkyl substituted with NH3+. In certain embodiments, R6is -(CH2)2NH2. In certain embodiments, R6is -(CH2)2NH3+.

[0212] N

[0213] In certain embodiments, the cationic vinyl monomer is

[0214]

[0215] H

[0216] In certain embodiments, the hydrophilic vinyl monomer comprises about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35. 36. 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or about 50 mol% of the polymer composition.

[0217] In certain embodiments, the cationic vinyl monomer comprises about 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or about 80 mol% of the polymer composition.

[0218] In certain embodiments, the cyclic monomer comprises about 0.1, 0.2, 0.3, 0.4. 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2. 7.3, 7.4, 7.5, 7.6. 7.7, 7.8, 7.9, 8.0. 8.1, 8.2, 8.3, 8.4. 8.5, 8.6, 8.7, 8.8. 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4,Attorney Docket No. 370602-7086W01(00285)

[0219] 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9. 15.0. 15.1, 15.2, 15.3, 15.4, 15.5, 15.6, 15.7, 15.8, 15.9, 16.0, 16.1, 16.2, 16.3, 16.4, 16.5, 16.6, 16.7, 16.8, 16.9, 17.0, 17.1, 17.2, 17.3, 17.4, 17.5, 17.6, 17.7, 17.8, 17.9, 18.0, 18.1, 18.2, 18.3, 18.4, 18.5, 18.6, 18.7, 18.8, 18.9, 19.0, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, or about 20 mol% to about 20 mol% of the polymer composition.

[0220] In certain embodiments, the hydrophilic vinyl monomer, cationic vinyl monomer, and cyclic monomer have a molar ratio of about 30:60: 10. In certain embodiments, the hydrophilic vinyl monomer, cationic vinyl monomer, and cyclic monomer have a molar ratio of about 32.5:60:7.5. In certain embodiments, the hydrophilic vinyl monomer, cationic vinyl monomer, and cyclic monomer have a molar ratio of about 35:60:5. In certain embodiments, the hydrophilic vinyl monomer, cationic vinyl monomer, and cyclic monomer have a molar ratio of about 37.5:60:2.5. In certain embodiments, the hydrophilic vinyl monomer, cationic vinyl monomer, and cyclic monomer have a molar ratio of about 28.5:54.6:5. In certain embodiments, the hydrophilic vinyl monomer, cationic vinyl monomer, and cyclic monomer have a molar ratio of about 30.9:54.6:3.6. In certain embodiments, the hydrophilic vinyl monomer, cationic vinyl monomer, and cyclic monomer have a molar ratio of about 33.3:54.6:2.5. In certain embodiments, the hydrophilic vinyl monomer, cationic vinyl monomer, and cyclic monomer have a molar ratio of about 35.6:54.6:1.1.

[0221] In certain embodiments, each occurrence of optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkylenyl, optionally substituted heteroalkylenyl, optionally substituted cycloalkylenyl, optionally substituted heterocycloalkylenyl, optionally substituted arylenyl, and optionally substituted heteroarylenyl is independently optionally substituted with at least one substituent selected from the group consisting of Ci-Ce alkyl, Cs-Cs cycloalkyl, C2-C12 heterocycloalkyl, Ci-Ce hydroxyalkyl, halogen, CN, NO2, OR1, NIR'KR11). Ci-Ce haloalkoxy, Cs-Cs halocycloalkoxy, aryl, heteroaryl, (C1-C6 alkylenyl)C(=O)N(RI)(RII), (Ci-Ce alkylenyl)C(=O)ORI, O(C1-C3 alkylenyl)C(=O)N(RI)(RII), C(=O)RI, C(=O)ORI, OC(=O)RI, O

[0222]

[0223] C(=O)OR1, SR1, S(=O)RI, S(=O)2RI, S(=O)2RI, S(=O)2N(RI)(RII), S(=O)2NRIC(=O)NHRII, N(RI)S(=O)2RII, N(RI)C(=O)RII, and C(=O)N(RI)(RII), wherein R1and Rnare each independently selected from the group consisting of H, -C(=O)(Ci-C6 alkyl), Ci-Ce alkyl, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce heteroalkyl, C3-C8 cycloalkyl, C2-C12 heterocycloalkyl, C7-C12 aralkyl, aryl, and heteroaryl.Attorney Docket No. 370602-7086W01(00285)

[0224] In another aspect, the disclosure provides a polymer composition comprising a random copolymer of formula (II):

[0225] T1-A-T2(II),

[0226] wherein:

[0227] T1is a RAFT polymerization initiating end;

[0228] T2is a RAFT polymerization terminating end;

[0229] A comprises m units of M1, n units of M2, and o units of M3;

[0230] R3a

[0231] I

[0232] M2is R6;

[0233] R7aR8

[0234] I

[0235] M

[0236]

[0237] 3is R9;

[0238] Rlaand Rlbare each independently selected from the group consisting of H, optionally substituted Ci-Ce alkyl, and optionally substituted Cs-Cio aryl;

[0239] R2aand R2bare each independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, and optionally substituted Ce-Cio ary l;

[0240] R3aand R3bare each independently H;

[0241] R4aand R4bare each independently H;

[0242] R3is selected from the group consisting of H and Ci-Ce alkyl;

[0243] R6is selected from the group consisting of C1-C24 alkyl, C1-C24 heteroalkyl, Cs-Cs cycloalky 1, C2-C8 heterocycloalkyl, Ce-Cio ary l, and C2-C10 heteroaryl,

[0244] wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is substituted with at least one hydrophilic moiety, and

[0245] wherein the alkyl, heteroalky 1, cycloalky 1, heterocycloalkyd, aryl, or heteroary 1 is further optionally substituted with at least one substituent selected from the group consisting of halogen, optionally substituted C i-Ce alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl.Attorney Docket No. 370602-7086W01(00285)

[0246] optionally substituted C2-C8 heterocycloalkyl, optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroaryl;

[0247] R7aand R7bare each independently H;

[0248] R8is selected from the group consisting of H and Ci-Ce alkyl;

[0249] R9is selected from the group consisting of C1-C24 alky l, C1-C24 heteroalkyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, Ce-Cio aryl, and C2-C10 heteroaryl,

[0250] wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is substituted with at least one cationic moiety or ionizable (cationic) moiety7, and

[0251] wherein the alky l, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is further optionally substituted with at least one substituent selected from the group consisting of halogen, optionally substituted Ci-Ce alkyl, optionally substituted Ci-Ce heteroalkyl, optionally substituted Cs-Cs cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroaryl;

[0252] L1comprises at least one divalent moiety selected from the group consisting of optionally substituted C1-C12 alkylenyl, optionally substituted C1-C12 heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted Ce-Cio arylenyl, and optionally substituted C2-C10 heteroarylenyl;

[0253] L2comprises at least one divalent moiety selected from the group consisting of -O-, -N(RA)-, -C(=O)-, -C(=NRA)-, C(=S)-, -S(=O)-, and -S(=O)2-;

[0254] L3comprises at least one divalent moiety selected from the group consisting of -O-, -N(RC)-, -C(=O)-, -C(=NRC)-, C(=S)-, -S(=O)-, and -S(=O)2-;

[0255] X1is selected from the group consisting of -O-*, -S-*, -S-S-*, -C(=O)O-*, -OC(=O)-*. -NHC(=O)-*, -C(=O)NH-*, -C(=S)O-*, -OC(=S)-*, -OC(=O)O-*, -NHC(=S)S-*, -SC(=S)NH-*, -NHC(=O)O-*, -OC(=O)NH-*, and a bond (absent);

[0256] X2is selected from the group consisting of **-O-, **-S-, **-S-S-, **-C(=O)O-, **-OC(=O)-, **-NHC(=O)-, **-C(=O)NH-, **-C(=S)O-, **-OC(=S)-, **-OC(=O)O-, **-NHC(=S)S-, **-SC(=S)NH-, **-NHC(=O)O-, **-OC(=O)NH-, and a bond;

[0257] wherein no more than one of X1and X2is a bond;

[0258] Z2is selected from the group consisting of -S- and a bond;

[0259] each occurrence of RAis independently selected from the group consisting of H, C(=O)RB, optionally substituted C1-C6 alkyl, optionally substituted Ci-Ce heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl.Attorney Docket No. 370602-7086W01(00285)

[0260] optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroaryl:

[0261] each occurrence of RBis independently selected from the group consisting of optionally substituted Ci-Ce alkyl, optionally substituted Ci-Cs heteroalkyl, optionally substituted Cs-Cs cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroaryl;

[0262] each occurrence of RCis independently selected from the group consisting of H, C(=O)RB, optionally substituted C1-C6 alkyl, optionally substituted Ci-Cs heteroalkyl, optionally substituted Cs-Cs cycloalkyl, optionally substituted C2-C8 heterocycloalky 1, optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroaryl:

[0263] each occurrence of RDis independently selected from the group consisting of optionally substituted Ci-Ce alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C10 heteroaryl;

[0264] m, n, and o are each independently an integer ranging from 0 to 10,000; and

[0265] each divalent moiety (i.e., M1, M2, and M3) has a first terminus and a second terminus;

[0266] T1has a first terminus

[0267]

[0268] and

[0269] T2has a second terminus ( — ),

[0270] wherein T1. M1, M2, M3, and T2are arranged such a bond is formed only between first and second termini.

[0271] ,4a p7a a 3 As a non-limiting example of representative connectivity, moiety

[0272]

[0273] r6 r9depicts an exemplary bond between a first terminus of M2and second terminus of M3.

[0274] In certain embodiments, the polymer composition consists essentially of a random copolymer of formula (II).

[0275] In certain embodiments, the polymer composition consists of a random copolymer of formula (II).

[0276] In certain embodiments, R1ais H. In certain embodiments, R1ais CH3.

[0277] In certain embodiments, R1bis H. In certain embodiments, R1bis CH3.

[0278] In certain embodiments, R2ais H. In certain embodiments, R2ais optionally substituted phenyl. In certain embodiments, R2ais Ph. In certain embodiments, R2bis H. InAttorney Docket No. 370602-7086W01(00285)

[0279] certain embodiments, R2bis optionally substituted phenyl. In certain embodiments, R2bis Ph. In certain embodiments, one of R2aand R2bis H, and one of R2aand R2bis Ph.

[0280] In certain embodiments, X1is -C(=O)O-*. In certain embodiments, X1is -OC(=O)-*. In certain embodiments, X2is **-OC(=O)-. In certain embodiments, X2is **-C(=O)O-.

[0281] In certain embodiments, L1is -(CH2)-. In certain embodiments, L1is -(CH2)2-. In certain embodiments, L1is -(CH2)3-. In certain embodiments, L1is -(CH2)4-. In certain embodiments, L1is -(CH2)5- In certain embodiments, L1is -(CH2)e-. In certain embodiments, L1is -(CH2)7-. In certain embodiments, L1is -(CH2)8-. In certain embodiments, L1is -(CH2)9-. In certain embodiments, L1is -(CH2)10-. In certain embodiments, L1is -(CH2)11-. In certain embodiments, L1is -(CH2)12-.

[0282] In certain embodiments, R5is CH3.

[0283] In certain embodiments, L2is -C(=O)O-.

[0284] In certain embodiments, each hydrophilic moiety is independently selected from the group consisting of ORA, C(=O)ORB, OC(=O)RA, OC(=O)ORB, OC(=O)SRA, OC(=O)N(RA)(RA), SRA, SC(=O)RA, SC(=O)ORB, SC(=O)N(RA)(RA). S(=O)RA, S(=O)ORB, S(=O)N(RA)(RA), S(=O)2RA, S(=O)2ORB, S(=O)N(RA)(RA), N(RA)C(=O)RA, N(RA)C(=O)ORB, N(RA)C(=O)SRA, N(RA)C(=O)N(RA)(RA), N(RA)C(=S)RA, N(RA)C(=S)ORB, N(RA)C(=S)SRA, N(RA)C(=S)N(RA)(RA), N(RA)S(=O)2RA, N(RA)S(=O)2RA, CN, NO2, halogen, and optionally substituted C2-Cio heteroaryl. In certain embodiments, at least one hydrophilic moiety is OH.

[0285] In certain embodiments, R6is C1-C24 alkyl substituted with at least one OH. In certain embodiments, R6is -(CH2)2OH.

[0286] In certain embodiments, R8is CH3.

[0287] In certain embodiments, L3is -C(=O)NH-.

[0288] In certain embodiments, each cationic moiety is independently selected from the group consisting of -[NH3]’, -[NH2(RD)]+, -[NH(RD)2]+, -[N(RD)3]+, -[C(=NH2)NH2]+, -[C(=NHRD)N

[0289]

[0290] H2]+, -[C(=NH2)NHRD]+, -[C(=NH2)N(RD)2]+, -[C(=NHRD)NHRD]+, -[C(=NHRD)N(RD)2]+, guanidinium, pyridinium, pyrimidinium, pyrazinium, pyrrolium. imidazolium, triazolium, thiazolium, oxazolium, piperidinium, morpholinium, and pyrrolidinium. In certain embodiments, at least one cationic moiety is NH3+.

[0291] In certain embodiments, each ionizable (cationic) moiety is independently selected from the group consisting of NH2, NH(RD), N(RD)2, C(=NH)NH2, C(=NH)NHRD, -[C(=NH)N(RD)2. C(=NRD)NHRD, C(=NRD)N(RD)2. guanidinyl, pyridinyl, pyrimidinyl pyrazinyl, pyrrolyl, imidazolyl, triazolyl, thiazolyl, piperidinyl, morpholinyl, and pyrroidinyl.Attorney Docket No. 370602-7086W01(00285)

[0292] In certain embodiments, the at least one ionizable (cationic) moiety is NH2.

[0293] In certain embodiments, R9is C1-C24 alkyl substituted with NH2. In certain embodiments, R9is C1-C24 alkyl substituted with NH3+. In certain embodiments, R6is - (CH2)2NH2. In certain embodiments, R6is -(CH2)2NH3+.

[0294] In certain embodiments,

[0295]

[0296] M1is. In certain

[0297] embodiments, M1is In certain embodiments, M2is

[0298] 0^ 43^ O NH^

[0299]

[0300] OH in certain embodiments, M3isNH2. In certain embodiments, T1is O

[0301]

[0302] N. In certain embodiments, T2is S

[0303] In certain embodiments, each occurrence of optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkylenyl, optionally substituted heteroalkylenyl, optionally substituted cycloalkylenyl, optionally substituted heterocycloalkylenyl, optionally substituted arylenyl, and optionally substituted heteroarylenyl is independently optionally substituted with at least one substituent selected from the group consisting of Ci-Ce alkyl, Cs-Cs cycloalkyl, C2-C12 heterocycloalkyl, Ci-Ce hydroxyalkyl, halogen, CN, NO2, OR1, NiR'XR11). Ci-Ce haloalkoxy, Cs-Cs halocycloalkoxy, aryl, heteroaryl, (C1-C6 alkylenyl)C(=O)N(RI)(RII), (Ci-Ce alkylenyl)C(=O)ORI, O(C1-C3alkylenyl)C(=O)N(RI)(Rn), €(=0)^, C(=O)ORI, 00(0)^, O

[0304]

[0305] C(=O)OR1, SR1, S(=O)RI, S(=O)2R1, S(=O)2R1, S(=O)2N(R1)(R11), S(=O)2NR1C(=O)NHRH, N(RI)S(=0)2R11, N(RI)C(=O)Rn, and C(=O)N(RI)(Rn), wherein R1and Rnare each independently selected from the group consisting of H, -C(=0)(Ci-C6 alkyl), Ci-Ce alkyl, Ci- Ce alkyl, Ci-Ce haloalkyl, Ci-Ce heteroalkyl, C3-C8 cycloalkyl, C2-C12 heterocycloalkyl, C7- C12 aralkyl, aryl, and heteroaryl.Attorney Docket No. 370602-7086W01(00285)

[0306] Vector(s)

[0307] In one aspect, the disclosure provides a vector comprising the polymer composition of the disclosure and at least one nucleic acid cargo. In certain embodiments, the polymer and nucleic acid cargo are non-covalently complexed. In certain embodiments, the polymer at least partially encapsulates the at least one nucleic acid cargo. In certain embodiments, the polymer encapsulates the at least one nucleic acid cargo.

[0308] In certain embodiments, the nucleic acid cargo comprises RNA. In certain embodiments, the RNA is messenger RNA (mRNA). In certain embodiments, the RNA is small interfering RNA (siRNA). In certain embodiments, the RNA is microRNA (miRNA). In certain embodiments, the RNA is small-guide RNA (sgRNA).

[0309] In certain embodiments, the nucleic acid cargo comprises DNA. In certain embodiments, the DNA is plasmid DNA. In certain embodiments, the DNA is genomic DNA. In certain embodiments, the DNA is synthetic DNA.

[0310] In certain embodiments, the vector has aN / P ratio of about 1 / 1, 2 / 1, 3 / 1, 4 / 1, 5 / 1. 6 / 1, 7 / 1, 8 / 1, 9 / 1. 10 / 1. 11 / 1. 12 / 1, 13 / 1, 14 / 1, 15 / 1, 16 / 1, 17 / 1, 18 / 1, 19 / 1, 20 / 1, 21 / 1. 22 / 1. 23 / 1.

[0311] 24 / 1, 25 / 1, 26 / 1, 27 / 1, 28 / 1, 29 / 1, 30 / 1, 29 / 1, 28 / 1, 27 / 1, 26 / 1, 25 / 1, 24 / 1, 23 / 1, 22 / 1, 21 / 1, 20 / 1, 19 / 1, 18 / 1, 17 / 1, 16 / 1, 15 / 1, 14 / 1, 13 / 1, 12 / 1, 11 / 1, 10 / 1, 9 / 1, 8 / 1, 7 / 1, 6 / 1, 5 / 1, 4 / 1, 3 / 1, or about 2 / 1. In certain embodiments, the N / P ratio indicates a ratio of cationic charge (e g, cationic nitrogen atoms) in the polymer composition (N) and anionic charge (e.g., anionic phosphate groups) in the nucleic acid cargo (P). In certain embodiments, the vector has aN / P ratio of about 5 / 1. In certain embodiments, the vector has aN / P ratio of about 10 / 1. In certain embodiments, the vector has aN / P ratio of about 20 / 1.

[0312] Methods

[0313] In one aspect, the disclosure provides a method of delivering a nucleic acid to a target cell. In certain embodiments, the method comprises administering to a subject comprising the target cell the vector of the disclosure and / or a pharmaceutical composition thereof.

[0314] In another aspect, the disclosure provides a method for performing gene therapy in a subject. In certain embodiments, the method comprises administering to the subject the vector of the disclosure and / or a pharmaceutical composition thereof.

[0315] In another aspect, the disclosure provides a method for treating, ameliorating, and / or preventing a disease or disorder in a subject. In certain embodiments, the method comprises administering to the subject the vector of the disclosure and / or a pharmaceutical compositionAttorney Docket No. 370602-7086W01(00285)

[0316] thereof.

[0317] In another aspect, the disclosure provides a method for modulating gene expression in a target cell. In certain embodiments, the method comprises administering to a subject comprising the target cell the vector of the disclosure and / or a pharmaceutical composition thereof.

[0318] Administration / Dosage / Formulations

[0319] The regimen of administration may affect what constitutes an effective amount. The therapeutic formulations may be administered to the subject either prior to or after the onset of a disease state. Further, several divided dosages, as well as staggered dosages may be administered daily or sequentially, or the dose may be continuously infused, or may be a bolus injection. Further, the dosages of the therapeutic formulations may be proportionally increased or decreased as indicated by the exigencies of the therapeutic or prophylactic situation.

[0320] Administration of the compositions described herein to a patient, preferably a mammal, more preferably a human, may be carried out using known procedures, at dosages and for periods of time effective to treat a disease state in the patient. An effective amount of the therapeutic composition necessary' to achieve a therapeutic effect may vary' according to factors such as the state of the disease or disorder in the patient; the age, sex, and weight of the patient; and the ability of the therapeutic composition to treat a disease state in the patient. Dosage regimens may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. A non-limiting example of an effective dose range for a therapeutic composition described herein is from about 1 and 5,000 mg / kg of body weight / per day. One of ordinary skill in the art would be able to study the relevant factors and make the determination regarding the effective amount of the therapeutic composition without undue experimentation.

[0321] Actual dosage levels of the active ingredients in the pharmaceutical compositions described herein may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0322] In particular, the selected dosage level depends upon a variety of factors including the activity of the particular composition employed, the time of administration, the rate of excretion of the composition, the duration of the treatment, other drugs, compounds orAttorney Docket No. 370602-7086W01(00285)

[0323] materials used in combination with the composition, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well, known in the medical arts.

[0324] A medical doctor, e.g., physician or veterinarian, having ordinary skill in the art may readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the composition described herein employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0325] In particular embodiments, it is especially advantageous to formulate the composition in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the patients to be treated; each unit containing a predetermined qquantity of therapeutic composition calculated to produce the desired therapeutic effect in association with the required pharmaceutical vehicle. The dosage unit forms of the composition(s) described herein are dictated by and directly dependent on (a) the unique characteristics of the therapeutic composition and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding / formulating such a therapeutic composition.

[0326] In certain embodiments, the compositions described herein are formulated using one or more pharmaceutically acceptable excipients or carriers. In certain embodiments, the pharmaceutical compositions described herein comprise a therapeutically effective amount of a composition described herein and a pharmaceutically acceptable carrier.

[0327] The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity may be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms may be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it is preferable to include isotonic agents, for example, sugars, sodium chloride, or poly alcohols such as mannitol and sorbitol, in the composition. Prolonged absorption of the injectable compositions may be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate or gelatin.

[0328] In certain embodiments, the compositions described herein are administered to theAttorney Docket No. 370602-7086W01(00285)

[0329] patient in dosages that range from one to five times per day or more. In other embodiments, the compositions described herein are administered to the patient in range of dosages that include, but are not limited to, once every day, every two, days, every three days to once a week, and once every two weeks. It is readily apparent to one skilled in the art that the frequency of administration of the various combination compositions described herein varies from individual to individual depending on many factors including, but not limited to, age. disease or disorder to be treated, gender, overall health, and other factors. Thus, administration of the compositions and compositions described herein should not be construed to be limited to any particular dosage regime and the precise dosage and composition to be administered to any patient is determined by the attending physician taking all other factors about the patient into account.

[0330] The composition(s) described herein for administration may be in the range of from about 1 pg to about 10,000 mg, about 20 pg to about 9,500 mg, about 40 pg to about 9,000 mg, about 75 pg to about 8,500 mg, about 150 pg to about 7,500 mg, about 200 pg to about 7,000 mg, about 350 pg to about 6,000 mg, about 500 pg to about 5,000 mg, about 750 pg to about 4,000 mg, about 1 mg to about 3,000 mg, about 10 mg to about 2,500 mg, about 20 mg to about 2,000 mg, about 25 mg to about 1,500 mg, about 30 mg to about 1,000 mg, about 40 mg to about 900 mg, about 50 mg to about 800 mg, about 60 mg to about 750 mg, about 70 mg to about 600 mg, about 80 mg to about 500 mg, and any and all whole or partial increments therebetween.

[0331] In some embodiments, the dose of a composition described herein is from about 1 mg and about 2,500 mg. In some embodiments, a dose of a composition described herein used in compositions described herein is less than about 10,000 mg, or less than about 8,000 mg, or less than about 6,000 mg, or less than about 5.000 mg, or less than about 3.000 mg, or less than about 2,000 mg, or less than about 1,000 mg, or less than about 500 mg, or less than about 200 mg, or less than about 50 mg. Similarly, in some embodiments, a dose of a second composition as described herein is less than about 1,000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 400 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 40 mg, or less than about 30 mg, or less than about 25 mg, or less than about 20 mg, or less than about 15 mg, or less than about 10 mg, or less than about 5 mg, or less than about 2 mg, or less than about 1 mg, or less than about 0.5 mg, and any and all whole or partial increments thereof.

[0332] In various embodiments, the composition(s) described herein can be administered to aAttorney Docket No. 370602-7086W01(00285)

[0333] subject in an amount ranging from about 0.01 mg / kg to about 200 mg / kg, or about 0.5 mg / kg to about 190 mg / kg, or about 0.75 mg / kg to about 180 mg / kg, or about 1 mg / kg to about 170 mg / kg, or about 1.5 mg / kg to about 160 mg / kg, or about 2 mg / kg to about 150 mg / kg, or about 2.5 mg / kg to about 140 mg / kg, or about 3 mg / kg to about 130 mg / kg, or about 3.5 mg / kg to about 120 mg / kg, or about 4 mg / kg to about 110 mg / kg, or about 4.5 mg / kg to about 100 mg / kg, or about 5 mg / kg to about 95 mg / kg, or about 5.5 mg / kg to about 90 mg / kg, or about 6 mg / kg to about 85 mg / kg. or about 6.5 mg / kg to about 80 mg / kg, or about 7 mg / kg to about 75 mg / kg, or about 7.5 mg / kg to about 70 mg / kg, or about 8 mg / kg to about 65 mg / kg, or about 8.5 mg / kg to about 60 mg / kg, or about 9 mg / kg to about 55 mg / kg or about 9.5 mg / kg to about 50 mg / kg, or about 10 mg / kg to about 45 mg / kg.

[0334] In various embodiments, the composition(s) described herein can be administered to a subject in an amount that is less than, equal to, or greater than about 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, 1 mg / kg, 1.5 mg / kg, 2 mg / kg, 2.5 mg / kg, 3 mg / kg, 3.5 mg / kg, 4 mg / kg, 4.5 mg / kg, 5 mg / kg, 5.5 mg / kg, 6 mg / kg, 6.5 mg / kg, 7 mg / kg, 7.5 mg / kg, 8 mg / kg, 8.5 mg / kg, 9 mg / kg, 9.5 mg / kg, 10 mg / kg, 12 mg / kg, 14 mg / kg. 16 mg / kg, 18 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg, 50 mg / kg, 55 mg / kg, 60 mg / kg, 65 mg / kg, 70 mg / kg, 75 mg / kg, 80 mg / kg, 85 mg / kg, 90 mg / kg, 100 mg / kg, 105 mg / kg, 110 mg / kg, 115 mg / kg, 120 mg / kg, 125 mg / kg, 130 mg / kg, 140 mg / kg, 145 mg / kg. 150 mg / kg, 155 mg / kg, 160 mg / kg, 170 mg / kg, 175 mg / kg. 180 mg / kg, 185 mg / kg. 190 mg / kg. 195 mg / kg, or 200 mg / kg.

[0335] In certain embodiments, a composition as described herein is a packaged pharmaceutical composition comprising a container holding a therapeutically effective amount of a composition described herein, alone or in combination with a second pharmaceutical agent; and instructions for using the composition to treat, prevent, or reduce one or more symptoms of a disease state or disorder in a patient.

[0336] Formulations may be employed in admixtures with conventional excipients, i.e., pharmaceutically acceptable organic or inorganic carrier substances suitable for oral, parenteral, nasal, intravenous, subcutaneous, enteral, or any other suitable mode of administration, known to the art. The pharmaceutical preparations may be sterilized and if desired mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure buffers, coloring, flavoring and / or aromatic substances and the like. They may also be combined where desired with other active agents, e.g., other analgesic agents.

[0337] Routes of administration of any of the compositions described herein include oral,Attorney Docket No. 370602-7086W01(00285)

[0338] nasal, rectal, intravaginal, parenteral, buccal, sublingual or topical. The compositions for use in the compositions described herein can be formulated for administration by any suitable route, such as for oral or parenteral, for example, transdermal, transmucosal (e.g, sublingual, lingual, (trans)buccal, (trans)urethral, vaginal (e.g, trans- and perivaginally), (intra)nasal and (trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastrical, intrathecal, subcutaneous, intramuscular, intradermal, intra-arterial, intravenous, intrabronchial, inhalation, and topical administration.

[0339] Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gel caps, troches, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs, suppositories, liquid sprays for nasal or oral administration, dry powder or aerosolized formulations for inhalation, compositions and formulations for intravesical administration and the like. It should be understood that the formulations and compositions described herein are not limited to the particular formulations and compositions that are described herein.

[0340] Oral Administration

[0341] For oral application, particularly suitable are tablets, dragees, liquids, drops, suppositories, or capsules, caplets and gelcaps. The compositions intended for oral use may¬ be prepared according to any method known in the art and such compositions may contain one or more agents selected from the group consisting of inert, non-toxic pharmaceutically excipients that are suitable for the manufacture of tablets. Such excipients include, for example an inert diluent such as lactose: granulating and disintegrating agents such as cornstarch; binding agents such as starch; and lubricating agents such as magnesium stearate. The tablets may be uncoated or they may be coated by known techniques for elegance or to delay the release of the active ingredients. Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed wi th an inert diluent.

[0342] For oral administration, the composition(s) described herein can be in the form of tablets or capsules prepared by conventional means with pharmaceutically acceptable excipients such as binding agents (e.g, polyvinylpyrrolidone, hydroxypropylcellulose or hydroxypropyl methylcellulose); fillers (e.g, cornstarch, lactose, microcrystalline cellulose or calcium phosphate); lubricants (e.g, magnesium stearate, talc, or silica); disintegrates (e.g., sodium starch gly collate); or wetting agents (e.g, sodium lauryl sulphate). If desired, the tablets may be coated using suitable methods and coating materials such as OPADRY™ filmAttorney Docket No. 370602-7086W01(00285)

[0343] coating systems available from Colorcon, West Point, Pa. (e.g., OPADRY™ OY Type, OYC Type, Organic Enteric OY-P Type, Aqueous Enteric OY-A Type, OY-PM Type and OPADRY™ White, 32K18400). Liquid preparation for oral administration may be in the form of solutions, syrups or suspensions. The liquid preparations may be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, methyl cellulose or hydrogenated edible fats); emulsifying agent (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters or ethyl alcohol); and preservatives (e g, methyl or propyl p-hydroxy benzoates or sorbic acid).

[0344] Compositions as described herein can be prepared, packaged, or sold in a formulation suitable for oral or buccal administration. A tablet that includes a composition as described herein can. for example, be made by compressing or molding the active ingredient, optionally with one or more additional ingredients. Compressed tablets may be prepared by compressing, in a suitable device, the active ingredient in a free-flowing form such as a powder or granular preparation, optionally mixed with one or more of a binder, a lubricant, an excipient, a surface active agent, and a dispersing agent. Molded tablets may be made by molding, in a suitable device, a mixture of the active ingredient, a pharmaceutically acceptable carrier, and at least sufficient liquid to moisten the mixture. Pharmaceutically acceptable excipients used in the manufacture of tablets include, but are not limited to, inert diluents, granulating and disintegrating agents, dispersing agents, surface-active agents, disintegrating agents, binding agents, and lubricating agents.

[0345] Suitable dispersing agents include, but are not limited to, potato starch, sodium starch gly collate, pol oxamer 407, or poloxamer 188. One or more dispersing agents can each be individually present in the composition in an amount of about 0.01% w / w to about 90% w / w relative to weight of the dosage form. One or more dispersing agents can each be individually present in the composition in an amount of at least, greater than, or less than about 0.01%, 0.05%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or 90% w / w relative to weight of the dosage form.

[0346] Surface-active agents (surfactants) include cationic, anionic, or non-ionic surfactants, or combinations thereof. Suitable surfactants include, but are not limited to, behentrimonium chloride, benzalkonium chloride, benzethonium chloride, benzododecinium bromide, carbethopendecinium bromide, cetalkonium chloride, cetrimonium bromide, cetrimonium chloride, cetvlpyridine chloride, didecyldimethylammonium chloride, dimethyldioctadecylammonium bromide, dimethyldioctadecylammonium chloride, domiphenAttorney Docket No. 370602-7086W01(00285)

[0347] bromide, lauryl methyl gluceth-10 hydroxypropyl dimonium chloride, tetramethylammonium hydroxide, thonzonium bromide, stearalkonium chloride, octenidine dihydrochloride, olaflur, N-oleyl-l,3-propanediamine, 2-acrylamido-2 -methylpropane sulfonic acid, alkylbenzene sulfonates, ammonium lauryl sulfate, ammonium perfluorononanoate, docusate, disodium cocoamphodiacetate, magnesium laureth sulfate, perfluorobutanesulfonic acid, perfluorononanoic acid, perfluorooctanesulfonic acid, perfluorooctanoic acid, potassium lauryl sulfate, sodium alkyl sulfate, sodium dodecyl sulfate, sodium laurate, sodium laureth sulfate, sodium lauroyl sarcosinate, sodium myreth sulfate, sodium nonanoyloxybenzenesulfonate, sodium pareth sulfate, sodium stearate, sodium sulfosuccinate esters, cetomacrogol 1000, cetosteaiyl alcohol, cetyl alcohol, cocamide diethanolamine, cocamide monoethanolamine, decyl glucoside, decyl polyglucose, glycerol monostearate, octylphenoxypolyethoxyethanol CA-630, isoceteth-20, lauryl glucoside, octylphenoxypoly ethoxyethanol P-40, Nonoxynol-9, Nonoxynols, nonyl phenoxypolyethoxylethanol (NP-40), octaethylene glycol monododecyl ether, N-octyl beta-D-thioglucopyranoside, octyl glucoside, oleyl alcohol, PEG- 10 sunflower glycerides, pentaethylene glycol monododecyl ether, polidocanol, poloxamer, poloxamer 407, polyethoxylated tallow amine, polyglycerol polyricinoleate, polysorbate, polysorbate 20, polysorbate 80, sorbitan, sorbitan monolaurate, sorbitan monostearate, sorbitan tristearate, stearyl alcohol, surfactin, Triton X-100, and Tween 80. One or more surfactants can each be individually present in the composition in an amount of about 0.01% w / w to about 90% w / w relative to weight of the dosage form. One or more surfactants can each be individually present in the composition in an amount of at least, greater than, or less than about 0.01%, 0.05%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%. 60%. 65%. 70%. 75%. 80%. 85%. or 90% w / w relative to weight of the dosage form.

[0348] Suitable diluents include, but are not limited to, calcium carbonate, magnesium carbonate, magnesium oxide, sodium carbonate, lactose, microcrystalline cellulose, calcium phosphate, calcium hydrogen phosphate, and sodium phosphate, Cellactose ® 80 (75 % a-lactose monohydrate and 25 % cellulose powder), mannitol, pre-gelatinized starch, starch, sucrose, sodium chloride, talc, anhydrous lactose, and granulated lactose. One or more diluents can each be individually present in the composition in an amount of about 0.01% w / w to about 90% w / w relative to weight of the dosage form. One or more diluents can each be individually present in the composition in an amount of at least, greater than, or less thanAttorney Docket No. 370602-7086W01(00285)

[0349] about 0.01%, 0.05%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%. 45%. 50%. 55%. 60%. 65%. 70%. 75%. 80%. 85%. or 90% w / w relative to weight of the dosage form.

[0350] Suitable granulating and disintegrating agents include, but are not limited to, sucrose, copovidone, com starch, microcrystalline cellulose, methyl cellulose, sodium starch gly collate, pregelatinized starch, povidone, sodium carboxy methyl cellulose, sodium alginate, citric acid, croscarmellose sodium, cellulose, carboxymethylcellulose calcium, colloidal silicone dioxide, crosspovidone and alginic acid. One or more granulating or disintegrating agents can each be individually present in the composition in an amount of about 0.01% w / w to about 90% w / w relative to weight of the dosage form. One or more granulating or disintegrating agents can each be individually present in the composition in an amount of at least, greater than, or less than about 0.01%, 0.05%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or 90% w / w relative to weight of the dosage form.

[0351] Suitable binding agents include, but are not limited to, gelatin, acacia, pre-gelatinized maize starch, polyvinylpyrrolidone, anhydrous lactose, lactose monohydrate, hydroxypropyl methylcellulose, methylcellulose, povidone, polyacrylamides, sucrose, dextrose, maltose, gelatin, polyethylene glycol. One or more binding agents can each be individually present in the composition in an amount of about 0.01% w / w to about 90% w / w relative to weight of the dosage form. One or more binding agents can each be individually present in the composition in an amount of at least, greater than, or less than about 0.01 %, 0.05%, 0.1 %, 0.5%, 1 %, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or 90% w / w relative to weight of the dosage form.

[0352] Suitable lubricating agents include, but are not limited to, magnesium stearate, calcium stearate, hydrogenated castor oil, glyceryl monostearate, glyceryl behenate, mineral oil, polyethylene glycol, poloxamer 407, poloxamer 188, sodium laureth sulfate, sodium benzoate, stearic acid, sodium stearyl fumarate, silica, and talc. One or more lubricating agents can each be individually present in the composition in an amount of about 0.01% w / w to about 90% w / w relative to weight of the dosage form. One or more lubricating agents can each be individually present in the composition in an amount of at least, greater than, or less than about 0.01%, 0.05%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or 90% w / w relative to weight of the dosage form.

[0353] Tablets can be non-coated or they may be coated using known methods to achieveAttorney Docket No. 370602-7086W01(00285)

[0354] delayed disintegration in the gastrointestinal tract of a subject, thereby providing sustained release and absorption of the active ingredient. By way of example, a material such as glyceryl monostearate or glyceryl distearate may be used to coat tablets. Further by way of example, tablets may be coated using methods described in U. S. Patent Nos. 4,256,108; 4,160,452; and 4,265,874 to form osmotically controlled release tablets. Tablets may further comprise a sweetening agent, a flavoring agent, a coloring agent, a preservative, or some combination of these in order to provide for pharmaceutically elegant and palatable preparation.

[0355] Tablets can also be enterically coated such that the coating begins to dissolve at a certain pH, such as at about pH 5.0 to about pH 7.5, thereby releasing a composition as described herein. The coating can contain, for example. EUDRAGIT ® L, S. FS. and / or E polymers with acidic or alkaline groups to allow release of a composition as described herein in a particular location, including in any desired section(s) of the intestine. The coating can also contain, for example, EUDRAGIT ® RL and / or RS polymers with cationic or neutral groups to allow for time controlled release of a composition as described herein by pH-independent swelling.

[0356] Parenteral Administration

[0357] For parenteral administration, the compositions as described herein may be formulated for injection or infusion, for example, intravenous, intramuscular or subcutaneous injection or infusion, or for administration in a bolus dose and / or continuous infusion.

[0358] Suspensions, solutions or emulsions in an oily or aqueous vehicle, optionally containing other formulatory agents such as suspending, stabilizing and / or dispersing agents may be used.

[0359] Sterile injectable forms of the compositions described herein may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol.

[0360] Among the acceptable vehicles and solvents that may be employed are water, Ringer’s solution and isotonic sodium chloride solution. Sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or di -glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically acceptable oils, such as olive oil or castor oil. especially in their polyoxy ethylated versions. These oil solutions or suspensions may also contain a long-chainAttorney Docket No. 370602-7086W01(00285)

[0361] alcohol diluent or dispersant, such as such as lauryl, stearyl, or oleyl alcohols, or similar alcohol.

[0362] Additional Administration Forms

[0363] Additional dosage forms suitable for use with the composition(s) and compositions described herein include dosage forms as described in U. S. Patents Nos. 6,340,475;

[0364] 6,488,962; 6,451,808; 5,972,389; 5,582,837; and 5,007,790. Additional dosage forms suitable for use with the composition(s) and compositions described herein also include dosage forms as described in U. S. Patent Applications Nos. 20030147952; 20030104062; 20030104053; 20030044466; 20030039688; and 20020051820. Additional dosage forms suitable for use with the compound(s) and compositions described herein also include dosage forms as described in PCT Applications Nos. WO 03 / 35041; WO 03 / 35040; WO 03 / 35029; WO 03 / 35177; WO 03 / 35039; WO 02 / 96404; WO 02 / 32416; WO 01 / 97783; WO 01 / 56544; WO 01 / 32217; WO 98 / 55107; WO 98 / 11879; WO 97 / 47285; WO 93 / 18755; and WO 90 / 11757.

[0365] Dosing

[0366] The therapeutically effective amount or dose of a composition described herein depends on the age, sex and weight of the patient, the current medical condition of the patient and the progression of a disease state in the patient being treated. The skilled artisan is able to determine appropriate dosages depending on these and other factors.

[0367] A suitable dose of a composition described herein can be in the range of from about 0.01 mg to about 5,000 mg per day, such as from about 0.1 mg to about 1,000 mg, for example, from about 1 mg to about 500 mg, such as about 5 mg to about 250 mg per day. The dose may be administered in a single dosage or in multiple dosages, for example from 1 to 4 or more times per day. When multiple dosages are used, the amount of each dosage may be the same or different. For example, a dose of 1 mg per day may be administered as two 0.5 mg doses, with about a 12-hour interval between doses.

[0368] It is understood that the amount of composition dosed per day may be administered, in non-limiting examples, every day, every other day, every 2 days, every’ 3 days, every 4 days, or every 5 days. For example, with every other day administration, a 5 mg per day dose may be initiated on Monday with a first subsequent 5 mg per day dose administered on Wednesday, a second subsequent 5 mg per day dose administered on Friday, and so on.

[0369] In the case wherein the patient’s status does improve, upon the doctor’s discretion the administration of the composition(s) described herein is optionally given continuously; alternatively, the dose of drug being administered is temporarily reduced or temporarilyAttorney Docket No. 370602-7086W01(00285)

[0370] suspended for a certain length of time (i.e., a ‘‘drug holiday'’). The length of the drug holiday optionally varies between 2 days and 1 year, including by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days. The dose reduction during a drug holiday includes from 10%-100%, including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%. 45%. 50%. 55%. 60%. 65%. 70%. 75%. 80%. 85%. 90%. 95%. or 100%.

[0371] Once improvement of the patient’s conditions has occurred, a maintenance dose is administered if necessary. Subsequently, the dosage or the frequency of administration, or both, is reduced to a level at which the improved disease is retained. In certain embodiments, patients require intermittent treatment on a long-term basis upon any recurrence of symptoms and / or infection.

[0372] The compositions described herein can be formulated in unit dosage form. The term “unit dosage form” refers to physically discrete units suitable as unitary dosage for patients undergoing treatment, with each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, optionally in association with a suitable pharmaceutical carrier. The unit dosage form may be for a single daily dose or one of multiple daily doses (e g., about 1 to 4 or more times per day). When multiple daily doses are used, the unit dosage form may be the same or different for each dose.

[0373] Toxicity and therapeutic efficacy of such therapeutic regimens are optionally determined in cell cultures or experimental animals, including, but not limited to, the determination of the LD50(the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between the toxic and therapeutic effects is the therapeutic index, which is expressed as the ratio between LD50 and ED50. The data obtained from cell culture assays and animal studies are optionally used in formulating a range of dosage for use in human. The dosage of such compositions lies preferably within a range of circulating concentrations that include the ED50 with minimal toxicity. The dosage optionally varies within this range depending upon the dosage form employed and the route of administration utilized.

[0374] EXAMPLES

[0375] Various embodiments of the present application can be better understood by reference to the following Examples which are offered by way of illustration. The scope of the present application is not limited to the Examples given herein.Attorney Docket No. 370602-7086W01(00285)

[0376] Materials and Methods

[0377] Polymer synthesis and characterization - General procedure

[0378] Polymers were synthesized via PET-RAFT using established reaction conditions. In brief, stabilized monomers were first de-inhibited before use by passing over MEHQ inhibitor removal resin. Stock solutions of chain transfer agent (CTA, 50 mM), photoinitiator tris(2-phenylpyridinato-C2,N)iridium(III) (fac-Ir(ppy)3, 1 mM), and requisite monomers were prepared in DMSO. These stocks were then used to prepare the reaction solutions.

[0379] Photopolymerization was then carried out under 450 nm LED light at room temperature to yield the polymer product. Monomer conversions were determined using ’H-NMR spectroscopy (Bruker Avance Neo 500 MHz) of crude product with mesitylene as an internal standard. Crude products were optionally purified by precipitation into hexane: acetone (10: 1) mixture three times and dried under vacuum. Monomer content of purified polymers was analyzed by 'H-NMR in D2O. A typical procedure for PET-RAFT copolymerization of each polymer are as follows:

[0380] poly(DMA)

[0381] Dimethylacrylamide (DMA) was co-polymerized with Cycl using 2-(2-carboxyethylsulfanylthiocarbonylsulfanyl)propionic acid as CTA. The reaction was carried out a final concentration of 2 M total monomer, 4 mM CTA, and 50 pM ac-Ir(ppy)3, with 2.5 mol% Cycl in the monomer feed. The reaction mixture was irradiated for 6h to yield the polymer product. Non-degradable poly(DMA) control was prepared in parallel by analogous reaction conditions, with the exception of excluding Cycl from the reaction.

[0382] Cationic copolymers

[0383] 2-aminoethyl methacrylamide (AEMAm) and 2-hydroxy ethyl methacrylate (HEMA) were co-polymerized with Cycl using 4-cyano-4-(((ethylthio)carbonothioyl)thio)pentanoic acid as CTA. Reactions were carried out at a final concentration of 1 M total monomer, 10 mM CTA, and 200 pM / oc'-Ir(ppy)3. AEMAm feed ratio was fixed at 60 mol% for all reactions, with Cycl feed ratio varying from 0-10 mol%, and HEMA filling the balance. The reaction mixture was irradiated for 18 h to yield the polymer product.

[0384] Polymer degradation - Chemical degradation

[0385] To assess degradation of polymers in response to chemical challenge, polymer reaction mixtures were diluted 100-fold into 50 mM NH4OH and the solution was incubated for 30 mins at 37°C. For DMA copolymers, the solution was then diluted 2: 1 in dimethylformamide (DMF) and molecular weight of the polymer products were thenAttorney Docket No. 370602-7086W01(00285)

[0386] analyzed via size exclusion chromatography (SEC) using an Agilent 1200 Series system with on-line UV and RI (Agilent 1260 Series) detectors. The system was equipped with two Agilent PLgel 5 pm columns in series (103and 104A, 300 x 7.5 mm). DMF supplemented with 50 mM LiBr was used as the mobile phase. Molecular weight data (Mn, Mw, and D) were determined using a series of PMMA standards of known molecular weight (Agilent EasyVial PMMA Calibration Kit) based on the respective RI chromatographs. Cationic copolymers were analyzed using an Agilent NOVEMA Max column (103A, 8 x 300 mm) with a mobile phase consisting of 0.3% formic acid + 0.1M NaCl (pH 2.5), supplemented with 0.02 wt% NaNs. Molecular weight data were determined using a series of PEG standards of known molecular weight (Agilent EasyVial PEG Calibration Kit) based on the respective RI chromatographs.

[0387] Enzymatic degradation.

[0388] Enzymatic degradation was carried out using porcine liver esterase (PLE) as a model enzyme. Fresh solutions of PLE (Sigma Aldrich) were prepared from lyophilized powder at 20 U / mL in 10 mM HEPES buffer (pH 7.0). Prior to use, activity of esterase solution was confirmed via chromogenic assay, with 4-nitrophenyl butyrate serving as substrate. Polymer reaction mixtures were then diluted 100-fold into the enzyme solution and incubated for 24 or 48h at 37°C using a shaker-heater set to 400 RPM. At the appropriate time point, the solution was removed from the heater-shaker and diluted 2: 1 in DMF to precipitate the enzyme. The solution was then centrifuged (15k RCF, 5 min. RT) to pellet protein precipitate, and the supernatant was analyzed via SEC on the same instrumentation described above. A corresponding 0 h time point was prepared by adding polymer to the enzyme solution and immediately precipitating the enzy me. Hydrolytic degradation of the polymer in the absence of esterase was assessed using an analogous procedure.

[0389] Polyplex formation

[0390] A shuttle vector, pMAX_GFP was obtained from Addgene (plasmid# 177825). Gene sequences were verified by whole plasmid sequencing (Azenta). Plasmids were transformed into DH5a competent E. coli (New England Biolabs). Colonies picked from fresh plates were grown for 12 hours at 37°C in 5 mL LB while shaking at 250 rpm. The vectors used contained a kanamycin resistance gene; kanamycin was used at concentrations of 50 pg / mL in cultures. pDNA was extracted using a Monarch® Plasmid DNA miniprep kit (New England Biolabs) using the manufacturer’s protocol. The extracted pDNA was stored at -20°C in aliquots in an elution buffer. The DNA concentration in each aliquot was measured based on its absorbance at 280 nm using a Nanodrop spectrophotometer (ThermoFisher). OneAttorney Docket No. 370602-7086W01(00285)

[0391] aliquot of pMAX_GFP was then diluted in sterile filtered DNase-free water to a concentration of 20 ng / pL. The copolymers were also diluted in sterile filtered DNase-free water to the desired concentration and mixed in equal volumes with pMAX_GFP to obtain the desired N / P ratios of 5, 10, and 20. N / P ratio is the stoichiometric ratio between the protonable nitrogen (N) in the copolymer and the anionic phosphate groups (P) present in the pMAX_GFP. These ratios were chosen based on the previously reported recommendations for transfection with cationic RAFT copolymers.43The polyplex was then incubated for 45 minutes at room temperature and then mixed with two parts volume of serum-free DMEM and incubated for an additional 45 minutes at room temperature. Polyplexes were also prepared with PEIpro (Polyplus) as the commercial transfecting agent following the manufacturer's protocol. Briefly, equal volumes of pMAX GFP (40 ng / pL) and PEIpro solution (80 pL / mL) were mixed in Opti-MEM reduced serum medium (ThermoFisher Scientific) and incubated at room temperature for 45 minutes.

[0392] Cellular Assays

[0393] The U-2 OS cell line was used to assess the transfection efficiency where cells were maintained in DMEM supplemented with 10% FBS at 37°C and 5% CO2 in 75cm2cell culture flasks. For all transfection assays, cells were seeded at 6000 cells / well at 200 pL / well in a tissue culture-treated 96-well plate and incubated at 37 °C and 5% CO2 for 24 h before transfection. The transfection protocol was adapted from Kumar, et. al. Media was aspirated after 24 h and a 200 ng / well plasmid loading was employed where cells were incubated with 60 pL of the polyplex-serum-free-DMEM solution for 4 h at 37°C and 5% CO2. After 4 h, 200 pL DMEM with 10% FBS was added to all the wells. For transfection with PEIpro, 24 h after seeding, media was aspirated, and cells were supplemented with growth medium. After 4 h, 10 pL of PEIpro-pMAX GFP polyplex was added to the growth medium-supplemented cells. Media was aspirated from all the wells after 24 h, supplementing the cells with 200 pL of fresh growth medium. The GFP expression, cell counts, and cell viability were evaluated at 48h after transfection. All treatments were performed in triplicate.

[0394] Cell viability was performed first using a CCK-8 assay (Dojindo) according to the manufacturer’s protocol. At 44 h after transfection, 20 pL of 2% solution of CCK-8 was added to the cells and incubated for a total of 4 h at 37°C and 5% CO2. Absorbance values were obtained every hour at 450 nm using a SpectraMax UV-Vis plate reader. Blank values obtained in empty wells containing only the media and CCK-8 solution were subtracted from all measurements. Absorbance values were normalized to the control cells (cells treated withAttorney Docket No. 370602-7086W01(00285)

[0395] 60 pL of DNase-free water and serum-free DMEM solution for 4h) to determine the cell viability.

[0396] GFP expression was then immediately measured using target expression analysis with a Celigo Image Cytometer (Nexcelom Bioscience). Green fluorescence channel (483 / 536 nm) was used to image the GFP expression with an exposure time of 10 ms. Celigo software was used for the automated image analysis that counts the GFP-positive cells and the mean intensity of GFP expression in each cell. After GFP expression analysis, the cells were then stained with Hoechst 33342 (ThermoFisher Scientific), a widely used dye for live cell imaging that stains the nucleus blue. The GFP expression and the Hoechst stain were not imaged simultaneously to reduce the interference caused by the overlap of the green and blue channels. Hoechst staining was carried out according to the manufacturer's protocol to determine the total number of cells in each well. Briefly, media was aspirated from all the wells and stained with 30 pL of the Hoechst solution (prepared by diluting 1:2000 in D-Phosphate Buffered Saline (D-PBS)) for 10 minutes. After 10 minutes, the cells were washed three times with 100 pL PBS and were imaged in PBS as well. Target expression analysis was again performed using the Celigo Image Cytometer with a blue (377 / 447 nm) channel with an exposure time of 300 ms. Celigo software was used for the automated image analysis that counts the Hoechst-stained cells.

[0397] Statistical Analysis

[0398] All data are reported as mean ± standard error. The number of replicates per experimental and control group is three as also described in the related Methods subsections and the figure captions. Statistical analysis of all data was processed using the Origin software package (OriginPro, USA). Statistical significance between degradable and non-degradable RAFT copolymers datasets at respective N / P ratios was determined using ANOVA with a Dunnett’s post hoc test. Data was normalized to the highest observed value for plotting transfection efficiency and GFP cell count. For cell viability, data was normalized to the control group of cells.

[0399] pDNA Complexation Efficiency Assay

[0400] Stock solutions of pDNA were prepared in sterile-filtered DNAase-free PBS (pH 7.4) at a concentration of 10 ng / pL. Stock solutions of the cationic copolymers were also prepared in the same buffer to the desired concentrations and mixed 1: 1 with the pDNA to obtain the desired N / P ratios of 5, 10, and 20 at a final concentration of 5 ng / pL pDNA. Polyplexes w ere allowed to incubate for 45 minutes at room temperature prior to measurement. Free pDNA remaining after complexation was then quantified using the Quant-iT PicoGreen dsDNAAttorney Docket No. 370602-7086W01(00285)

[0401] Assay Reagent (Thermo Fisher Scientific). The assay was then preformed according the manufacturer specifications. In brief, the Picogreen reagent was first diluted into sterile filtered DNAase-free PBS (pH 7.4). The polyplex solution was then diluted 5x in the Picogreen stock solution to obtain a final pDNA concentration of 1 ng / pL. Concentration of free pDNA was then quantified using a Spectramax M3 platereader (Ex 480 nm / Em 525 nm) by comparing against a standard curve of known pDNA concentrations. Complexation efficiency was then calculated as (Total pDNA - Free pDNA) / Total pDNA.

[0402] Gel Electrophoresis

[0403] Gel electrophoresis was performed using 1 wt% agarose gels prepared in lx Tris-Acetate-EDTA (TAE) buffer (pH 7.4). Polyplexes were prepared as described above for the Picogreen assay. Two pL of SDS-free DNAgel loading dye (Thermo Fisher Scientific) was added to 10 pL each polyplex sample prior to loading the sample in the gel and electrophoresing for 35 min at 120 V in TAE running buffer. The gel was then imaged using an Azure Biosy stems 600 Imager.

[0404] DLS Characterization

[0405] Stock solutions of pDNA were prepared in sterile-filtered DNAase-free PBS (pH 7.4) at a concentration of 80 ng / pL. Stock solutions of the cationic copolymers were also prepared in the same buffer to the desired concentration and mixed 1: 1 with the pDNA to obtain the desired N / P ratios of 5, 10. and 20 at a final concentration of 40 ng / pL pDNA. Polyplexes were allowed to incubate for 45 minutes at room temperature prior to measurement. DLS measurements were performed on a DynaPro Plate Reader III (Wyatt Technologies) and analyzed using the accompanying Dynamics 7.10 software package. For each sample, five repeat acquisitions with five second acquisition time were collected at 25 °C.

[0406] Example 1: Design and Synthesis of Exemplary Biodegradable Polymers

[0407] The macrocyclic allylic sulfide Cycl was utilized as a comonomer to directly introduce ester groups into the polymer backbone during the polymerization process (FIG. 8). Poly(DMA) was selected as an initial water-soluble polymer system to investigate hydrolytic degradability of these esters. Towards this end, a copolymer of DMA and Cyc l (2.5 mol% feed ratio) was synthesized via PET-RAFT polymerization. A DMA homopolymer was also synthesized using identical reaction conditions to create a control polymer lacking the backbone ester groups. Both polymers presented similar molecular weights (A n values of 44.6 kDa vs 42.6 kDa, respectively) and dispersities (£> values of 1.38 vs 1.40, respectively). These values are similar to the theoretical molecular weights targeted for these reactionsAttorney Docket No. 370602-7086W01(00285)

[0408] (52.8 and 49.6 kDa, respectively). Collectively this indicates that introduction of esters via rROCCP did not compromise reaction control of the PET-RAFT chemistry.

[0409] The polymer pair was then subjected to chemical and enzymatic challenge to assess hydrolyzability of ester groups, with changes in molecular weight analyzed by size exclusion chromatography (SEC, FIG. 2A). Upon incubation with 50 mM NFUOH for 30 mins, a significant shift in the SEC profile was observed (Mn= 10.2 kDa, indicating approximately 4 degradable residues per chain), as well as a corresponding increase in dispersity (£> = 1.73) (FIG. 2B, red traces). This indicated the ester bonds in the backbone were rapidly hydrolyzed, leading to fragmentation of the polymer chain. In contrast, no change in molecular weight was observed for the DMA homopolymer lacking the ester residues in its backbone (FIG. 2B, black traces).

[0410] While alkaline-based degradation allows for straightforward assessment of the ability of the ester linkages to serve as sites of fragmentation in the backbone, intracellular lysosomal conditions are generally acidic, enzyme-rich environments. As such, based on these promising results, biodegradability’ of the polymers was then assessed using a model esterase (porcine liver esterase) to better represent lysosomal conditions. Importantly, the ester-containing copolymer demonstrated hydrolytic stability in the absence of enzyme, with minimal shift in the SEC trace over 48 h (FIG. 2C). However, when incubated with esterase, a gradual degradation of the polymer chain was observed (FIG. 2D, red traces). As expected, esterase had no effect on the molecular weight of the homopolymer (black traces), demonstrating that the ester groups incorporated by rROCCP could serve as sites of enzymatic degradation. Such stimuli-sensitive degradability' has clear utility’ for gene delivery’ vectors, as it allows robust extracellular stability while facilitating rapid disassembly and release of the nucleic acid payload upon endocytosis and trafficking to the esterase-rich lysosomal environment.

[0411] Example 2: Design and Synthesis of Exemplary Biodegradable Cationic Polymers Next, it was investigated whether this chemistry could be applied to polymer designs suitable for use as synthetic gene delivery vehicles. Specifically, a design based on an AEMAm-HEMA copolymer was selected as a model system due to previous reports of its success in this role A PET-RAFT polymerization was conducted with Cycl (10 mol% feed ratio), AEMAm (60 mol% feed ratio), and HEMA (30 mol% feed ratio) monomers to synthesize a biodegradable cationic polymer with a target degree of polymerization (DP) of 100 (FIG. 3). Time points were collected throughout the reaction and 'H NMR was used toAttorney Docket No. 370602-7086W01(00285)

[0412] independently monitor reaction kinetics of the three monomers over the course of the polymerization (FIG. 4A). The Cycl monomer was observed to convert less efficiently than the other monomers (only reaching 61.5% conversion after 18 h versus >99% and 89.7% for HEMA and AEMAm, respectively. However, when comparing Cycl conversion versus total monomer conversion (FIG. 4B), the macrocyclic monomer continues to undergo conversion throughout the course of the reaction, indicating ester groups are being incorporated throughout the length of the growing polymer chain. This distribution should facilitate significant fragmentation of the backbone upon degradation and could, in turn, improve payload release.

[0413] Next, a series of five PET-RAFT polymerizations of cationic copolymers were conducted, all targeting a theoretical DP of 100. but varying Cycl feed ratios in order to assess the relationship between monomer feed ratio and content of the degradable sequence in the copolymer product. Cationic comonomer content was held constant in the monomer feed (60 mol%), with Cycl and HEMA being varied from 0-10 mol% and 30-40 mol% respectively.1H NMR analysis of the copolymer products (FIGs. 9-13) indicated a clear linear relationship between feed ratio of the Cycl monomer and frequency of the degradable sequence in the backbone (FIG. 4C), suggesting that the degradability can be conveniently tuned simply by altering the feed ratio of the Cycl monomer. As observed with the polyDMA model system, incorporation of these ester groups allowed the polymer to undergo degradation, leading to fragmentation of the polymer backbone, with a higher density of degradable sequences yielding smaller polymer fragments (FIG. 22).

[0414] Example 3: Polyplex Formation and Transfection with Exemplary Backbone-Degradable Cationic Copolymers

[0415] A library of cationic AEMAm-HEMA copolymers (FIGs. 14-18 and Table 1) containing variable numbers of biodegradable residues were then evaluated for their ability to complex the model GFP plasmid (pMAX_GFP) and successfully transfect U-2 OS cells. Transfection efficiency was quantified using high-throughput cell imaging using a green fluorescence channel (483 / 536 nm) at a range of N / P ratios of 5, 10, and 20. N / P ratio is the stoichiometric ratio between the protonable nitrogen (N) in the copolymer and the anionic phosphate groups (P) present in the pMAX_GFP. Copolymers of AEMAm and HEMA at a DP of 100 have previously been shown to successfully delivery GFP gene payloads and induce significant expression. Furthermore, as the reported pKa of AEMAm-HEMA copolymers with similar charge density to those prepared herein is 8.2, this should result inAttorney Docket No. 370602-7086W01(00285)

[0416] significant protonation of the macromolecule at near-physiological pH. Therefore, the feed ratio of Cycl within this copolymer scaffold design was varied (60% AEMAm, 0-10% Cycl, 30-40 % HEM A) to establish that moderate backbone-degradability could be introduced without significant loss of function.

[0417] Table 1. Summary of monomer conversion of PET-RAFT reactions used to prepare polymer library (P1-P5) for in vitro transfection and cytotoxicity studies. _

[0418] Feed Ratio (mol%) % Conversion Poly ID [M]:[CTA]

[0419] HEMA AEMAm Cycl HEMA AEMAM Cycl Pl 100 30 60 10 99.7% 89.8% 48.0% P2 100 32.5 60 7.5 99.8% 90.9% 47.0% P3 100 35 60 5 99.7% 91.1% 48.1% P4 100 37.5 60 2.5 99.7% 90.5% 47.3%

[0420]

[0421] P5 100 40 60 - 99.6% 88.3% - Monomer conversion was determined based on assigned integrals for vinylic or allylic protons for each of the three unique monomers. Integrals were normalized to mesitylene internal standard (5 = 6.75 ppm), and change in the normalized integral (100% - I final / I initial) was used to calculate monomer conversion. Percent conversion reported for a monomer represents the average change across all integrals assigned to protons for that monomer.

[0422] Characterization of this polyplex library demonstrated these cationic polymers were able to successfully complex pDNA with high efficiency (FIG. 21), yielding well-defined polyplexes with relatively narrow dispersities and a lack of large aggregates (FIG. 20). FIG.

[0423] 5A shows the transfection efficiency obtained with the polyplexes formed with degradable copolymers. The transfection efficiency was calculated as the number of GFP-expressing cells divided by the total number of cells (obtained by Hoechst staining). It can be observed from FIG. 5Athat complexes formed with high Cycl feed ratios of 7.5% and 10% demonstrate the highest transfection efficiency at low N / P ratios (i.e.. low polymer content) of 5 and 10. However, at a higher N / P ratio of 20 (i.e.. at the highest copolymer concentration) their transfection efficiency significantly drops, owing to increased cytotoxicity. At lower Cycl content of 2.5 and 5 mol%, substantial transfection can still be observed at a high N / P ratio of 20. Complexes formed without Cycl (i.e.. non-backbone-degradable polymers) demonstrated a linear trend in transfection efficiency with increasing N / P ratio; however, their transfection was significantly lower than complexes formed with Cycl at low N / P ratios of 5 and 10 as observed in FIG. 5A. Specifically at N / P ratio of 5, a 10-fold increase in transfection is observed with complexes containing 7.5% and 10% Cycl compared to their non-backbone degradable analog.Attorney Docket No. 370602-7086W01(00285)

[0424] In sharp contrast, untreated cells and those treated with only the pMAX-GFP vector (i.e., no polymer) did not demonstrate any transfection (FIG. 19). This suggests that introducing backbone degradability could enhance the disassembly of the polyplex in the cytosol enabling more efficient payload release. Polyplexes were incubated with the cells for 4 h with serum-free media prior to introducing serum-containing media. Without wishing to be bound by any theory, this may occur because serum proteins can interfere with polyplex formation, causing early release of the plasmid and reducing its cellular uptake which may eventually lead to discrepancies between in vitro and in vivo results. FIG. 5B demonstrates the actual count of GFP-positive cells observed in each well.

[0425] Complexes formed from copolymers with high Cycl content (feed ratios of 7.5% and 10%) at a low N / P ratio of 5 again express more than twice the number of GFP-positive cells than that observed for polyplexes with lower Cycl content. However, at a slightly higher N / P ratio of 10, a lower Cycl content of 5% demonstrates higher GFP-positive cells than Cycl content of 7.5% and 10% which may be attributed negligible cell death as discussed in the next section. Further increasing the N / P ratio to 20 resulted in reduced GFP expression across all polyplexes as increasing Cycl content was not able to rescue expression at high polymer concentrations that can cause higher cellular toxicity. To investigate the amount of GFP molecules expressed in a single cell, the mean GFP intensity obtained from the Celigo image analysis software was plotted for the polyplex treated cells. No significant differences were observed in the intensity between the transfecting agents as observed in FIG. 5C except for Cycl content of 5% at N / P of 10 that demonstrated slightly higher intensity than the non-degradable copolymers at the same N / P. Therefore, it can be concluded that the ability of the polyplexes formed with degradable copolymers to produce GFP molecules in a cell is comparable to that of the commercial transfecting agent PEIpro.

[0426] Example 4: Cytotoxicity Evaluation of an Exemplary Polyplex Library

[0427] Next, the cause of cell death was assessed by the complexes formed with degradable copolymers. Although evaluating transfection efficiency is an important metric for deciding the best-performing transfecting agent, the fact that it is measured based on the total number of live cells is often overlooked. That is, if a transfecting agent is cytotoxic but can transfect as many cells as a non-cytotoxic transfecting agent, the cytotoxic agent will demonstrate higher efficiency due to the reduction in the total number of live cells. Therefore, assessing the cell death caused by transfecting agents is an important metric in deciding the bestperforming transfecting agent.Attorney Docket No. 370602-7086W01(00285)

[0428] The cell death caused by the degradable copolymers was assessed by counting the number of live cells at 48 h after transfection and compared it to the control cells. FIG. 6A demonstrates the cell death (%) which indicates that polyplexes formulated at a low N / P ratio of 5 demonstrate less than 20% cell death. The complex formed with 5% Cycl content at N / P ratio 5 did not demonstrate any cell death, that is, this formulation led to greater cell proliferation than the control cells. Complexes formed with 7.5% and 10% Cycl content at an N / P ratio of 10 had demonstrated high transfection efficiency (FIG. 5 A); however, it caused nearly 70% cell death, and therefore, its performance comes at the cost of unacceptable cytotoxicity. All the complexes formed at a high N / P ratio of 20, regardless of whether they are degradable or non-degradable, cause nearly 70-80% death compared to the control cells suggesting higher polymer content is toxic to the cells. Even PEIpro with high transfection efficiency causes a significant reduction in cell count or cell death leading to an artificial increase in the transfection efficiency. Along with degradability, Cycl also introduces hydrophobicity to the degradable copolymers due to its structure containing an eight-carbon aliphatic saturated chain.

[0429] The effects of hydrophobic modification on cationic polymers are controversial and studies have reported negative effects on the cytotoxicity of the gene delivery vector. In cationic RAFT copolymers, introducing hydrophobicity in the side chain is reported to elevate cytotoxicity. The increased cytotoxicity for degradable polyplexes with 7.5% and 10% Cycl content at N / P ratio of 10 may be attributed to the additional hydrophobicity Cycl introduced into the backbone. However, such effect is shown to be mitigated at the lower N / P ratios which also yield the highest transfection efficiency. Therefore, the effect of Cycl incorporation on the cytotoxicity of the polyplexes may be a more complex relationship than attributed to its hydrophobicity.

[0430] The viability of the cells was also assessed using a commercial CCK-8 kit. All polyplexes formed at a lower N / P ratio of 5 demonstrated nearly 100% cell viability which is higher than that demonstrated by PEIpro (75%) as shown in FIG. 6B. However, at higher N / P ratios of 10 and 20, the cell viability drops below 50% for most complexes formed with degradable copolymers. This demonstrates that polyplexes formed with higher Cycl content of 7.5% and 10% and a lower N / P ratio of 5 demonstrate higher transfection efficiency than their non-degradable analog while maintaining low cell death and high cell viability (FIG. 7). This suggests that these degradable copolymers have the potential to significantly improve biocompatibility while enhancing gene delivery function.Attorney Docket No. 370602-7086W01(00285)

[0431] Enumerated Embodiments

[0432] The following exemplary’ embodiments are provided, the numbering of which is not to be construed as designating levels of importance:

[0433] Embodiment 1 provides a polymer composition comprising a random copolymer of: (a) at least one hydrophilic vinyl monomer,

[0434] wherein the at least one hydrophilic vinyl monomer comprises a vinyl monomer substituted with at least one hydrophilic moiety;

[0435] (b) at least one cationic vinyl monomer,

[0436] wherein the at least one cationic vinyl monomer comprises a vinyl monomer substituted with at least one cationic moiety; and

[0437] (c) at least one cyclic monomer of formula (la):

[0438]

[0439] wherein:

[0440] Rlaand Rlbare each independently selected from the group consisting of H, optionally substituted Ci-Ce alkyl, and optionally substituted Ce-Cio aryl;

[0441] R2aand R2bare each independently selected from the group consisting of H, optionally substituted Ci-Ce alkyl, and optionally substituted Ce-Cio aryl;

[0442] R3aand R3bare each independently H;

[0443] L1comprises at least one divalent moiety’ selected from the group consisting of optionally substituted C1-C12 alkylenyl, optionally substituted C1-C12 heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted Ce-Cio arylenyl, and optionally substituted C2-C10 heteroarylenyl;

[0444] X1is selected from the group consisting of -O-*, -S-*. -S-S-*, -C(=O)O-*, -OC(=O)-*, -NHC(=O)-*, -C(=O)NH-*, -C(=S)O-*, -OC(=S)-*, -OC(=O)O-*, -NHC(=S)S-*, -SC(=S)NH-*, -NHC(=O)O-*, -OC(=O)NH-*, and a bond (absent);

[0445] X2is selected from the group consisting of **-O-, **-S-, **-S-S-, **-C(=O)O-, **-OC(=O)-, **-NHC(=O)-, **-C(=O)NH-, **-C(=S)O-, **-OC(=S)-, **-OC(=O)O-, **■ NHC(=S)S-, **-SC(=S)NH-, **-NHC(=O)O-, **-OC(=O)NH-, and a bond;

[0446] wherein no more than one of X1and X2is a bond;

[0447] Z1is selected from the group consisting of -S- and -S(=O)2-;Attorney Docket No. 370602-7086W01(00285)

[0448] * indicates a bond between X1and L1; and

[0449] ** indicates a bond between X2and L1.

[0450] Embodiment 2 provides the polymer composition of Embodiment 1, wherein one of the following applies:

[0451] (a) Rlais H and Rlbis CH3; or

[0452] (b) Rlais CHs and Rlbis H.

[0453] Embodiment 3 provides the polymer composition of Embodiment 1 or 2. wherein R2aand R2bare each independently selected from the group consisting of H and optionally substitute phenyl,

[0454] optionally wherein one of R2aand R2bis H, and one of R2aand R2bis Ph.

[0455] Embodiment 4 provides the polymer composition of any one of Embodiments 1-3, wherein X1is selected from the group consisting of -C(=O)O-* and -OC(=O)-*,

[0456] optionally wherein X1is -C(=O)O-*.

[0457] Embodiment 5 provides the polymer composition of any one of Embodiments 1-4, wherein X2is selected from the group consisting of **-OC(=O)- and **-C(=O)O-,

[0458] optionally wherein X2is **-OC(=O)-.

[0459] Embodiment 6 provides the polymer composition of any one of Embodiments 1-5, wherein L1is selected from the group consisting of -(CH2)-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)7-, -(CH2)8-, -(CH2)9-, -(CH2)10-, -(CH2)11-, and -(CH2)12-, optionally wherein L1is -(CH2)8-.

[0460] Embodiment 7 provides the polymer composition of any one of Embodiments 1-6, wherein Z1is -S-.

[0461] Embodiment 8 provides the polymer composition of any one of Embodiments 1-7, wherein the cyclic monomer of formula (la) is:

[0462]

[0463] Embodiment 9 provides the polymer composition of any one of Embodiments 1-8, wherein each hydrophilic monomer is independently a compound of formula (lb):

[0464] R4\ L2-R6

[0465] \= /

[0466] R

[0467]

[0468] 4bR5(lb),Attorney Docket No. 370602-7086W01(00285)

[0469] wherein:

[0470] R4aand R4bare each independently H;

[0471] R5is selected from the group consisting of H and Ci-Ce alkyl;

[0472] R6is selected from the group consisting of C1-C24 alkyl, C1-C24 heteroalkyl, Cs-Cs cycloalkyl, C2-C8 heterocycloalkyl, C6-C10aryl, and C2-C10 heteroaryl,

[0473] wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is substituted with at least one hydrophilic moiety, and

[0474] wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is further optionally substituted with at least one substituent selected from the group consisting of halogen, optionally substituted C i-Ce alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C10 heteroaryl;

[0475] L2comprises at least one divalent moiety selected from the group consisting of -O-, -N(RA)-, -C(=O)-. -C(=NRA)-, C(=S)-, -S(=O)-, and -S(=O)2-;

[0476] each occurrence of RAis independently selected from the group consisting of H, C(=O)RB, optionally substituted Ci-Ce alkyl, optionally substituted Ci-Ce heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroaryl; and each occurrence of RDis independently selected from the group consisting of optionally substituted Ci-Ce alkyl, optionally substituted Ci-Ce heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C10 heteroaryl.

[0477] Embodiment 10 provides the polymer composition of Embodiment 9. wherein R5is CH3.

[0478] Embodiment 11 provides the polymer composition of Embodiment 9 or 10, wherein L2is -C(=O)O-.

[0479] Embodiment 12 provides the polymer composition of any one of Embodiments 9-11. wherein each hydrophilic moiety is independently selected from the group consisting of ORA, C(=O)ORB, OC(=O)RA, OC(=O)ORB, OC(=O)SRA, OC(=O)N(RA)(RA), SRA, SC(=O)RA, SC(=O)ORB, SC(=O)N(RA)(RA), S(=O)RA, S(=O)ORB, S(=O)N(RA)(RA), S(=O)2RA, S(=O)2ORB, S(=O)2N(RA)(RA), N(RA)C(=O)RA, N(RA)C(=O)ORB, N(RA)C(=O)SRA, N(RA)C(=O)N(RA)(RA), N(RA)C(=S)RA, N(RA)C(=S)ORB, N(RA)C(=S)SRA.

[0480] N(RA)C(=S)N(RA)(RA), N(RA)S(=O)2RA, N(RA)S(=O)2RA, CN, NO2, halogen, and optionallyAttorney Docket No. 370602-7086W01(00285)

[0481] substituted C2-C10 heteroaryl,

[0482] optionally wherein at least one hydrophilic moiety is OH.

[0483] Embodiment 13 provides the polymer composition of any one of Embodiments 9-12, wherein R6is C1-C24 alkyl substituted with at least one OH,

[0484] optionally wherein R6is -(CH2)2OH.

[0485] Embodiment 14 provides the polymer composition of any one of Embodiments 1-13. wherein the hydrophilic vinyl monomer is

[0486] O

[0487]

[0488] Embodiment 15 provides the polymer composition of any one of Embodiments 1-14, wherein each cationic vinyl monomer is independently a compound of formula (Ic):

[0489] R7a |_3_R9

[0490]

[0491] R7bR8(Ic),

[0492] wherein:

[0493] R7aand R7bare each independently H;

[0494] R8is selected from the group consisting of H and Ci-Ce alkyl;

[0495] R9is selected from the group consisting of C1-C24 alkyl, C1-C24 heteroalkyl, Cs-Cs cycloalkyl, C2-C8 heterocycloalkyl, C6-C10aryl, and C2-C10 heteroaryl,

[0496] wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is substituted with at least one cationic moiety or ionizable (cationic) moiety, and

[0497] wherein the alkyd, heteroalkyl, cycloalkyl, heterocycloalkyd, aryl, or heteroaryl is further optionally substituted with at least one substituent selected from the group consisting of halogen, optionally substituted C1-C6 alkyl, optionally substituted Ci-Ce heteroalky7!, optionally substituted Cs-Cs cycloalkyd, optionally substituted C2-C8 heterocycloalkyl, optionally substituted Cd-Cio aryl, and optionally substituted C2-C10 heteroaryl;

[0498] L3comprises at least one divalent moiety selected from the group consisting of -O-, -N(RC)-, -C(=O)-, -C(=NRC)-, C(=S)-, -S(=O)-, and -S(=O)2-;

[0499] each occurrence of RCis independently selected from the group consisting of H, C(=O)RB, optionally substituted Ci-Ce alkyl, optionally substituted Ci-Ce heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C10 heteroaryl; andAttorney Docket No. 370602-7086W01(00285)

[0500] each occurrence of RDis independently selected from the group consisting of optionally substituted Ci-Ce alkyl, optionally substituted Ci-Ce heteroalkyl, optionally substituted C'3-Cs cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroaryl.

[0501] Embodiment 16 provides the polymer composition of Embodiment 15, wherein R8is CH3.

[0502] Embodiment 17 provides the polymer composition of Embodiment 15 or 16, wherein L3is -C(=O)NH-.

[0503] Embodiment 18 provides the polymer composition of any one of Embodiments 15-17, wherein at least one of the following applies:

[0504] (a) each cationic moiety is independently selected from the group consisting of - [NH?]+, -[NH2(RD)]+, -[NH(RD)2]+, -[N(RD)3]+, -[C(=NH2)NH2]+, - [C(=NHRD)NH2]+, -[C(=NH2)NHRD]+, -[C(=NH2)N(RD)2]+, - [

[0505]

[0506] C(=NHRD)NHRD]+, -[C(=NHRD)N(RD)2]+, guanidinium, pyridinium, pyrimidinium, pyrazinium, pyrrolium, imidazolium, triazolium, thiazolium. oxazolium, piperidinium, morpholinium. and pyrrolidinium. optionally wherein at least one cationic moiety is NH3+; and

[0507] (b) each ionizable (cationic) moiety is independently selected from the group consisting of NH2, NH(RD), N(RD)2, C(=NH)NH2, C(=NH)NHRD, - [C(=NH)N(RD)2, C(=NRD)NHRD, C(=NRD)N(RD)2, guanidinyl. pyridinyl, pyrimidinyl, pyrazinyl, pyrrolyl, imidazolyl, triazolyl, thiazolyl, piperidinyl, morpholinyl, and pyrrolidinyl,

[0508] optionally wherein the at least one ionizable (cationic) moiety is NH2.

[0509] Embodiment 19 provides the polymer composition of any one of Embodiments 1-17. wherein R9is C1-C24 alkyl substituted with at least one NH2 or NH3+, optionally wherein R9is -(CH2)2NH2or -(CH2)2NH3+.

[0510] Embodiment 20 provides the polymer composition of any one of Embodiments 1-19, wherein the cationic vinyl monomer is

[0511]

[0512] Embodiment 21 provides the polymer composition of any one of Embodiments 1-20. wherein the hydrophilic vinyl monomer comprises about 20 mol% to about 50 mol% of the polymer composition, optionally wherein the hydrophilic vinyl monomer comprises about 30Attorney Docket No. 370602-7086W01(00285)

[0513] mol% to about 40 mol% of the polymer composition.

[0514] Embodiment 22 provides the polymer composition of any one of Embodiments 1-21. wherein the cationic vinyl monomer comprises about 40 mol% to about 80 mol% of the polymer composition, optionally wherein the cationic vinyl monomer comprises about 60 mol% of the polymer composition.

[0515] Embodiment 23 provides the polymer composition of any one of Embodiments 1-22. wherein the cyclic monomer comprises about 0.1 mol% to about 20 mol% of the polymer composition, optionally wherein the cyclic monomer comprises about 1 mol% to about 10 mol% of the polymer composition.

[0516] Embodiment 24 provides the polymer composition of any one of Embodiments 1-23, wherein the hydrophilic vinyl monomer, cationic vinyl monomer, and cyclic monomer have a molar ratio selected from the group consisting of about 30:60:10, 32.5:60:7.5, 35:60:5, 37.5:60:2.5, 28.5:54.6:5, 30.9:54.6:3.6, 33.3:54.6:2.5, and 35.6:54.6:1.1.

[0517] Embodiment 25 provides a vector comprising the polymer composition of any one of Embodiments 1-24 and at least one nucleic acid cargo, wherein the polymer and nucleic acid cargo are non-covalently complexed.

[0518] Embodiment 26 provides the vector of Embodiment 25, w herein the nucleic acid cargo comprises RNA or DNA.

[0519] Embodiment 27 provides the vector of Embodiment 26, wherein the RNA is selected from the group consisting of messenger RNA (mRNA). small interfering RNA (siRNA). microRNA (miRNA), and small-guide RNA (sgRNA).

[0520] Embodiment 28 provides the vector of Embodiment 26, wherein the DNA is selected from the group consisting of plasmid DNA, genomic DNA, and synthetic DNA.

[0521] Embodiment 29 provides the vector of any one of Embodiments 25-28, wherein the vector has a N / P ratio ranging from about 1 / 1 to about 30 / 1, wherein the N / P ratio indicates a ratio of cationic charge (e.g., cationic nitrogen atoms) in the polymer composition (N) and anionic charge (e.g., anionic phosphate groups) in the nucleic acid cargo (P).

[0522] Embodiment 30 provides the vector of Embodiment 29, wherein the vector has a N / P ratio selected from the group consisting of about 5 / 1, about 10 / 1. and about 20 / 1.

[0523] Embodiment 31 provides a pharmaceutical composition comprising the vector of any one of Embodiments 25-30 and at least one pharmaceutically acceptable excipient.

[0524] Embodiment 32 provides a method of delivering a nucleic acid to a target cell, the method comprising administering to a subject comprising the target cell the vector of any one of Embodiments 25-30 and / or the pharmaceutical composition of Embodiment 31.Attorney Docket No. 370602-7086W01(00285)

[0525] Embodiment 33 provides a method for performing gene therapy in a subject, the method comprising administering to the subject the vector of any one of Embodiments 25-30 and / or the pharmaceutical composition of Embodiment 31.

[0526] Embodiment 34 provides a method for treating, ameliorating, and / or preventing a disease in a subject, the method comprising administering to the subject the vector of any one of Embodiments 25-30 and / or the pharmaceutical composition of Embodiment 31.

[0527] Embodiment 35 provides a method for modulating gene expression in a target cell, the method comprising administering to a subject comprising the target cell the vector of any one of Embodiments 25-30 and / or the pharmaceutical composition of Embodiment 31.

[0528] The terms and expressions employed herein are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the embodiments of the present application. Thus, it should be understood that although the present application describes specific embodiments and optional features, modification and variation of the compositions, methods, and concepts herein disclosed may be resorted to by those of ordinary skill in the art, and that such modifications and variations are considered to be within the scope of embodiments of the present application.

Claims

1. Attorney Docket No. 370602-7086W01(00285)CLAIMSWhat is claimed is:

1. A polymer composition comprising a random copolymer of:(a) at least one hydrophilic vinyl monomer,wherein the at least one hydrophilic vinyl monomer comprises a vinyl monomer substituted with at least one hydrophilic moiety;(b) at least one cationic vinyl monomer,wherein the at least one cationic vinyl monomer comprises a vinyl monomer substituted with at least one cationic moiety; and(c) at least one cyclic monomer of formula (la):wherein:Rlaand Rlbare each independently selected from the group consisting of H, optionally substituted Ci-Ce alkyl, and optionally substituted C6-C10 aryl;R2aand R2bare each independently selected from the group consisting of H, optionally substituted Ci-Ce alkyl, and optionally substituted Cs-Cio aryl;R3aand R3bare each independently H;L1comprises at least one divalent moiety selected from the group consisting of optionally substituted C1-C12 alkylenyl, optionally substituted C1-C12 heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted Ce-Cio arylenyl, and optionally substituted C2-C10 heteroarylenyl;X1is selected from the group consisting of -O-*, -S-*. -S-S-*. -C(=O)O-*, -OC(=O)-*, -NHC(=O)-*, -C(=O)NH-*, -C(=S)O-*, -OC(=S)-*, -OC(=O)O-*. -NHC(=S)S-*, -SC(=S)NH-*, -NHC(=O)O-*, -OC(=O)NH-*, and a bond (absent);X2is selected from the group consisting of **-O-, **-S-, **-S-S-, **-C(=O)O-, **-OC(=O)-, **-NHC(=O)-, **-C(=O)NH-, **-C(=S)O-, **-OC(=S)-, **-OC(=O)O-, **-NHC(=S)S-, **-SC(=S)NH-, **-NHC(=O)O-, **-OC(=O)NH-, and a bond;wherein no more than one of X1and X2is a bond;Z1is selected from the group consisting of -S- and -S(=O)2-;* indicates a bond between X1and L1; andAttorney Docket No. 370602-7086W01(00285)** indicates a bond between X2and L1.

2. The polymer composition of claim 1, wherein one of the following applies:(a) Rlais H and Rlbis CHs; or(b) Rlais CH3 and Rlbis H.

3. The polymer composition of claim 1 or 2, wherein R2aand R2bare each independently selected from the group consisting of H and optionally substitute phenyl,optionally wherein one of R2aand R2bis H, and one of R2aand R2bis Ph.

4. The polymer composition of any one of claims 1-3, wherein X1is selected from the group consisting of -C(=O)O-* and -OC(=O)-*,optionally wherein X1is -C(=O)O-*.

5. The polymer composition of any one of claims 1-4, wherein X2is selected from the group consisting of **-OC(=O)- and **-C(=O)O-,optionally wherein X2is **-OC(=O)-.

6. The polymer composition of any one of claims 1-5, wherein L1is selected from the group consisting of -(CH2)-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)7-, -(CH2)8-, -(CH2)9-, -(CH2)IO-, -(CH2)H-, and -(CH2)i2-,optionally wherein L1is -(CH2)s-.

7. The polymer composition of any one of claims 1-6, wherein Z1is -S-.

8. The polymer composition of any one of claims 1-7, wherein the cyclic monomer of formula (la) is:

9. The polymer composition of any one of claims 1-8, wherein each hydrophilicAttorney Docket No. 370602-7086W01(00285)monomer is independently a compound of formula (lb):R4aL2_R64b)=<R4bR5(lb),wherein:R4aand R4bare each independently H;R5is selected from the group consisting of H and Ci-Ce alkyl;R6is selected from the group consisting of C1-C24 alkyl. C1-C24 heteroalkyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, and C2-C10 heteroaryl,wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is substituted with at least one hydrophilic moiety, andwherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is further optionally substituted with at least one substituent selected from the group consisting of halogen, optionally substituted Ci-Ce alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroaryl;L2comprises at least one divalent moiety selected from the group consisting of -O-, -N(RA)-, -C(=O)-, -C(=NRA)-, C(=S)-, -S(=O)-, and -S(=O)2-;each occurrence of RAis independently selected from the group consisting of H, C(=O)RB, optionally substituted Ci-Ce alkyl, optionally substituted Ci-Ce heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C10 heteroaryl; and each occurrence of RBis independently selected from the group consisting of optionally substituted Ci-Ce alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C10 heteroaryl.

10. The polymer composition of claim 9, wherein R5is CH3.

11. The polymer composition of claim 9 or 10, wherein L2is -C(=O)O-.

12. The polymer composition of any one of claims 9-11, wherein each hydrophilic moiety is independently selected from the group consisting of ORA, C(=O)ORB, OC(=O)RA.Attorney Docket No. 370602-7086W01(00285)OC(=O)ORB, OC(=O)SRA, OC(=O)N(RA)(RA), SRA, SC(=O)RA, SC(=O)ORB, SC(=O)N(RA)(RA), S(=O)RA, S(=O)ORA. S(=O)N(RA)(RA), S(=O)2RA, S(=O)2ORB, S(=O)N(RA)(RA), N(RA)C(=O)RA, N(RA)C(=O)ORB, N(RA)C(=O)SRA, N(RA)C(=O)N(RA)(RA), N(RA)C(=S)RA, N(RA)C(=S)ORB, N(RA)C(=S)SRA, N(RA)C(=S)N(RA)(RA), N(RA)S(=O)2RA, N(RA)S(=O)2RA, CN, NO2, halogen, and optionally substituted C2-Cio heteroaryl,optionally wherein at least one hydrophilic moiety is OH.

13. The polymer composition of any one of claims 9-12, wherein R6is Ci-C24 alkyl substituted with at least one OH,optionally wherein R6is -(CH2)2OH.

14. The polymer composition of any one of claims 1-13, wherein the hydrophilic vinyl monomer is:O15. The polymer composition of any one of claims 1-14, wherein each cationic vinyl monomer is independently a compound of formula (Ic):R\ L3-R9R7bR8(ic),wherein:R7aand R7bare each independently H;R8is selected from the group consisting of H and Ci-Ce alkyl:R9is selected from the group consisting of Ci-C24 alkyl, Ci-C24 heteroalkyl, Cs-Cs cycloalkyl, C2-Cs heterocycloalkyl, C6-C10aryl, and C2-Cio heteroaryl,wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is substituted with at least one cationic moiety or ionizable (cationic) moiety, andwherein the alkyd, heteroalky l, cycloalky l, heterocycloalky l, ary 1, or heteroaryl is further optionally substituted with at least one substituent selected from the group consisting of halogen, optionally substituted Ci-Ce alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C?-Cs cycloalkyd,Attorney Docket No. 370602-7086W01(00285)optionally substituted C2-C8 heterocycloalkyl, optionally substituted Ce-Cio aryl, and optionally substituted C2-C10 heteroaryl;L3comprises at least one divalent moiety selected from the group consisting of -O-, -N(RC)-, -C(=O)-, -C(=NRC)-, C(=S)-, -S(=O)-, and -S(=O)2-;each occurrence of RC is independently selected from the group consisting of H, C(=O)RB, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C10 heteroaryl; and each occurrence of RD is independently selected from the group consisting of optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C10 heteroaryl.

16. The polymer composition of claim 15, wherein R8is CH3.

17. The polymer composition of claim 15 or 16, wherein L3is -C(=O)NH-.

18. The polymer composition of any one of claims 15-17, wherein at least one of the following applies:(a) each cationic moiety is independently selected from the group consisting of - [NH3]+, -[NH2(RD)]+, -[NH(RD)2]+, -[N(RD)3]+, -[C(=NH2)NH2]+, - [C(=NHRD)NH2]+, -[C(=NH2)NHRD]+, -[C(=NH2)N(RD)2]+, - [C(=NHRD)NHRD]+, -[C(=NHRD)N(RD)2]+, guanidinium, pyridinium, pyrimidinium, pyrazinium, pyrrolium, imidazolium, triazolium, thiazolium. oxazolium, piperidinium, morpholinium, and pyrrolidinium, optionally wherein at least one cationic moiety is NH3+; and(b) each ionizable (cationic) moiety is independently selected from the group consisting of NH2, NH(RD), N(RD)2, C(=NH)NH2, C(=NH)NHRD, - [C(=NH)N(RD)2, C(=NRD)NHRD, C(=NRD)N(RD)2. guanidinyl, pyridinyl, pyrimidinyl, pyrazinyl, pyrrolyl, imidazolyl, triazolyl, thiazolyl, piperidinyl, morpholinyl, and pyrrolidinyl,optionally wherein the at least one ionizable (cationic) moiety is NH2.

19. The polymer composition of any one of claims 1-17, wherein R9is Ci-C24 alkylAttorney Docket No. 370602-7086W01(00285)substituted with at least one NH2 or NH3+, optionally wherein R6 is -(CH2)2NH2 or -(CH2)2NH3+.

20. The polymer composition of any one of claims 1-19, wherein the cationic vinyl monomer is:O21. The polymer composition of any one of claims 1-20, wherein the hydrophilic vinyl monomer comprises about 20 mol% to about 50 mol% of the polymer composition, optionally wherein the hydrophilic vinyl monomer comprises about 30 mol% to about 40 mol% of the polymer composition.

22. The polymer composition of any one of claims 1-21, wherein the cationic vinyl monomer comprises about 40 mol% to about 80 mol% of the polymer composition, optionally wherein the cationic vinyl monomer comprises about 60 mol% of the polymer composition.

23. The polymer composition of any one of claims 1-22, wherein the cyclic monomer comprises about 0.1 mol% to about 20 mol% of the polymer composition, optionally wherein the cyclic monomer comprises about 1 mol% to about 10 mol% of the polymer composition.

24. The polymer composition of any one of claims 1-23, wherein the hydrophilic vinyl monomer, cationic vinyl monomer, and cyclic monomer have a molar ratio selected from the group consisting of about 30:60:10, 32.5:60:7.5, 35:60:5, 37.5:60:2.5, 28.5:54.6:5, 30.9:54.6:3.6, 33.3:54.6:2.5, and 35.6:54.6:1.1.

25. A vector comprising the polymer composition of any one of claims 1 -24 and at least one nucleic acid cargo, wherein the polymer and nucleic acid cargo are non-covalently complexed.

26. The vector of claim 25, wherein the nucleic acid cargo comprises RNA or DNA.Attorney Docket No. 370602-7086W01(00285)27. The vector of claim 26, wherein the RNA is selected from the group consisting of messenger RNA (mRNA). small interfering RNA (siRNA). microRNA (miRNA), and smallguide RNA (sgRNA).

28. The vector of claim 26, wherein the DNA is selected from the group consisting of plasmid DNA, genomic DNA, and synthetic DNA.

29. The vector of any one of claims 25-28, wherein the vector has a N / P ratio ranging from about 1 / 1 to about 30 / 1, wherein the N / P ratio indicates a ratio of cationic charge (e.g., cationic nitrogen atoms) in the polymer composition (N) and anionic charge (e.g., anionic phosphate groups) in the nucleic acid cargo (P).

30. The vector of claim 29, wherein the vector has a N / P ratio selected from the group consisting of about 5 / 1, about 10 / 1, and about 20 / 1.

31. A pharmaceutical composition comprising the vector of any one of claims 25-30 and at least one pharmaceutically acceptable excipient.

32. A method of delivering a nucleic acid to a target cell, the method comprising administering to a subject comprising the target cell the vector of any one of claims 25-30 and / or the pharmaceutical composition of claim 31.

33. A method for performing gene therapy in a subject, the method comprising administering to the subject the vector of any one of claims 25-30 and / or the pharmaceutical composition of claim 31.

34. A method for treating, ameliorating, and / or preventing a disease in a subject, the method comprising administering to the subject the vector of any one of claims 25-30 and / or the pharmaceutical composition of claim 31.

35. A method for modulating gene expression in a target cell, the method comprising administering to a subject comprising the target cell the vector of any one of claims 25-30 and / or the pharmaceutical composition of claim 31.

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