Polymer Delivery Systems

Abstract

The polymeric delivery system delivers a biologic to a cell. In some embodiments, the polymeric delivery system comprises a polyplex. Each polyplex comprises at least one charged polymer and at least one biologic. The at least one charged polymer comprises a polyester copolymer of a polyol and a polycarboxylic acid, modified with at least one charged moiety having a net charge opposite to that of the at least one biologic. In other embodiments, the polymeric delivery system comprises a self-assembled particle comprising a block copolymer and a biologic bound to the block copolymer. The block copolymer comprises a first block of a polyester copolymer of a polyol and a polycarboxylic acid and a second block of a second monomer or second polymer.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63 / 271,275, both filed on October 25, 2021, and U.S. Provisional Patent Application No. 63 / 271,283, both of which are incorporated by reference in their entireties herein.

[0002]

[0002] The present disclosure is directed generally to polymeric delivery systems. More specifically, the present disclosure is directed to polymeric delivery systems for delivering biologics to biological systems, the polymers comprising polyester copolymers of polyols and polycarboxylic acids. [Background technology]

[0003]

[0003] Efficient targeted delivery of nucleic acids is a goal in many biological fields. Cationic lipid compounds have been used to deliver nucleic acids to living cells, for example, by transfecting cells in vitro for research. Nucleic acids are highly negatively charged, making them difficult to cross the cell membrane and difficult to escape from the endosomal compartments that form when they are taken up into cells. Nucleic acids are also cleaved by nucleases that can be present both extracellularly and intracellularly.

[0004]

[0004] Polyplexes, more specifically cationic polymer / therapeutic gene complexes, show potential for efficient delivery of bioactive compounds to cells, including mammalian cells. For example, cationic polymers mixed with plasmid deoxyribonucleic acid (pDNA) form binary polyelectrolyte complexes that can be taken up by cells, but these formulations have limited use due to their short circulation time in vivo and cytotoxicity at high concentrations.

[0005]

[0005] Although certain cationic polymers are very efficient in condensing nucleic acids and protecting them from degradation by nucleases, cationic polymers may also be toxic to cells depending on the total amount of net positive charge present and the properties of the polymer, such as molecular weight or degree of branching. Cationic polyamines, such as polyethyleneimine (PEI), poly(L-lysine), polyamidoamines, chitosan, polyaminoesters, and polyacrylates, have been evaluated as promising nucleic acid delivery vehicles.

[0006] Over the past two decades, nanomedicine and nanoparticle-based targeted delivery of biologic payloads for therapeutic administration has had a history of mixed success. Reasons for lackluster success include a lack of understanding of the reticuloendothelial and immune systems that keep foreign bodies out of the human body at the submicron level, a limited understanding of monocyte and macrophage differentiation, and an inadequate understanding of material surface and cellular interactions. For example, only about 0.7% of an injected nanoparticle dose reaches the target tumor (see, e.g., Mitchell et al., “Engineering precision nanoparticles for drug delivery,” Nature Reviews: Drug Discovery, Vol. 20, pp. 101-124 (2021)). Over the past decade or so, medical engineers and biomedical scientists may have been unaware of the impact of the immune system on parenteral administration, whether subcutaneous, intravenous, intramuscular, or intrathecal, and this lack of awareness may have led to confusion and overgeneralization of success.

[0007]

[0007] Both routes of administration attack both the humoral and cellular components of the human immune system, so there is a need to better understand the impact of biomaterials on immune-mediated responses to achieve greater success in administration and transplantation therapy, including cell therapy, gene therapy, drug delivery, and transplantation techniques. The biological responses to traditional synthetic biodegradable polymers, such as lactides, glycolides, anhydrides, fumarates, and phosphazenes, are known and must be taken into account when planning formulations using these materials. Thus, due to the lack of availability of non-immunogenic materials, some degree of inflammatory response or lack of biocompatibility must be accepted when formulating small particle delivery for human therapeutic treatments using such materials. The lack of such non-immunogenic materials has made recent attempts to overcome the challenges of particle delivery simply difficult.

[0008]

[0008] Conventional non-viral gene delivery systems are typically either lipid nanoparticles or polyplexes. Liposomes, micelles, polymeric micelles, polymersomes, dendrimers, and niosomes are nanoparticle systems that can each be the basis of gene delivery systems. Liposomes contain amphiphilic lipid molecules, typically two-tailed phospholipids with charged hydrophilic heads, that self-assemble into spherical structures with a lipid bilayer shell surrounding an aqueous core containing a hydrophilic payload, e.g., nucleic acid. Micelles contain amphiphilic lipid molecules that self-assemble into a single-layered spherical vesicular structure with polar head groups surrounding a hydrophobic tail core. Polymeric micelles are based on block copolymers with cationic and uncharged hydrophobic groups, and self-assemble with nucleic acids to form a core-shell structure. Polymersomes, like liposomes, are self-assembled structures composed of amphiphilic block copolymers. Dendrimers are highly branched spherical polymers that may be highly charged. Niosomes are spherical vesicular structures similar to liposomes, except that they are typically formed from non-ionic surfactants that have a single hydrophobic tail.

[0009]

[0009] Lipid nanoparticles (LNPs) represent a class of particles formed by amphiphilic structures, including but not limited to liposomes and micelles, that encapsulate payloads in either hydrophilic or hydrophobic cores, layers, or shells. Lipid nanoparticles are formed by the self-assembly of amphiphilic lipid molecules into globular structures. Most lipids traditionally used to form LNPs have cationic polar head groups required for binding to negatively charged nucleic acids. The tail portion of the lipid is typically one or more alkyl chains, which can vary in length and saturation. Other functional groups can be introduced depending on the desired functional properties of the LNPs.

[0010]

[0010] Conventional lipid nanoparticles typically contain polyethylene glycol (PEG) in some polar head groups and are loaded with biologics in the aqueous core. Concerns with conventional LNPs in nanoparticle delivery systems include the instability of the nanoparticles under physiological conditions due to charge screening interactions. Furthermore, the presence of PEGylated lipids that mask LNPs from the innate immune system may cause allergic reactions and enhance immune responses upon repeated exposure.

[0011] Polyplexes are nanoparticles formed and bound by electrostatic interactions between charged polymers and oppositely charged target molecules. Cationic polymers traditionally used to form polyplexes contain multiple amine functional groups that can carry a positive charge at physiological pH. Cationic polymers are routinely used as transfection agents to form polyplexes with negatively charged nucleic acids to facilitate the transfer of nucleic acids across cell membranes. Once inside the cell, the polyplexes dissociate, releasing the nucleic acid to complete the delivery and perform their function. One concern with cationic poly(amines) such as PEI is that they are cytotoxic at high concentrations, limiting their delivery capabilities and in vivo use. Other polymer-based transfection agents are also non-absorbable, requiring the use of additional cellular energy to remove the polymer from cells that are likely already in poor health as a result of their interaction with polyplex components. Because cell therapy is based on the use of cells taken from diseased and immunocompromised patients, such additional stresses can reduce the yield and quality of the final cell product.

[0012]

[0012] Polymersomes are another class of polymeric carriers that are similar to liposomes in terms of self-assembly, but are formed from amphiphilic polymers rather than molecular lipids. Polymersomes offer additional advantages over liposomal structures in that the larger molecular weight of the polymers allows for the encapsulation of larger payloads and more flexible modification of charge-hydrophobic properties. In some cases, polymer structures also offer additional biocompatibility over small molecular lipids. Summary of the Invention [Problem to be solved by the invention]

[0013]

[0013] There is a need for more biocompatible gene delivery systems that offer the same or greater ability to deliver biological payloads than current nanoparticle delivery systems. [Means for solving the problem]

[0014] In an exemplary embodiment, the polymeric delivery system comprises nanoparticles, each nanoparticle comprising at least one polymer and at least one biologic, wherein the at least one polymer comprises a polyester copolymer of a polyol and a polycarboxylic acid.

[0015] In an exemplary embodiment, the polymeric delivery system comprises a polyplex. Each polyplex comprises at least one charged polymer and at least one biologic. The at least one charged polymer comprises a polyester copolymer of a polyol and a polycarboxylic acid modified with at least one charged moiety having a net charge opposite to that of the at least one biologic.

[0016] In an exemplary embodiment, the polymeric delivery system includes a self-assembled particle comprising a block copolymer and a biologic associated with the block copolymer, the block copolymer including a first block of a polyester copolymer of a polyol and a polycarboxylic acid and a second block of a second monomer or second polymer.

[0017] In an exemplary embodiment, a method for forming a polymeric delivery system includes combining a macromolecule and a biologic in a solvent to form nanoparticles. Each nanoparticle includes at least one polymer and at least one biologic. Each polymer includes a polyester copolymer of a polyol and a polycarboxylic acid.

[0018] In an exemplary embodiment, a method for forming a polymeric delivery system comprising a polyplex includes combining a charged polymer and a biologic in a solvent to electrostatically associate the charged polymer and the biologic as a polyplex. Each polyplex includes at least one charged polymer and at least one biologic. Each charged polymer includes a polyester copolymer of a polyol and a polycarboxylic acid modified with at least one charged moiety having a net charge opposite to that of the biologic.

[0019] In an exemplary embodiment, a method for forming a polymeric delivery system includes forming self-assembled particles in a solvent that include a block copolymer and a biologic bound to the block copolymer, the block copolymer including a first block of a polyester copolymer of a polyol and a polycarboxylic acid and a second block of a second monomer or second polymer.

[0020] In an exemplary embodiment, a method for delivering a biologic to a cell includes administering nanoparticles comprising a polymer and the biologic, The polymer comprises a polyester copolymer of a polyol and a polycarboxylic acid.

[0021] In an exemplary embodiment, a method for delivering a biologic to a cell includes administering a polyplex of the biologic electrostatically associated with a charged polymer, the charged polymer comprising a polyester copolymer of a polyol and a polycarboxylic acid modified with at least one charged moiety having an opposite charge to the net charge of the biologic.

[0022] In an exemplary embodiment, a method for delivering a biologic to a cell includes administering a self-assembled particle including a block copolymer and a biologic bound to the block copolymer, the block copolymer including a first block of a polyester copolymer of a polyol and a polycarboxylic acid and a second block of a second monomer or second polymer. [Brief description of the drawings]

[0023] [Figure 1] FIG. 2 is a schematic diagram illustrating the formation of a charged polymer from a polyester copolymer of a polyol and a polycarboxylic acid and a charged moiety in an embodiment of the present disclosure. [Diagram 2]

[0024] FIG. 1 is a schematic diagram showing the electrostatic formation of a charged polymer-nucleic acid polyplex delivery system in an embodiment of the present disclosure. [Diagram 3]

[0025] FIG. 1 is a schematic diagram of a polymeric micelle of a biologic and a block copolymer in an embodiment of the present disclosure. [Figure 4]

[0026] FIG. 1 is a schematic diagram of a polymersome of a biologic and a block copolymer in accordance with an embodiment of the present disclosure. [Figure 5A]

[0027] FIG. 1 shows the attached proton test (APT) 13C-NMR spectrum of poly((glycerol sebacate 10% succinate 90%)-co-xylitol). [Figure 5B]

[0028] FIG. 13 shows the APT 13C-NMR spectrum of poly((glycerol sebacate 10% succinate 90%)-co-xylitol dimethylaminopropylamine). [Figure 6]

[0029] FIG. 1 shows Fourier transform infrared attenuated total reflectance (FTIR-ATR) spectra of poly((glycerol sebacate 10% succinate 90%)-co-xylitol) and poly((glycerol sebacate 10% succinate 90%)-co-xylitol dimethylaminopropylamine). [Figure 7]

[0030] FIG. 13 shows images obtained by fluorescence microscopy demonstrating successful transfection of primary human cardiac fibroblasts after 48 hours of exposure to polyplexes, indicated by the bright areas of the images. [Figure 8]

[0031] FIG. 1 shows images obtained by fluorescence microscopy demonstrating successful transfection of mouse NIH / 3T3 cells after 48 hours of exposure to polyplexes, indicated by the bright areas of the images. [Figure 9]

[0032] FIG. 1 shows images obtained by fluorescence microscopy demonstrating successful transfection of mouse NIH / 3T3 cells after 48 hours of exposure to polyplexes, indicated by the bright areas of the images. [Figure 10]

[0033] FIG. 13 shows images obtained by fluorescence microscopy demonstrating successful transfection of Jurkat E6.1 cells after 24 hours of exposure to polyplexes, indicated by the bright areas of the images. [Figure 11]

[0034] 1 is a graph showing the particle size distribution of polyplex particles with a weight ratio of mRNA:charged polymer of 1:40 in an embodiment of the present disclosure. [Figure 12]

[0035] 1 is a graph showing the zeta potential of polyplex particles with a weight ratio of mRNA:charged polymer of 1:40 in an embodiment of the present disclosure. [Figure 13]

[0036] 1 is a graph showing the particle size distribution of polyplex particles with a weight ratio of pDNA:charged polymer of 1:50 in an embodiment of the present disclosure. [Figure 14]

[0037] 1 is a graph showing the zeta potential of polyplex particles with a weight ratio of pDNA:charged polymer of 1:50 in an embodiment of the present disclosure. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024]

[0038] Wherever possible, the same reference numbers will be used throughout the drawings to represent the same parts.

[0025]

[0039] A polymeric delivery system is provided that includes a polymer and nanoparticles of a biologic bound to the polymer, where the polymer includes a polyester copolymer of a polyol and a polycarboxylic acid.

[0026]

[0040] Embodiments of the present disclosure provide polymers that are non-immunogenic, for example, compared to concepts that do not include one or more of the features disclosed herein; provide charged polymer structures that can be designed to fit a range of payload sizes for targeted delivery, tailored to specific degradation rates, and tuned to create a range of structures that can be decorated with specific functional moieties; utilize the immune stealth properties of poly(glycerol sebacate) (PGS) to reduce or avoid the presence of PEGylated lipids in the delivery system; utilize the low inflammatory response to polymer degradation to provide absorbable gene delivery vehicles that can be easily metabolized in cells to reduce cell stress; improve nanoparticle stability to extend blood residence time; provide a polymer platform with customizable polymer structures that address limitations of conventional gene delivery systems; provide block copolymer structures that can be tailored to create a range of polymer structures that can be designed to fit a range of payload sizes for targeted delivery, tailored to specific degradation rates, and tuned to create a range of polymer structures that can be decorated with specific functional moieties; provide a delivery system for biologics with reduced inflammatory response; provide absorbable polymer delivery vehicles that can be easily metabolized in cells to reduce cell stress; improve nanoparticle stability to extend blood residence time; or combinations thereof.

[0027]

[0041] As used herein, "polyester copolymer of polyol and polycarboxylic acid" refers to any copolymer that includes alternating monomer units of one or more polyols and one or more polycarboxylic acids linked by ester bonds. Such copolymers may include additional monomers that may not be polyols or polycarboxylic acids, as well as additional polymer blocks that may or may not include polyol and / or polycarboxylic acid monomers. Such copolymers may also include other functional groups. In some embodiments, the polyester copolymer of polyol and polycarboxylic acid includes glycerol and sebacic acid monomers.

[0028]

[0042] As used herein, "polyol" refers to any monomer having more than one alcohol group.

[0029]

[0043] As used herein, "polycarboxylic acid" refers to any monomer having more than one carboxylic acid group.

[0030]

[0044] As used herein, a "biologic" refers to any compound or component that is based on biology and that is desirable to deliver to and interact with a biological system. Exemplary biologics include, but are not limited to, nucleic acids, proteins, peptides, a gene editing system, antibodies, cytokines, and active pharmaceutical ingredients (APIs). Biologics may be of natural or synthetic origin, including, but not limited to, synthetic or recombinant nucleic acids, synthetic peptides, or recombinant proteins. Nucleic acid biologics may be single-stranded or double-stranded, and may be deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). Suitable gene editing systems include, but are not limited to, clustered regularly interspaced short palindromic repeats (CRISPR) systems, such as CRISPR / Cas9, transposons, transcription activator-like effector nucleases (TALENs), or zinc finger nucleases (ZFNs).

[0031]

[0045] In an exemplary embodiment, the polymer of the polymeric delivery system has Formula (1), shown below: [[-A a -B b ] x -C c -D d ] z (1) where A is a polyol monomer, a=1-30, B is a polycarboxylic acid monomer, b=1-30, C is a polymer block, charged moiety, or polyol monomer, c=0-30, D is a polymer block, charged moiety, or polycarboxylic acid monomer, d=0-30, x is an integer from 1 to 75 or any integer value, range, or subrange therebetween, and z is an integer from 1 to 100 or any integer value, range, or subrange therebetween. Includes the structure of.

[0032]

[0046] When present, C and / or D can be customized and tailored to the particular chemical and / or physical properties of the copolymer. In some embodiments, A is conjugated to a biologic. In other embodiments, B is conjugated to a biologic. In other embodiments, C is conjugated to a biologic. In other embodiments, D is conjugated to a biologic. In some embodiments, where C and / or D in formula (1) are polymer blocks, the polymer block is a polyacid, polyol, or polyamine. Suitable polymers may include, but are not limited to, PEG, PEI, polylysine, polyvinyl alcohol, polyvinyl acetate, hyaluronic acid, or aggrecan.

[0033]

[0047] In an exemplary embodiment, the polymer is coupled to the biologic through ligand interactions or complex coordination.

[0034]

[0048] In some embodiments, the polyester copolymers of polyols and polycarboxylic acids contain low concentrations of urethanes and / or acrylates that do not affect the solubility of the polymer.

[0035]

[0049] In some embodiments, the polyester copolymer of polyol and polycarboxylic acid includes glycerol as the polyol and sebacic acid as the polycarboxylic acid. The glycerol ester portion may be considered a lipid-like bis-diacid triglyceride of the polymer. Furthermore, these glycerol esters are polymers constructed from metabolic building blocks that feed directly into the Krebs cycle upon degradation or erosion. Thus, an underappreciated advantage of polymers such as PGS is the favorable local cell-substance interactions and immune responses. In contrast, degradable polymers composed of lactide and glycolide initiate immune responses with inflammatory macrophages (M1) due to the release of lactic and glycolic acids as degradation by-products. Furthermore, PGS can induce stimulation of oxidative phosphorylation due to the presence of glycerol ester metabolic by-products that initiate pro-healing (M2) macrophage differentiation. The presence of M2 is essential to alleviate, suppress, and / or control local inflammation during therapeutic delivery, allowing therapy to be initiated without complications and disruptions from undesirable substances, acid by-products, initiating immune responses.

[0036]

[0050] In some embodiments, the polymer is derivatized with a ligand for targeted payload delivery, for example at the first block or alcohol of the polyol, or other tether moiety. The ligand is designed to bind to a specific cell marker, which may include, but is not limited to, a tumor-specific antigen, a tumor-associated antigen protein, an antibody, alpha-fetoprotein, carcinoembryonic antigen, cancer antigen 125 (CA-125), mucin 1 (MUC-1), epithelial tumor antigen, tyrosinase, melanoma-associated antigen, aberrant products of ras, tumor protein P53 (p53), programmed cell death protein 1 (PD-1), major histocompatibility complex (MHC) protein, or a cell-specific epitope.

[0037] Charged Polymer

[0051] In some embodiments, the polymer is a charged polymer. In such embodiments, the C unit, if present, is a polyol monomer, or the D unit, if present, is a polycarboxylic acid monomer. The charged polymer further comprises at least one charged moiety, which may be a C unit, a D unit, or a pendant group on an A unit, a B unit, a C unit, and / or a D unit.

[0038]

[0052] In some embodiments, the polyester copolymer of polyol and polycarboxylic acid is a condensation reaction product of one or more polyol monomers and one or more polycarboxylic acid monomers.

[0039]

[0053] Suitable polyol monomers may include, but are not limited to, glycerol, low molecular weight PEG (about 1000 Da or less), polyvinyl alcohol, xylitol, mannitol, sorbitol, maltitol, erythritol, or isomalt. When more than one polyol monomer is copolymerized with a polycarboxylic acid monomer, they may be included in any molar ratio ranging from 1:99 to 50:50.

[0040]

[0054] Suitable polycarboxylic acid monomers include, but are not limited to, those having the general formula [HOOC(CH2) m Z(CH2) nCOOH] [wherein m and n=1-30, Z is -O-, -COO-, -CO-, -SS-, -NH-, -NZ'-, -CHZ"-, -CH=CH-, -C≡C-, or -CH=CH-CH=CH-, Z' is -H, -CH3, -CH2CH3, -CH2CH2NH2, or -CH2CH2NHCH3, and Z" is -H, -OH, -CH2OH, -CH2CH2OH, -CH2CH2NH2, or -CH2CH2NHCH3]. Suitable polycarboxylic acid monomers may include malonic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, oxalacetic acid, citric acid, fumaric acid, or malic acid. When more than one polycarboxylic acid monomer is copolymerized with a polyol monomer, they may be present in any molar ratio ranging from 1:99 to 50:50.

[0041]

[0055] Other suitable monomers in the charged polymer may include, but are not limited to, glutamine, lysine, or arginine.

[0042]

[0056] In some embodiments, the polyester copolymer of polyol and polycarboxylic acid is non-linear and / or contains branches and forms a charged polymer when modified with a charged moiety. In some embodiments, the polyester copolymer of polyol and polycarboxylic acid has a low to medium molecular weight. In exemplary embodiments, the weight average molecular weight of the polyester copolymer of polyol and polycarboxylic acid is about 2 kDa to about 50 kDa, alternatively about 2 kDa to about 30 kDa, alternatively about 2 kDa to about 15 kDa, alternatively about 2 kDa to about 10 kDa, alternatively about 5 kDa to about 50 kDa, alternatively about 10 kDa to about 50 kDa, alternatively about 10 kDa to about 25 kDa, or any value, range, or subrange therebetween. In exemplary embodiments, the polyester copolymer of polyol and polycarboxylic acid is not in a thermoset state. In some embodiments, the polyester copolymer of polyol and polycarboxylic acid has a high polydispersity index.

[0043]

[0057] In an exemplary embodiment, a polyester copolymer of a polyol and a polycarboxylic acid is modified with at least one charged moiety to form a charged polymer.

[0044]

[0058] In an exemplary embodiment, the polyester copolymer of a polyol and a polycarboxylic acid contains two or more different polycarboxylic acids and is modified with at least one charged moiety to form a charged polymer.

[0045]

[0059] In an exemplary embodiment, the polyester copolymer of a polyol and a polycarboxylic acid contains two or more different polyols and is modified with at least one charged moiety to form a charged polymer.

[0046]

[0060] In an exemplary embodiment, the polyester copolymer of polyol and polycarboxylic acid contains two or more different polycarboxylic acid monomers, two or more different polyol monomers, and is modified with at least one charged moiety to form a charged polymer.

[0047]

[0061] 1, a polyester copolymer 10 of polyol and polycarboxylic acid includes multiple R functional groups. In some embodiments, the R groups include hydroxyl groups, carboxyl groups, amine groups, or combinations thereof, such that the R groups are tether sites for charged compounds 12. Charged compounds 12 are represented by R'-X + 1, where R′ group represents complimentary chemistry to the tether moiety R, and X +represents a charged moiety. In some embodiments, the complementary chemically reactive moieties of the R' group include carboxyl groups that react with hydroxy and / or amine groups, isocyanate groups that react with hydroxy and / or amine groups, hydroxy groups that react with carboxyl groups, amine groups that react with carboxyl groups, acid chlorides that react with hydroxy and / or amine groups, ring-opening reactions, or combinations thereof. In some embodiments, activators such as, for example, carbonyl diimdazole or tosyl compounds may be used. Through this reaction scheme, polyester copolymers 10 of polyols and polycarboxylic acids can be derivatized to produce charged polymers 14. Charged compounds 12 may be introduced via monomers that copolymerize with the polyol and polycarboxylic acid monomers and are located within the repeat units of the polymer, or charged compounds 12 may be pendant by introduction after copolymerization of the polyol and polycarboxylic acid monomers.

[0048]

[0062] Suitable charged compounds may include, but are not limited to, amino acids such as glutamine, lysine, or arginine, dimethylaminopropanoic acid, 1,1-dimethylethylenediamine, PEI, polylysine, or hyaluronic acid.

[0049]

[0063] Suitable charged moieties may include, but are not limited to, sulfate, phosphate, sulfonate, sulfite, carboxylate, carbohydrate, glycoprotein, ammonium, amino acid, peptide, peptide sequence, e.g., arginine-glycine-aspartic acid (RGD) sequence that promotes cell adhesion, choline, phosphocholine, primary amine, secondary amine, tertiary amine, quaternary amine, or carboxylate.

[0050] Block Copolymers

[0064] In other embodiments, the polymer is a block copolymer. In some embodiments, the AB group of the block copolymer of formula (1) is a group represented by formula (2): -[-CO-(CH2) m-R-(CH2) n -CO2-CH2-CH(OR')-CH2-O-] x (2) [Wherein, A is -CH2-CH(OR')-CH2-O- and B is -CO-(CH2) m -R-(CH2) n -CO-, m and n are integers from 1 to 30 or any integer value, range, or subrange therebetween, R is -CH2-, -NH-, -NCH3-, -O-, -SS-, -CH=CH-, -C≡C-, -CO2-, or -CONH-, and R' is hydrogen or another interchangeable pendant group. wherein the C and / or D groups of formula (1) are customizable monomer or polymer blocks that can be tailored for specific chemical and physical properties of the block copolymer.

[0051]

[0065] In some embodiments, the first block of the block copolymer of formula (1) is produced by reaction of a diepoxide with a polyol. In some embodiments, the polyol is a diol. Formula (3) is an example of a diepoxide and a polyol that can react to form a polyester copolymer: H2COCH-CH2-O2C-(CH2) n -CO2-CH2-CHOCH2+HO-CH2-(CHR) x -(CHR') y -CH2-OH (3) wherein n is an integer from 1 to 20, or any integer value, range, or subrange therebetween; x is an integer from 1 to 10, or any integer value, range, or subrange therebetween; y is an integer from 0 to 10, an integer from 1 to 10, or any integer value, range, or subrange therebetween; and R and R' may independently represent -H or -OH. Such diepoxide-diol monomer pairs can provide substantially unbranched polymers.

[0052]

[0066] In other embodiments, the second block of the block copolymer of formula (1) is produced by reaction of a diepoxide with a polyamine. In some embodiments, the polyamine is a diamine. Formula (4) is an example of a diepoxide and a polyamine that can react to form a polyamine copolymer: H2COCH-CH2-O2C-(CH2) n -CO2-CH2-CHOCH2+H2N-(CH2) x -X m -(CH2) y -NH2(4) where n is an integer from 1 to 20, or any integer value, range, or subrange therebetween; m is an integer from 0 to 10, or any integer value, range, or subrange therebetween; x and y are independently integers from 0 to 10, or any integer value, range, or subrange therebetween; and X is selected to provide the AB block with particular properties, such as a predetermined charge density, a predetermined hydrophobicity, or a predetermined hydrophilicity. Suitable X groups may include, but are not limited to, -CH-, -NH-, -CH-CH-O-, -CH-CH-NH-, -CH-CH-CH-CH-NH-, or phenyl. Such diepoxide-diamine monomer pairs can provide substantially unbranched polymers.

[0053]

[0067] In some embodiments, the second block of C and / or D in formula (1) is selected to render the block copolymer amphiphilic. In some embodiments, the second block is selected to render the copolymer hydrophilic. In some embodiments, the second block is selected to render the polymer hydrophobic. In some embodiments, the second block is selected to render the resulting copolymer a branched copolymer. In some embodiments, the second block is selected to crosslink the resulting copolymer. In some embodiments, the second block is selected to render the resulting copolymer cationic. In some embodiments, the second block is selected to render the resulting copolymer anionic.

[0054]

[0068] In some embodiments, the second block has a cationic charge, allowing electrostatic complexation with negatively charged biologics, such as nucleic acids, while in other embodiments, the second block has an anionic charge, allowing electrostatic complexation with positively charged biologics.

[0055]

[0069] In an exemplary embodiment, the second block is more hydrophilic than the polyester copolymer of polyol and polycarboxylic acid, and therefore can be dissolved in an aqueous solution. In some embodiments, the values ​​of a, b, c, d, and z in formula (1) are selected such that the block copolymer self-assembles in a particular aqueous environment. In some embodiments, the second block is polyglycerol. In some embodiments, the second block is poly(vinyl alcohol). In some embodiments, the second block is a polyol, such as poly(ethylene glycol), where the number of repeating ethylene glycol units is between 1 and 20. Suitable polymers for the second block may include, but are not limited to, another polyester copolymer of polyol and polycarboxylic acid, a hydrophilic polymer, hyaluronic acid, a polysaccharide, PEI, polylysine, polyglycolic acid, polylactic acid, or polycaprolactone.

[0056]

[0070] In some embodiments, the block copolymer comprises three or more blocks, such as a three-block or four-block, For example, a triblock copolymer may comprise a cationic first block, a hydrophobic second block, and an anionic or less cationic third block.

[0057] Polymer Delivery Complex

[0071] In an exemplary embodiment, the polymeric nanoparticle platform for delivery of a range of biologics and therapeutics is customizable to accommodate the needs of various payloads, including, but not limited to, nucleic acids, proteins, active pharmaceutical ingredients (APIs), gene editing systems, antibodies, cytokines, and other biologics. Polyester copolymers of polyols and polycarboxylic acids serve as the basic building blocks that compose the shape factors. Depending on the structure of the polymer and biologic, the shape factors may be polyplexes, micelles, polymersomes, niosomes, or polymer-lipid nanoparticles.

[0058]

[0072] In some embodiments, the nanoparticles are used for ex vivo delivery to cells.

[0059]

[0073] In some embodiments, the nanoparticles are used for in vivo delivery to cells.

[0060]

[0074] In some embodiments, the nanoparticles are used for in vitro delivery to cells.

[0061]

[0075] In some embodiments, the polymeric delivery system provides polymer structures that can be designed to fit a range of payload sizes for targeted delivery, tailored for specific payload release rates, and tuned to create a range of nanoparticle structures that can be decorated with specific functional moieties.

[0062]

[0076] If the polymer is a charged polymer, the charged polymer forms a polyplex when combined with a biologic payload molecule of opposite charge and mixed, where the polyplexes are linked by their opposite charges. In an exemplary embodiment, the polyplex is a nanoparticle.

[0063]

[0077] In some embodiments, polyester copolymers of polyols and polycarboxylic acids are modified with cationic moieties to enable the resulting charged polymer to bind negatively charged molecules, such as nucleic acids.

[0064]

[0078] In some embodiments, polyester copolymers of polyols and polycarboxylic acids are modified with anionic moieties to allow the charged polymer to bind positively charged molecules, such as certain peptides or proteins.

[0065]

[0079] In some embodiments, the polymeric delivery system incorporates the advantages of both polyplexes and nanoparticles in one form factor while maintaining non-immunogenicity. A polyester copolymer of polyol and polycarboxylic acid serves as a base structure to which various pendant building blocks can be added to achieve specific functions.

[0066]

[0080] Process levers for tuning the performance of polymeric nanoparticles can include, but are not limited to, molecular weight, charge density, acid:alcohol ratio of charged polymer building blocks, crosslink density, ratio of payload (biologic) to charged polymer, charged moieties used, and choice and presence of additives.

[0067]

[0081] In some embodiments, the polymeric delivery system includes an additive selected to adjust at least one property of the system. Suitable properties to be modified by the additive may include, but are not limited to, a number-weighted average particle size, zeta potential, water stability, cryostability, microbial resistance, plasma stability, or affinity for cell membranes. Suitable additives may include, but are not limited to, salts, buffers, surfactants, stabilizers, processing aids for cryopreservation, antimicrobial agents, or adjuvants. In some embodiments, the additive adjusts at least one property of the charged polymer. In some embodiments, the additive adjusts at least one property of the biologic complexed with the charged polymer. In some embodiments, the additive is an addendum that is inserted or assembled into the polymer-biologic complex, such as, for example, cholesterol, lipids, proteins, phospholipids, or synthetic amphiphilic molecules.

[0068]

[0082] Where the polyol includes glycerol and the polycarboxylic acid includes sebacic acid, suitable cationic moieties to bind to the PGS and maintain non-immunogenicity can include, but are not limited to, amines, ammonium, amino acids, peptides, peptide sequences, e.g., arginine-glycine-aspartic acid (RGD) to promote cell adhesion, choline, phosphocholine, sodium ions, potassium ions, or calcium ions.

[0069]

[0083] Suitable anionic moieties that bind to PGS and maintain it non-immunogenic may include, but are not limited to, sulfate, phosphate, sulfonate, sulfite, carboxylate, carbohydrate, or glycoprotein.

[0070]

[0084] As previously mentioned, PEG is often used to cloak lipid nanoparticles from the innate immune system, but PEG can cause allergic reactions and PEGylation is an additional step in nanoparticle production. Due to its non-immunogenicity, PGS, in exemplary embodiments, can reduce or eliminate the need for PEG or another cloaking material.

[0071]

[0085] In an exemplary embodiment, a charged polymer is mixed with an oppositely charged payload biologic molecule to form a polyplex and delivered to a cell. Once delivered to the cell, the payload molecule is released from the polyplex and the charged polymer then biodegrades with minimal toxicity to the cell.

[0072]

[0086] In some embodiments, the ratio of polymer to biologic is selected to aid in tailoring the average size and / or zeta potential of the resulting polymeric nanoparticles. In some embodiments, the ratio of polymer to biologic is selected to provide nanoparticles having a number average particle size in the range of about 150 nm to about 400 nm, or in the range of about 200 nm to about 300 nm, or any value, range, or subrange therebetween. In some embodiments, the ratio of polymer to biologic is selected to provide nanoparticles having a zeta potential in the range of about +10 mV to about +25 mV, or in the range of about +14 to about +22 mV, or any value, range, or subrange therebetween.

[0073]

[0087] In some embodiments, the charged polymer-biologic complexes may be multi-layered and assembled layer-by-layer, with alternating layers of positively and negatively charged species, hi some embodiments, the complexes may be capped with PEG to form a corona of hydration.

[0074]

[0088] In an exemplary embodiment, the polyplex delivery system is based on a building block approach to particle delivery. Figures 1 and 2 are schematic diagrams of the modular formation of the polyplex delivery system. Depending on the properties of the package, which may include, but are not limited to, size or charge, various complex formation techniques may be utilized. When the PGS is a polyester copolymer of polyol and polycarboxylic acid, the modular approach to polyplex generation preferably maintains the non-immunogenicity of the PGS.

[0075]

[0089] Referring to FIG. 2, the charged polymer 14 is mixed with a biologic 16 that naturally has an opposite net charge and assembles into a polyplex 18 based on the opposite charge. The formed polyplex 18 can then be used as a PGS-based polyplex delivery system. In some embodiments, the polyplex 18 is a nanoparticle. In some embodiments, the polyester copolymer 10 of polyol and polycarboxylic acid is modified with cationic moieties and the biologic 16 naturally has a net negative charge, which is shown diagrammatically as a nucleic acid in FIG. 2. In other embodiments, the polyester copolymer 10 of polyol and polycarboxylic acid is modified with anionic moieties and the biologic 16 naturally has a net positive charge.

[0076]

[0090] In some embodiments, the method for forming a delivery system includes combining a plurality of charged polymers and a plurality of biologics in a solvent to electrostatically bind the plurality of charged polymers and the plurality of biologics to generate a plurality of polyplexes, each polyplex of the plurality of polyplexes including at least one of the plurality of charged polymers and at least one of the plurality of biologics. Each of the plurality of charged polymers includes a polyester copolymer of a polyol and a polycarboxylic acid modified with at least one charged moiety having a net charge opposite to the net charge of the plurality of biologics. In some embodiments, the combining includes mixing. In some embodiments, the combining includes sonication. In some embodiments, the combining includes microfluidic mixing. In some embodiments, the solvent is an aqueous solvent. In some embodiments, the aqueous solvent is a PH buffered aqueous solvent. In some embodiments, the solvent is a co-solvent composition of ethanol and water. In some embodiments, the solvent includes dimethyl sulfoxide (DMSO).

[0077]

[0091] In some embodiments, the charged polymers and / or polyplexes contain a high charge density or high valency to improve the stability of the charged polymers and / or polyplexes prior to use and are stored in a solution, hi some embodiments, the charged polymers and / or polyplexes are lyophilized prior to use and stored in a low moisture environment.

[0078]

[0092] When the polymer is a block copolymer, C in formula (1) may be a polymer block selected to have an opposite charge to the biologic so as to form self-assembled particles of polyplexes by electrostatic complexation.

[0079]

[0093] Suitable sizes for the polyplexes in the polyplex delivery system range from 1 nanometer (nm) to 1000 nm, alternatively from about 10 nm to about 1000 nm, alternatively from about 50 nm to about 1000 nm, alternatively from about 100 nm to about 900 nm, alternatively from about 200 nm to about 800 nm, alternatively from about 100 nm to about 500 nm, alternatively from about 50 nm to about 100 nm, alternatively from about 50 nm to about 200 nm, alternatively from about 50 nm to about 500 nm, alternatively from about 50 nm to about 800 nm, alternatively from about 100 nm to about 200 nm, alternatively from about 300 nm to about 500 nm, alternatively from about 500 nm to about 900 nm, or any value, range, or subrange therebetween. Although the particles are generally described herein as nanoparticles, the polyplex number weighted average particle size may extend into the microparticle size range in some embodiments.

[0080]

[0094] Although the polyplex nanoparticle delivery system has been described primarily for delivering anionic nucleic acids as biologics, the polymeric delivery system can be modified to deliver other anionic or cationic biologics. Other suitable charged biologics can include, but are not limited to, enzymes, proteins, APIs, peptides, amino acids, gene editing systems, antibodies, cytokines, or aptamers.

[0081]

[0095] The cationic character of the charged polymers described herein and shown in FIG. 1 may be replaced by anionic character depending on the charge of the biologic.

[0082]

[0096] In some embodiments, a method for delivering a biologic to a cell comprises administering a polyplex of the biologic electrostatically associated with a charged polymer, the charged polymer comprising a polyester copolymer of a polyol and a polycarboxylic acid modified with at least one charged moiety having an opposite charge to the net charge of the biologic. In some embodiments, the polyplex is administered parenterally.

[0083]

[0097] In some embodiments, the values ​​of a, b, c, d, x, and z in formula (1) are selected such that the block copolymer, alone or in combination with a biologic, self-assembles to form a polymersome.

[0084]

[0098] In some embodiments, the values ​​of a, b, c, d, x, and z in formula (1) are selected such that the block copolymer, alone or in combination with a biologic, forms self-assembled particles of polymeric micelles.

[0085]

[0099] In some embodiments, the self-assembled particles are nanoparticles. Suitable sizes for the self-assembled particles of the polymeric delivery system are in the range of 1 nanometer (nm) to 1000 nm, alternatively about 50 nm to about 1000 nm, alternatively about 100 nm to about 900 nm, alternatively about 200 nm to about 800 nm, alternatively about 100 nm to about 500 nm, alternatively about 50 nm to about 100 nm, alternatively about 50 nm to about 200 nm, alternatively about 50 nm to about 500 nm, alternatively about 50 nm to about 800 nm, alternatively about 100 nm to about 200 nm, alternatively about 300 nm to about 500 nm, alternatively about 500 nm to about 900 nm, or any value, range, or subrange therebetween. Although the particles are generally described herein as nanoparticles, the particle size may extend into the microparticle size range in some embodiments.

[0086]

[0100] In an exemplary embodiment, the polymeric delivery system comprising block copolymers is based on a building block approach to particle delivery. Figures 3 and 4 are schematic diagrams of two molecular formulations of polymeric delivery systems derived from block copolymers and biologics. In Figure 3, the block copolymer and biologic self-assemble into polymeric micelles. In Figure 4, the block copolymer and biologic self-assemble into polymersomes. Instead of the attached hydrophobic chains shown in Figures 3 and 4, the block copolymer may comprise a hydrophobic first block polymer or a hydrophobic second block polymer. Depending on the properties of the package, which may include, but are not limited to, size, hydrophobicity, or charge, various particle technologies may be utilized. The modular approach to particle generation not only maintains the non-immunogenicity of the constituent monomeric polymers, but also the bioabsorbability of the first block.

[0087]

[0101] Referring to formula (1), the alcohol of the AB first block of the polyester copolymer of polyol and polycarboxylic acid belonging to the block copolymer is not derivatized and C is used to impart functionality to the delivery system, for example, charge, hydrophilicity, or molecular weight.

[0088]

[0102] In some embodiments, the alcohol groups of the first block are used as tether moieties that can be derivatized to impart additional functionality, where the alcohol is converted to an alkyl tail to impart amphiphilic properties to the delivery system, which allows for the formation of polymeric micelle structures, as shown generally in FIG. 3, and polymersome structures, as shown generally in FIG. 4. The alkyl tails can be single or double tails. The alkyl tails can include any suitable degree of unsaturation, including saturation. The alkyl tails can be, for example, C5 to C6 alkyl tails, including any integer value, range, or subrange between 5 and 22. 22In some embodiments, the anionic biologic is a single- or double-stranded nucleic acid, which may be DNA or RNA.

[0089]

[0103] In an exemplary embodiment, the alcohol groups of the first block are derivatized with a charged molecule or polymer to enhance the overall charge characteristics of the delivery system.

[0090]

[0104] In some embodiments, the alcohol of the first block is derivatized with PEG to increase the hydrophilicity of the delivery system and provide an aqueous corona around the nanoparticle.

[0091]

[0105] It will be appreciated by those skilled in the art that derivatization of the alcohol groups of the first block creates degradable linkages, allowing for degradation of the polymeric delivery system upon payload delivery.

[0092]

[0106] In some embodiments, the method for forming a polymeric delivery system includes combining a block copolymer and a biologic in a solvent to form a self-assembled particle that includes the block copolymer and the biologic bound to the block copolymer. In some embodiments, the combining includes mixing. In some embodiments, the combining includes sonication. In some embodiments, the combining includes microfluidic mixing. In some embodiments, the solvent is an aqueous solvent. In some embodiments, the aqueous solvent is a PH buffered aqueous solvent. In some embodiments, the solvent is a co-solvent composition of ethanol and water. In some embodiments, the solvent includes DMSO.

[0093]

[0107] In some embodiments, the polymeric delivery system formed is in the form of a polymeric micelle. Figure 3 shows some alcohol hydrophobic moieties of the first and second blocks of a block copolymer that contain charged groups that form electrostatic interactions with oppositely charged biologics. The block copolymer and biologics self-assemble in aqueous media into a micelle-like structure with a hydrophobic core.

[0094]

[0108] In some embodiments, the polymeric delivery system formed is in the form of a polymersome. Figure 4 shows a hydrophobic moiety grafted from some alcohol groups of the first block and second block of one of the block copolymers containing a charged group that forms an electrostatic interaction with the oppositely charged biologic. The block copolymer and the biologic self-assemble in the solvent into a polymersome with a bilayer of hydrophobic moieties and a core that includes the charged second block complexed with the biologic.

[0095]

[0109] In some embodiments, the above-mentioned micelles or polymersomes are designed to be used with conventional lipid nanoparticle technology to create hybrid nanoparticles. Conventional lipid nanoparticles are composed of amphiphilic lipids with charged head groups attached to an oppositely charged core. The resulting structure exposes the hydrophobic tails on the outside of the particle. The amphiphilic block copolymers described herein can be oriented so that the hydrophobic tails of the polymer conjugate are in the same direction as the hydrophobic tails of the lipid nanoparticle. The addition of polymeric components to the hybrid particles stabilizes the LNP core and protects it from dissociation, opsonizing proteins, or innate or adaptive immune responses. Additionally, the polymers may be derivatized with targeting ligands to create hybrid nanoparticles that can deliver payloads to specific biological targets.

[0096]

[0110] In some embodiments, the polymeric delivery systems are designed to incorporate specific segments that can interact with portions of the biologic payload to produce amorphous solid dispersions, condense the payload to produce smaller particles, enhance the release rate, stabilize the biologic, protect the biologic from opsonization or other immune system attack, improve plasma stability to increase circulation time, or provide other benefits.

[0097]

[0111] In some embodiments, a hybrid particle system is created in which the polymer delivery system acts as a condensing agent to generate smaller payload particles, which can be further processed using standard lipid nanoparticle formation methods.

[0098]

[0112] In some embodiments, the block copolymers are lyophilized prior to use and stored in a maintained low moisture environment.

[0099]

[0113] Although the polymeric delivery systems have been described primarily for delivering anionic nucleic acids as biologics, the polymeric delivery systems can be modified to deliver cationic or uncharged biologics, including, but not limited to, enzymes, proteins, APIs, peptides, amino acids, or aptamers.

[0100]

[0114] The cationic character described herein and shown in Figures 3 and 4 may be replaced with anionic, neutral, hydrophobic, or hydrophilic moieties depending on the binding mechanism between the block copolymer and the biologic.

[0101]

[0115] In some embodiments, the biologic is polar or hydrophilic, or comprises a polar or hydrophilic region that binds to a polar or hydrophilic portion or region of the block copolymer.

[0102]

[0116] In some embodiments, the biologic is non-polar or hydrophobic, or includes a non-polar or hydrophobic region that associates with a non-polar or hydrophobic portion or region of the block copolymer.

[0103]

[0117] In some embodiments, a method for delivering a biologic to a cell comprises administering a self-assembled particle comprising a block copolymer and a biologic bound to the block copolymer, hi some embodiments, the self-assembled particle is administered parenterally.

[0104]

[0118] Other applications of the polymer delivery system include, but are not limited to, the formation of chimeric antigen receptor T cells (CAR-T cells); treatment of solid tumors or blood cancers; other cell / gene-based therapies, such as, but not limited to, tumor infiltrating lymphocyte (TIL) therapy, artificial T cell receptor (TCR) therapy, natural killer (NK) therapy, ocular cell therapy, neural cell therapy, skin cell therapy, beta cell therapy, pluripotent stem cell therapy, adult stem cell therapy, cancer stem cell therapy, dendritic cell therapy, lymphokine-activated killer (LAK) cell therapy, cytokine-induced killer (CIK) cell therapy, gamma delta T cell therapy, and the like. cell therapy, umbilical cord blood (CB) cell therapy, hematopoietic stem cell (HSC) therapy, mesenchymal stem cell (MSC) therapy; enzyme encapsulation for bioremediation of plastics; controlled release of fertilizers, growth promoters, pesticides, or herbicides in agricultural environments; controlled delivery of substances that respond to external stimuli, such as nanoparticles that degrade when exposed to a particular pH range and release an encapsulated dye for colorimetric detection, or nanoparticles that degrade when subjected to mechanical stress or shear and release a payload for internal sensing in rubber belts or tires; or transdermal delivery.

[0105]

[0119] For example, in some embodiments where the polymeric delivery system provides a solid tumor cancer treatment, the charged polymer of the polyplex comprises glutamine. In some embodiments, glutamine is incorporated into the repeat unit of the charged polymer as an additional monomer to the polyol and polycarboxylic acid in the polymerization step. In some embodiments, glutamine is used to modify the formed polyester copolymer of polyol and polycarboxylic acid to provide a pendant charged moiety to the charged polymer.

[0106]

[0120] In some embodiments, the polyplexes described herein are coated on one or more surfaces of a textile graft or other implanted tissue to help induce blood clotting at the surface of the implanted tissue. For example, in aneurysm repair with textile grafts, blood often leaks through suture hole vias, which causes delayed clot formation and transient thrombosis. Polyplex coating induces clotting on the textile graft surface by attracting platelets to the coated surface, which induces blood clotting, thereby sealing the holes and preventing blood leakage from the graft. EXAMPLES

[0107]

[0121] The present invention will be further described with reference to the following examples, which are offered by way of illustration and not by way of limitation.

[0108] Example 1

[0122] Poly(glycerol sebacate) was synthesized by a water-mediated method described in U.S. Patent No. 9,359,472 issued on June 7, 2016. Then, 15 g of poly(glycerol sebacate) was placed in a reactor and 15 g of dimethylaminopropanoic acid was added. The mixture was then heated and maintained at 120° C. for 24 hours under nitrogen flow. The resulting mixture was purified by reversed-phase column chromatography to obtain a charged polymer of cationic modified poly(glycerol sebacate) polymer. Gel permeation chromatography (GPC) results showed that the weight average molecular weight of the resulting charged polymer was 4500 Da and the polydispersity index (D) was 3.0. Dynamic light scattering (DLS) results showed that the average particle size was 78 nm and the zeta potential was +20 mV.

[0109] Example 2

[0123] Poly((glycerol sebacate 10% Succinate 90%Poly((glycerol sebacate 10% succinate 90%)-co-xylitol) was synthesized using a water-mediated method. Glycerol (100.0 g), sebacic acid (22.0 g), succinic acid (115.4 g), and water (25.4 g) were added to a reactor. The mixture was then melted and maintained at a temperature of 120° C. for 48 hours under nitrogen flow and mechanical stirring. Xylitol (26.0 g) was then added to the reaction mixture and stirred for 24 hours. GPC results showed that the weight average molecular weight of the resulting polymer was 4500 Da (D4.8). Figure 5A shows the APT of poly((glycerol sebacate 10% succinate 90%)-co-xylitol). 13 1 is a graph showing a C-NMR spectrum.

[0110]

[0124] 15 g of poly((glycerol sebacate) 10% Succinate 90% Poly((glycerol sebacate)) was reacted with dimethylaminopropanoic acid (15 g) containing p-toluenesulfonic acid as a catalyst in the melt at 130 °C for 24 h with mechanical stirring to give poly((glycerol sebacate)). 10% Succinate 90% The resulting mixture was then placed in a dialysis tube and dialyzed for 4 days, after which it was freeze-dried. The GPC results showed that the weight-average molecular weight of the resulting polymer was 9800 Da (D1.9). Figure 5B shows the APT of poly((glycerol sebacate 10% succinate 90%)-co-xylitol dimethylaminopropylamine). 13 1 is a graph showing a C-NMR spectrum. The peaks at 53 ppm and 42 ppm (arrows) represent tertiary amines after addition of dimethylaminopropanoic acid.

[0111]

[0125] FIG. 6 is a graph showing the FTIR-ATR spectra of poly((glycerol sebacate 10% succinate 90%)-co-xylitol) (dashed line) and poly((glycerol sebacate 10% succinate 90%)-co-xylitol dimethylaminopropylamine) (solid line). -1The absorbance at 3400 cm (arrow) represents the tertiary amine after addition of dimethylaminopropanoic acid. -1 The decrease in -OH absorbance at 100° C. further indicates reaction with the -OH groups of the polyol monomer.

[0112] Example 3

[0126] Poly((glycerol sebacate) of Example 2 10% Succinate 90% )-co-xylitol dimethylaminopropylamine) was passed through a size exclusion column to obtain a low dispersion polymer (Mw4500D2). This polymer was then reacted with carbonyldiimidazole and stirred for 3 hours. 1,1-dimethylethylenediamine was then added and stirred for 12 hours. The resulting mixture was then placed in a dialysis tube and dialyzed for 4 days, after which it was freeze-dried. GPC results showed that the weight average molecular weight of the resulting polymer was 1546Da (D1.1).

[0113] Example 4

[0127] Poly((PEG300) 10% Glycerol 90% Succinate-co-xylitol was synthesized using a water-mediated method. Glycerol (71.7 g), polyethylene glycol 300 (25.9 g), succinic acid (175 g), and water (20.2 g) were added to a reactor and heated to 120° C. under nitrogen flow with mechanical stirring for 48 hours. Xylitol (27.3 g) was then added and the reaction was carried out for another 24 hours. GPC results showed that the weight average molecular weight of the resulting polymer was 5300 Da (D5.3).

[0114]

[0128] 15g of Poly(PEG300) 10% Glycerol 90%Succinate-co-xylitol (15 g) was reacted with dimethylaminopropanoic acid (15 g) containing p-toluenesulfonic acid as catalyst in the melt at 130 °C for 24 h under mechanical stirring. The resulting mixture was then placed in a dialysis tube and dialyzed for 4 days, followed by freeze-drying. GPC results showed that the weight average molecular weight of the resulting polymer was 10,800 Da (D 1.20). FTIR showed a peak at 1586 cm -1 , indicating functionalization of the polymer.

[0115] Example 5

[0129] Poly((glycerol succinate)-co-xylitol) was synthesized using a water-mediated method. Glycerol (100.0 g), succinic acid (128.0 g), and water (25.4 g) were added to a reactor. The mixture was then melted and maintained at a temperature of 120° C. for 24 hours under nitrogen flow and mechanical stirring. Xylitol (22.8 g) was then added to the reaction mixture and stirred for 24 hours. GPC results showed that the weight average molecular weight of the resulting polymer was 2724 Da (D3.6).

[0116]

[0130] 15 g of poly((glycerol succinate)-co-xylitol) was reacted with dimethylaminopropanoic acid (15 g) containing 1% p-toluenesulfonic acid as catalyst at 130 °C in the melt with mechanical stirring for 24 h. The resulting mixture was then placed in a dialysis tube and dialyzed for 4 days, followed by freeze-drying. GPC results showed that the weight average molecular weight of the resulting polymer was 20,200 Da (D 1.52). FTIR showed a peak at 1586 cm -1 , indicating functionalization of the polymer.

[0117] Example 6

[0131] Poly(((PEG300) 10% Glycerol 90%Glycerol (94 g), polyethylene glycol 300 (34.1 g) (PEG300), glutaric acid (150 g), and water (26.6 g) were added to a reactor. The mixture was then melted and maintained at a temperature of 120° C. under nitrogen flow and mechanical stirring for 48 hours. Xylitol (30.5 g) was then added to the reaction mixture and stirred for 24 hours. GPC results showed that the weight average molecular weight of the resulting polymer was 8280 Da (D8.6).

[0118]

[0132] 15g of Poly((PEG300) 10% Glycerol 90% (Glutarate)-co-xylitol) was reacted with dimethylaminopropanoic acid (15 g) containing 1% p-toluenesulfonic acid as catalyst in the melt at 130 °C for 24 h under mechanical stirring. The resulting mixture was then placed in a dialysis tube and dialyzed for 4 days, followed by freeze-drying. GPC results showed that the weight average molecular weight of the resulting polymer was 24,980 Da (D1.50). FTIR showed a peak at 1586 cm -1 , indicating functionalization of the polymer.

[0119] Example 7

[0133] Poly(((TPEG1000) 50% Glycerol 50% Succinate-co-xylitol) was synthesized using a water-mediated method. Glycerol (19.49 g), trimethylolpropane ethoxylate 1000 (214.83 g) (TPEG1000), succinic acid (50.0 g), and water (9.9 g) were added to a reactor. The mixture was then melted and maintained at a temperature of 120° C. under nitrogen flow and mechanical stirring for 48 hours. Xylitol (29.4 g) was then added to the reaction mixture and stirred for 24 hours. GPC results showed that the weight average molecular weight of the resulting polymer was 4850 Da (D4.1).

[0120]

[0134] 15g of Poly((TPEG1000) 50% Glycerol50% Succinate-co-xylitol was reacted with dimethylaminopropanoic acid (15 g) containing 1% p-toluenesulfonic acid as catalyst at 130 °C in the melt for 24 hours under mechanical stirring. The resulting mixture was then placed in a dialysis tube and dialyzed for 4 days, followed by freeze-drying. GPC results showed that the weight average molecular weight of the resulting polymer was 4,270 Da (D-1.52). FTIR showed a peak at 1586 cm -1 , indicating functionalization of the polymer.

[0121] Example 8

[0135] Poly(((PEG600) 25% Glycerol 75% Succinate-co-xylitol) was synthesized using a water-mediated method. Glycerol (73.1 g), polyethylene glycol 600 (158.9 g) (PEG600), succinic acid (125.0 g), and water (24.8 g) were added to a reactor. The mixture was then melted and maintained at a temperature of 120° C. under nitrogen flow and mechanical stirring for 48 hours. Xylitol (38.18 g) was then added to the reaction mixture and stirred for 24 hours. GPC results showed that the weight average molecular weight of the resulting polymer was 3380 Da (D3.4).

[0122]

[0136] 15g of Poly((PEG600) 25% Glycerol 75% Succinate-co-xylitol was reacted with dimethylaminopropanoic acid (15 g) containing 1% p-toluenesulfonic acid as catalyst at 130 °C in the melt for 24 hours under mechanical stirring. The resulting mixture was then placed in a dialysis tube and dialyzed for 4 days, followed by freeze-drying. GPC results showed that the weight average molecular weight of the resulting polymer was 2900 (D 1.7). FTIR showed a peak at 1586 cm -1 , indicating functionalization of the polymer.

[0123] Example 9

[0137] Poly(((PEG600) 10% Glycerol 90%Succinate-co-xylitol) was synthesized using a water-mediated method. Glycerol (87.7 g), polyethylene glycol 600 (63.6 g), succinic acid (125.0 g), and water (24.8 g) were added to a reactor. The mixture was then melted and maintained at a temperature of 120° C. for 48 hours under nitrogen flow and mechanical stirring. Xylitol (30.11 g) was then added to the reaction mixture and stirred for 24 hours. GPC results showed that the weight average molecular weight of the resulting polymer was 4895 Da (D4.8).

[0124]

[0138] 15g of Poly((PEG600) 10% Glycerol 90% Succinate-co-xylitol was reacted with dimethylaminopropanoic acid (15 g) containing 1% p-toluenesulfonic acid as catalyst at 130 °C in the melt for 24 hours under mechanical stirring. The resulting mixture was then placed in a dialysis tube and dialyzed for 4 days, followed by freeze-drying. GPC results showed that the weight average molecular weight of the resulting polymer was 5210 (D1.50). FTIR showed a peak at 1586 cm -1 , indicating functionalization of the polymer.

[0125] Example 10

[0139] Poly((glycerol glutarate)-co-xylitol) was synthesized using a water-mediated method. Glycerol (100.0 g), glutaric acid (143.0 g), and water (25.4 g) were added to a reactor. The mixture was then melted and maintained at a temperature of 120° C. under nitrogen flow and mechanical stirring for 48 hours. Xylitol (26.8 g) was then added to the reaction mixture and stirred for 24 hours. GPC results showed that the weight average molecular weight of the resulting polymer was 4986 Da (D5.5).

[0126]

[0140] 15 g of poly((glycerol glutarate)-co-xylitol) was reacted with dimethylaminopropanoic acid (15 g) containing 1% p-toluenesulfonic acid as catalyst at 130 °C in the melt with mechanical stirring for 24 h. The resulting mixture was then placed in a dialysis tube and dialyzed for 4 days, followed by freeze-drying. GPC results showed that the weight average molecular weight of the resulting polymer was 25,100 Da (D 1.38). FTIR showed a peak at 1586 cm -1 , indicating functionalization of the polymer.

[0127] Example 11

[0141] Poly((glycerol glutarate) 50% Succinate 50% )-co-xylitol) was synthesized using a water-mediated method. Glycerol (100.0 g), glutaric acid (71.7 g), succinic acid (64.1 g), and water (25.4 g) were added to a reactor. The mixture was then melted and maintained at a temperature of 120° C. for 48 hours under nitrogen flow and mechanical stirring. Xylitol (26.0 g) was then added to the reaction mixture and stirred for 24 hours. GPC results showed that the weight average molecular weight of the resulting polymer was 5141 Da (D5.5).

[0128]

[0142] 15 g of poly((glycerol glutarate) 50% Succinate 50% )-co-xylitol) was reacted with dimethylaminopropanoic acid (15 g) containing 1% p-toluenesulfonic acid as catalyst at 130 °C in the melt with mechanical stirring for 24 h. The resulting mixture was then placed in a dialysis tube and dialyzed for 4 days, followed by freeze-drying. GPC results showed that the weight average molecular weight of the resulting polymer was 7500 Da (D 1.58). FTIR showed a peak at 1586 cm -1 , indicating functionalization of the polymer.

[0129] Example 12

[0143] Poly((glycerol adipate)-co-xylitol) was synthesized using a water-mediated method. Glycerol (125.0 g), adipic acid (198.0 g), and water (32.0 g) were added to a reactor. The mixture was then melted and maintained at a temperature of 120° C. under nitrogen flow and mechanical stirring for 48 hours. Xylitol (35.5 g) was then added to the reaction mixture and stirred for 24 hours. GPC results showed that the weight average molecular weight of the resulting polymer was 9842 Da (D10.5).

[0130]

[0144] 15 g of poly((glycerol adipate)-co-xylitol) was reacted with dimethylaminopropanoic acid (15 g) containing 1% p-toluenesulfonic acid as a catalyst in the melt at 130° C. for 24 h with mechanical stirring. The resulting mixture was then placed in a dialysis tubing and dialyzed for 4 days before being freeze-dried.

[0131] Example 13

[0145] Poly((glycerol adipate 50% Succinate 50% )-co-xylitol) was synthesized using a water-mediated method. Glycerol (113.0 g), adipic acid (90.0 g), succinic acid (73.0 g), and water (14.0 g) were added to a reactor. The mixture was then melted and maintained at a temperature of 120° C. for 48 hours under nitrogen flow and mechanical stirring. Xylitol (29.0 g) was then added to the reaction mixture and stirred for 24 hours. GPC results showed that the weight average molecular weight of the resulting polymer was 7037 Da (D7.1).

[0132]

[0146] 15 g of poly((glycerol adipate) 50% Succinate 50%)-co-xylitol) was reacted with dimethylaminopropanoic acid (15 g) containing 1% p-toluenesulfonic acid as catalyst at 130 °C in the melt with mechanical stirring for 24 h. The resulting mixture was then placed in a dialysis tube and dialyzed for 4 days, followed by freeze-drying. GPC results showed that the weight average molecular weight of the resulting polymer was 9500 Da (D 1.56). FTIR showed a peak at 1586 cm -1 , indicating functionalization of the polymer.

[0133] Example 14

[0147] Poly((glycerol 50% Xylitol 50% ) succinate) was synthesized using a water-mediated method. Glycerol (19.5 g), xylitol (32.2 g), succinic acid (50.0 g), and water (9.9 g) were added to a reactor. The mixture was then melted and maintained at a temperature of 120° C. for 24 hours under nitrogen flow and mechanical stirring. GPC results showed that the weight average molecular weight of the resulting polymer was 1219 Da (D 1.9).

[0134]

[0148] 15 g of poly((glycerol 50% Xylitol 50% ) succinate) was reacted with dimethylaminopropanoic acid (15 g) containing 1% p-toluenesulfonic acid as a catalyst in the melt with mechanical stirring at 130 °C for 24 h. The resulting mixture was then placed in dialysis tubing and dialyzed for 4 days, after which it was lyophilized.

[0135] Example 15

[0149] Poly(glycerol succinate) was synthesized using a water-mediated method. Glycerol (100.0 g), succinic acid (128.0 g), and water (25.4 g) were added to a reactor. The mixture was then melted and maintained at a temperature of 120° C. for 48 hours under nitrogen flow and mechanical stirring. GPC results showed that the weight average molecular weight of the resulting polymer was 3,064 Da (D3.2).

[0136]

[0150] 15 g of poly(glycerol succinate) was reacted with dimethylaminopropanoic acid (15 g) containing 1% p-toluenesulfonic acid as a catalyst in the melt with mechanical stirring at 130° C. for 24 h. The resulting mixture was then placed in dialysis tubing and dialyzed for 4 days before being lyophilized.

[0137] Example 16

[0151] Poly(glycerol N-methyliminodiacetate) was synthesized by adding glycerol (3 g) to a round-bottom flask equipped with a magnetic stirrer and heating to 120 °C under nitrogen flow. N-methyliminodiacetic acid (4.8 g) was added slowly until a homogeneous mixture was observed. The reactants were then stirred at 120 °C for 4 h. The reaction mixture was then dissolved in deionized water and dialyzed. GPC results showed that the weight average molecular weight of the resulting polymer was 3,000 Da (D 1.6).

[0138] Example 17

[0152] The cationic modified PGS charged polymer of Example 1 was mixed with mRNA containing a GFP reporter (Dasher GFP mRNA, Aldevron, Fargo, ND) at weight ratios of mRNA to charged polymer of 1:1, 1:2, 1:4, and 1:16. The mixture was shaken on an orbital shaker at 100 rpm for 20 minutes to form polyplex particles ranging in size from nanometers to micrometers when viewed under a microscope.

[0139] Example 18

[0153] The polyplex particles of Example 17, in which the weight ratio of mRNA:charged polymer was 1:2, were introduced into the cells, and 7.38 × 10 4 A total of 500ng of mRNA was present per well in a 24-well plate containing human cardiac fibroblasts seeded at a density of 100x100 cells. The polyplex particles were incorporated into the cells, and GFP reporter fluorescence showed that the cells were successfully transfected after 21 hours. The transfected cells were visible under a fluorescent microscope and a light microscope.

[0140] Example 19

[0154] Poly((glycerol sebacate) of Example 2 10% Succinate 90% Polyplex particles containing 0.25 μg of Dasher mRNA (Aldevron) were prepared using PGSSuX 1-9D) to express a GFP reporter in human cardiac fibroblasts. 10 μL of polyplex particles prepared at a weight ratio of 100:1 PGSSuX 1-9D:mRNA were added to cells cultured in 96-well microplates containing 12,500 cells per well. Cell transfection was monitored over 96 hours with a Celllink CELLCYTE X (Cytena GmbH, Freiburg, Germany) live cell imaging system, with images of cells taken once per hour during transfection. Images taken 3 hours after transfection showed no transfection at early time points. Images taken 5 hours after transfection showed visible evidence of some successful transfection. An increased number of cells showed fluorescence upon transfection of mRNA at later time points, for example 18 hours post-transfection.

[0141] Example 20

[0155] Poly((glycerol sebacate) of Example 2 10% Succinate 90%Polyplex particles containing 12.6 μg of gWiZ pDNA (Aldevron) with a size of 5757 base pairs were prepared using PGS-co-xylitol dimethylaminopropylamine (PGSSuX 1-9D) to express a GFP reporter in mouse NIH / 3T3 cells (American Type Tissue Culture, Manassas, VA). 10 μL of polyplex particles prepared at a weight ratio of PGSSuX 1-9D:pDNA of 140:1 were added to cells cultured in 96-well microplates containing 10,000 cells per well. Cell transfection was monitored over 96 hours with a Celllink CELLCYTE X (Cytena) live cell imaging system, with images of cells taken once per hour during transfection. 29 hours after addition of PGS polyplex particles, 10% of cells were transfected.

[0142] Example 21

[0156] Poly((glycerol sebacate) of Example 2 10% Succinate 90%Polyplex particles containing 20 pg of Dasher GFP mRNA (Aldevron) with a size of 755 nucleotides with a 250 nucleotide poly(A) tail were prepared using PGSSuX 1-9D, and delivered per cell to express the GFP reporter in primary human cardiac fibroblasts (PromoCell, Heidelberg, Germany). 10 μL of polyplex particles in 25 mM sodium acetate, prepared at a weight ratio of 100:1 PGSSuX 1-9D:mRNA, were added to cells cultured in 96-well microplates containing 12,500 cells per well. Cell transfection was monitored over a 120-h period with a Celllink CELLCYTE X live-cell imaging system, and images of cells were taken once per hour in bright field and green filter channels to monitor transfection. As shown in FIG. 7, 48 hours after addition of PGS polyplex particles, the cells showed peak levels of transfection.

[0143] Example 22

[0157] Poly((glycerol sebacate) of Example 2 10% Succinate 90%Polyplex particles containing 140 pg of gWiZ pDNA (Aldevron) with a size of 5757 base pairs were prepared using PGS-co-xylitol dimethylaminopropylamine (PGSSuX 1-9D) and delivered per cell to express the GFP reporter in mouse NIH / 3T3 cells (American Type Tissue Culture, Manassas, VA). 10 μL of polyplex particles in 25 mM sodium acetate, prepared at a weight ratio of PGSSuX 1-9D:pDNA of 100:3, were added to cells cultured in 96-well microplates containing 10,000 cells per well. Cell transfection was monitored over a 96-hour period with a Celllink CELLCYTE X live-cell imaging system, and images of cells were taken once per hour during transfection in bright field and green filter channels to monitor transfection. As shown in Figure 8, 48 hours after addition of PGS polyplex particles, cells showed peak levels of transfection.

[0144] Example 23

[0158] Poly((glycerol sebacate) of Example 2 10% Succinate 90%Polyplex particles containing 60 pg of gWiZ pDNA (Aldevron) with a size of 5757 base pairs were prepared using PGS-co-xylitol dimethylaminopropylamine (PGSSuX 1-9D) and delivered per cell to express the GFP reporter in mouse NIH / 3T3 cells (American Type Tissue Culture, Manassas, VA). 10 μL of polyplex particles in 25 mM sodium acetate, prepared at a weight ratio of PGSSuX 1-9D:pDNA of 50:2, were added to cells cultured in 96-well microplates containing 10,000 cells per well. Cell transfection was monitored over a 96-hour period with a Celllink CELLCYTE X live-cell imaging system, and images of cells were taken once per hour during transfection in bright field and green filter channels to monitor transfection. As shown in Figure 9, 48 hours after addition of PGS polyplex particles, cells showed peak levels of transfection.

[0145] Example 24

[0159] Polyplex particles containing 10 pg of gWiZ pDNA (Aldevron) with a size of 5757 base pairs were prepared using poly((glycerol glutarate)-co-xylitol) (PGGXD) from Example 10 and delivered per cell to express GFP reporter in Jurkat E6.1 cell line (American Type Tissue Culture, Manassas, VA). 20 μL of polyplex particles in 25 mM sodium acetate, prepared at a weight ratio of 25:1 PGGXD:pDNA, were added to cells cultured in 96-well microplates containing 250,000 cells per well. Cell transfection was monitored over 48 hours with a Celllink CELLCYTE X live cell imaging system, and images of cells were taken once every 3 hours during transfection in bright field and green filter channels to monitor transfection. As shown in FIG. 10, 24 hours after addition of PGS polyplex particles, cells showed peak levels of transfection.

[0146] Example 25

[0160] The polyplex particles of Example 17 with a weight ratio of mRNA:charged polymer of 1:40 were characterized for zeta potential and size with a Zetasizer Ultra (Malvern Panalytical Ltd., Malvern, UK) light scattering system. The results are shown in Figures 11 and 12. The polyplex particles were found to have an average particle size of 167.3 nm, a polydispersity of 0.068, and a slightly negative zeta potential of -7.929 mV due to complexation of the charged polymer with the mRNA.

[0147] Example 26

[0161] The polyplex particles of Example 20 with a weight ratio of pDNA:charged polymer of 1:50 were characterized for zeta potential and size with a Zetasizer Ultra (Malvern Panalytical Ltd., Malvern, UK) light scattering system. The results are shown in Figures 13 and 14. The polyplex particles were found to have an average particle size of 135 nm and a slightly positive zeta potential of 23.135 mV due to complexation of the charged polymer with pDNA.

[0148] Example 27

[0162] Equal molar ratios of glycerol (1.83 g) and succinic acid (2.34 g) were added to a 50 mL round bottom flask and heated to 130 °C with magnetic stirring. Once the mixture was homogenous, dry lysine (2.9 g) in a 1:1 molar ratio to glycerol was slowly added to the round bottom flask. Upon addition of lysine, gas was evolved. After gas evolution ceased, more lysine was added. After all lysine was added, a nitrogen flow was applied and the reaction was continued for 12 h. GPC results showed that the weight average molecular weight of the resulting poly(lysine glycerol succinate) polymer was 3430 Da (D 5.09).

[0149] Example 28

[0163] Diglycidyl sebacate (0.5 g, 0.159 mmol), 4,7,10-trioxa-1,13-tridecanediamine (348 μL, 0.159 mmol), tributylammonium bromide (8 mg, 0.025 mmol), and dimethylformamide (DMF) (1 mL) were added to a round-bottom flask equipped with a magnetic stirrer. The mixture was then heated to 70 °C for 3 h. The reaction was then removed from the heat and cooled to room temperature, resulting in a viscous liquid. Gel permeation chromatography (GPC) results showed that the weight average molecular weight of the resulting polymer was 1700 Da (D-1.8).

[0150] Example 29

[0164] A polyplex formulation consisting of 1 μg of the polymer of Example 27 mixed with 500 ng of green fluorescent protein (GFP) reporter mRNA (Dasher GFP mRNA, Aldevron, Fargo, ND) was added to 1.0 × 10 5 The cells were then added to a 24-well plate containing fibroblasts. After 24 hours, the cells were imaged under a fluorescent microscope to visualize the transfected cells.

[0151]

[0165] The polyplex particles were imaged under a microscope in bright field mode. The cells were imaged under a fluorescent microscope and a light microscope, and the fluorescence showed that the cells were successfully transfected.

[0152]

[0166] All of the above references are incorporated herein by reference.

[0153]

[0167] Although the present invention has been described with reference to one or more exemplary embodiments, those skilled in the art will recognize that various changes may be made and equivalents substituted for the elements without departing from the scope of the invention. In addition, many modifications may be made to adapt a particular situation or material to the teachings of the invention without departing from the essential scope of the invention. Therefore, the invention is not limited to the particular embodiment disclosed as the best mode contemplated for carrying out the invention, but the invention is intended to include all embodiments falling within the scope of the appended claims. Furthermore, all numerical values ​​identified in the detailed description should be construed as if both exact and approximate values ​​were explicitly identified.

Claims

1. A polymeric delivery system comprising nanoparticles, each nanoparticle comprising at least one polymer and at least one biological agent, wherein the at least one polymer comprises a polyester copolymer of a polyol and a polycarboxylic acid.

2. 10. The polymeric delivery system of claim 1, wherein the nanoparticles are polyplexes, the polymers are charged polymers, each polyplex comprises at least one charged polymer and at least one biological agent, and the at least one charged polymer comprises a polyester copolymer of a polyol and a polycarboxylic acid modified with at least one charged moiety having an opposite charge to the net charge of the at least one biological agent.

3. 3. The polymeric delivery system of claim 2, wherein the polyplexes have a number-weighted average particle size ranging from about 10 nm to about 1000 nm.

4. The polymeric delivery system of claim 2 further comprising a solvent.

5. 3. The polymeric delivery system of claim 2, wherein the polyester copolymer of polyol and polycarboxylic acid comprises a copolymer of glycerol and a polycarboxylic acid selected from the group consisting of sebacic acid, succinic acid, and combinations thereof.

6. 3. The polymeric delivery system of claim 2, wherein the at least one charged polymer and the at least one biologic are linked by a ligand interaction or complex coordination.

7. 3. The polymeric delivery system of claim 2, wherein the at least one biologic is selected from the group consisting of a nucleic acid, an amino acid, a peptide, a protein, a gene editing system, an antibody, and a cytokine.

8. The polymeric delivery system of claim 7 , wherein the at least one biologic comprises a nucleic acid.

9. The polymeric delivery system of claim 2 , wherein the at least one charged polymer is non-immunogenic.

10. 3. The polymeric delivery system of claim 2, further comprising an additive selected from the group consisting of a salt, a buffer, a surfactant, a stabilizer, and combinations thereof, wherein the additive adjusts at least one property of the polyplex.

11. 2. The polymeric delivery system of claim 1, wherein the nanoparticles are self-assembled particles and the copolymers are block copolymers, each block copolymer comprising a first block of the polyester copolymer of polyol and polycarboxylic acid and a second block comprising a second monomer or a second polymer.

12. The first block is represented by formula (1): -[-CO-(CH ２ ) ｍ -R-(CH ２ ) ｎ -CO ２ -CH ２ -CH(OR’)-CH ２ -O-] ｘ (1) wherein m and n are integers from 1 to 30, x is an integer from 1 to 75, and R is —CH ２ -, -NH-, -NCH ３ -, -O-, -S-S-, -CH=CH-, -C≡C-, -CO ２ -, and -CONH-, where R' is hydrogen or another replaceable pendant group.

12. The polymeric delivery system of claim 11, having the chemical structure:

13. 12. The polymeric delivery system of claim 11, wherein the self-assembled particles have a number-weighted average particle size ranging from about 10 nm to about 1000 nm.

14. The polymeric delivery system of claim 11 further comprising a solvent.

15. The polymeric delivery system of claim 11 , wherein the first block is substantially unbranched.

16. 12. The polymeric delivery system of claim 11, wherein at least one polyol of the first block is modified to include an alkyl tail at least 5 carbons in length.

17. The polymeric delivery system of claim 11 , wherein the self-assembled particle is a polymersome.

18. The polymeric delivery system of claim 11 , wherein the self-assembled particles are polymeric micelles.

19. The polymeric delivery system of claim 11 , wherein the self-assembled particles are polyplexes.

20. 12. The polymeric delivery system of claim 11, wherein the block copolymer and the biologic are linked by a ligand interaction or complex coordination.

21. The polymeric delivery system of claim 11 , wherein the block copolymer is cationic and the biologic is anionic.

22. 12. The polymeric delivery system of claim 11, wherein the biologic is selected from the group consisting of a nucleic acid, an amino acid, a peptide, a protein, a gene editing system, an antibody, and a cytokine.

23. 23. The polymeric delivery system of claim 22, wherein the biologic comprises a nucleic acid.

24. A method for forming a polymeric delivery system, comprising combining a polymer and a biological agent in a solvent to form nanoparticles, each nanoparticle comprising at least one polymer and at least one biological agent, and each polymer comprising a polyester copolymer of a polyol and a polycarboxylic acid.

25. 25. The method of claim 24, wherein the nanoparticles are polyplexes, the polymers are charged polymers, and the method comprises combining the charged polymers with the biologic in the solvent to electrostatically associate the charged polymer and the biologic as the polyplexes, each of the charged polymers comprising the polyester copolymer of a polyol and a polycarboxylic acid modified with at least one charged moiety having an opposite charge to the net charge of the biologic.

26. 26. The method of claim 25, wherein the polyplexes have a number weighted average particle size ranging from about 10 nm to about 1000 nm.

27. 26. The method of claim 25, wherein the polyester copolymer of a polyol and a polycarboxylic acid comprises a copolymer of glycerol and a polycarboxylic acid selected from the group consisting of sebacic acid, succinic acid, and combinations thereof.

28. 26. The method of claim 25, further comprising modifying said polyester copolymer of polyol and polycarboxylic acid with said charged moiety to form said charged polymer.

29. 26. The method of claim 25, wherein the polyplexes have a number weighted average particle size ranging from about 10 nm to about 1000 nm.

30. 26. The method of claim 25, wherein the biologic is selected from the group consisting of a nucleic acid, an amino acid, a peptide, a protein, a gene editing system, an antibody, and a cytokine.

31. 31. The method of claim 30, wherein the biologic comprises a nucleic acid.

32. 26. The method of claim 25, further comprising adding at least one additive to the polymeric delivery system, wherein the additive is selected from the group consisting of a salt, a buffer, a surfactant, a stabilizer, and combinations thereof.

33. 25. The method of claim 24, wherein the nanoparticles are self-assembled particles and the polymers are block copolymers, each block copolymer comprising a first block of a polyester copolymer of a polyol and a polycarboxylic acid and a second block of a second monomer or second polymer.

34. 34. The method of claim 33, wherein the self-assembled particles have a number weighted average particle size in the range of about 10 nm to about 1000 nm.

35. 34. The method of claim 33, wherein the self-assembled particle is a polymersome.

36. 34. The method of claim 33, wherein the self-assembled particles are polymeric micelles.

37. 34. The method of claim 33, wherein the self-assembled particles are polyplexes.

38. 34. The method of claim 33, wherein the biologic is selected from the group consisting of a nucleic acid, an amino acid, a peptide, a protein, a gene editing system, an antibody, and a cytokine.

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