Polymeric excipients containing N,N - diethylacrylamide (DEA) for biopharmaceutical formulations

Abstract

The present disclosure provides a polyacrylamide-based copolymer excipient comprising a water-soluble carrier monomer having an acrylamide-reactive moiety and a functional dopant monomer comprising N,N-diethylacrylamide (DEA). The copolymer can reduce or prevent aggregation of biomolecules and lipid-based vehicles in aqueous formulations at hydrophobic interfaces. Thus, biomolecules and lipid-based vehicles in formulations containing the polyacrylamide-based copolymer can exhibit increased stability, such as increased thermal stability, compared to the same formulations that do not contain the polyacrylamide-based copolymer. The polyacrylamide-based copolymer can be used with any biomolecule or lipid-based vehicle that tends to aggregate in an aqueous medium.

Description

Technical Field

[0001] 1. Introduction Many biopharmaceuticals in aqueous formulations are prone to irreversible aggregation when exposed to high temperatures and / or agitation and require careful storage and refrigerated transportation (cold chain) to maintain their activity over their shelf life. Maintaining the integrity of various aggregation-prone biopharmaceuticals presents challenges to the pharmaceutical industry, healthcare providers, and people in the world who need treatment with such biopharmaceuticals. The mechanisms of aggregation vary among different biopharmaceutical compounds, but many contain hydrophobic patches or are partially denatured, exposing hydrophobic segments of the molecule that drive absorption to the interface (e.g., air-water, glass-water, or rubber-water interfaces). The high local concentration of molecules at these interfaces, combined with interface-mediated partial unfolding, causes initial nucleation of aggregation events that allow further aggregation. This tendency to aggregate at the interface increases with formulation concentration and adversely affects the stability of the entire formulation.

Background Art

[0002] Excipients are often overlooked as the "inactive" components of pharmaceutical formulations. However, in many formulations, excipients play an important role in drug bioavailability by improving the solubility, absorbability, and stability of the active ingredient. Approaches to stabilizing biopharmaceuticals would use excipients that can be easily and stably incorporated into high-concentration commercial formulations without interacting with the therapeutic agent in bulk, without changing its pharmacokinetics, or without increasing its toxicity.

[0003] Therefore, there is a need for improved biopharmaceutical formulations.

Summary of the Invention

[0004] 2. Summary The present disclosure provides a polyacrylamide-based copolymer excipient comprising a water-soluble carrier monomer having an acrylamide-reactive moiety and a functional dopant monomer comprising N,N-diethylacrylamide (DEA). The copolymer can reduce or prevent aggregation of biomolecules and lipid-based vehicles in aqueous formulations at hydrophobic interfaces. Thus, biomolecules and lipid-based vehicles in formulations containing the polyacrylamide-based copolymer can exhibit increased stability, such as increased thermal stability, compared to the same formulations without the polyacrylamide-based copolymer. The polyacrylamide-based copolymer can be used with any biomolecule or lipid-based vehicle that is prone to aggregation in an aqueous medium, and these include, but are not limited to, proteins such as antibodies and fragments thereof, cytokines, chemokines, hormones, vaccine antigens, cancer antigens, adjuvants, and combinations thereof.

[0005] The polyacrylamide-based copolymer can also be used with lipid-based vehicles that are prone to aggregation in an aqueous medium to prevent or reduce aggregation, and lipid-based vehicles include, but are not limited to, liposomes, lipid nanoparticles, polymerosomes, and micelles.

[0006] According to some embodiments, the polyacrylamide-based copolymer excipient provides a stable formulation of the biocompound. In some embodiments, these excipients are synthetic copolymers composed of a water-soluble carrier monomer selected to assist polymer solubility and a functional dopant monomer containing N,N-diethylacrylamide (DEA). The DEA monomer is hypothesized to promote polymer-interface interactions, reduce the interaction of biomolecules at the interface, and thus improve the stability of biopharmaceutical formulations. In some embodiments, a library of polyacrylamide-based copolymers containing DEA is created using high-precision high-throughput synthesis using reversible addition-fragmentation chain transfer (RAFT) polymerization. In the present disclosure, the polyacrylamide-based copolymer can provide a stable formulation of a biopharmaceutical formulation that exhibits significantly faster pharmacokinetics than the corresponding commercially available biopharmaceutical formulation.

[0007] The copolymer excipients described herein can also be more widely applied to improve the thermal stability of biopharmaceutical formulations as simple "drop-in" excipients without changing bioactivity, pharmacokinetics or pharmacodynamics. For example, the copolymer excipients described herein can be used as drop-in excipients in combination with other formulation approaches intended to modify or regulate the pharmacokinetics of protein formulations.

[0008] A polyacrylamide-based copolymer comprising a water-soluble carrier monomer containing an acrylamide-reactive moiety; and a functional dopant monomer containing N,N-diethylacrylamide (DEA), wherein the weight percentage (wt%) of the water-soluble carrier monomer is from 70% to 98%, and the weight percentage (wt%) of the functional dopant monomer is from 2% to 30%, and the average molecular weight (Mn) of the polyacrylamide-based copolymer is from 1,000 g / mol to 30,000 g / mol, and the degree of polymerization is from 10 to 250, is also provided in the present disclosure.

[0009] As disclosed herein, high-throughput controlled radical polymerization techniques can be implemented to create a library of polyacrylamide-based copolymer excipients, including those containing monomers of N,N-diethylacrylamide (DEA). Non-limiting examples of polyacrylamide-based copolymers provided herein include water-soluble carrier monomers selected from N-(3-methoxypropyl)acrylamide (MPAM), 4-acryloylmorpholine (MORPH), N,N-dimethylacrylamide (DMA), N-hydroxyethylacrylamide (HEAM), and acrylamide (AM), and polyacrylamide-based copolymers containing functional dopant monomers including DEA. In some embodiments, the functional dopant monomer includes DEA and further includes a functional dopant monomer selected from N-[tris(hydroxymethyl)-methyl]acrylamide (TRI), 2-acrylamido-2-methylpropanesulfonic acid (AMP), (3-acrylamidopropyl)trimethylammonium chloride (TMA), N-isopropylacrylamide (NIP), N-tert-butylacrylamide (TBA), and N-phenylacrylamide (PHE).

[0010] Provided in the present disclosure is a composition comprising from 0.005 wt% to 0.2 wt% of a polyacrylamide-based copolymer comprising from 70 wt% to 95 wt% of a MORPH carrier monomer and from 5 wt% to 30 wt% of a DEA dopant monomer.

[0011] Also provided is a method of increasing the stability of a formulation containing a biomolecule, comprising adding from 0.005 wt% to 5 wt% of the polyacrylamide-based copolymer of the present disclosure to the formulation.

[0012] Also provided is a method of increasing the stability of a protein formulation, comprising adding from 0.005 wt% to 5 wt% of the polyacrylamide-based copolymer of the present disclosure to the protein formulation.

[0013] A method for increasing the stability of a formulation containing a lipid-based vehicle is also provided, which includes adding from about 0.005 wt% to about 5 wt% of the polyacrylamide-based copolymer of the present disclosure to the formulation.

[0014] A method for reducing the aggregation rate of biomolecules in an aqueous composition is also provided, which includes adding from 0.005 wt% to 5 wt% of the polyacrylamide-based copolymer of the present disclosure to the formulation.

[0015] A method for reducing the aggregation rate of proteins in an aqueous composition is also provided, which includes adding from 0.005 wt% to about 5 wt% of the polyacrylamide-based copolymer of the present disclosure to the protein formulation.

[0016] A method for reducing the aggregation rate of lipid-based vehicles in an aqueous composition is also provided, which includes adding from 0.005 wt% to 5 wt% of the polyacrylamide-based copolymer of the present disclosure to the formulation.

BEST MODE FOR CARRYING OUT THE INVENTION

[0017] 3. DETAILED DESCRIPTION As summarized above, the present disclosure describes a library of copolymers that include different water-soluble acrylamide monomers and a functional dopant monomer including N,N-diethylacrylamide (DEA) at various weight ratios, molecular weights, and degrees of polymerization. The copolymers of the present disclosure can reduce or prevent the aggregation of biomolecules (e.g., proteins and peptides) or lipid-based vehicles in aqueous formulations at hydrophobic interfaces, particularly molecules that tend to aggregate in aqueous media. Proteins and other biomolecules are often used in the treatment of a wide variety of diseases and disorders, but maintaining the stability of such formulations and preventing the aggregation of molecules is an important challenge faced by the biopharmaceutical industry. Proteins, other biomolecules, and lipid-based vehicles, including but not limited to liposomes, micelles, polymerosomes, and lipid nanoparticles, can aggregate due to various factors including heat stress, chemical degradation, or exposure to surfaces and interfaces.

[0018] Non-covalent physical aggregation is mediated by forces such as hydrophobic interactions, van der Waals interactions, hydrogen bonds, and electrostatic forces. The adsorption of such molecules onto various interfaces, particularly the air-water interface, plays an important role in inducing aggregation. These aggregates can elicit an immunogenic response in the body and lead to the production of anti-therapeutic antibodies.

[0019] The polyacrylamide-based copolymers of the present disclosure can reduce or prevent the aggregation of biomolecules and lipid-based vehicles in aqueous formulations, resulting in increased stability. The polyacrylamide-based copolymers can be used with any molecule that is prone to aggregation in an aqueous medium, and they include, but are not limited to, proteins such as antibodies and their fragments, cytokines, chemokines, hormones, vaccine antigens, cancer antigens, adjuvants, and combinations thereof.

[0020] The polyacrylamide-based copolymers of the present disclosure can also be used with lipid-based vehicles that are prone to aggregation in an aqueous medium to prevent or reduce aggregation, and the lipid-based vehicles include, but are not limited to, liposomes, lipid nanoparticles, polymerosomes, and micelles.

[0021] The polyacrylamide-based copolymers of the present disclosure prevent the aggregation of proteins, other biomolecules, and lipid-based vehicles such as liposomes, lipid nanoparticles, polymerosomes, and micelles by providing an inert barrier at the hydrophobic interface of the aqueous formulation to prevent intermolecular interactions such as protein-protein interactions. In some embodiments, the hydrophobic interface is the air-water interface. In some embodiments, the hydrophobic interface is an encapsulation-water interface including, but not limited to, a glass-water interface, a rubber-water interface, a plastic-water interface, or a metal-water interface. In some embodiments, the hydrophobic interface is an oil-water interface.

[0022] As described herein, the subject copolymers can act as stabilizers for formulations containing biomolecules. In some embodiments, the formulation is a protein formulation. Polyacrylamide-based copolymers can also act as stabilizers for formulations containing lipid-based vehicles that are prone to aggregation in aqueous media including, but not limited to, liposomes, lipid nanoparticles, polymerosomes and micelles. In some embodiments, copolymers including N,N-diethylacrylamide (DEA) copolymers were identified using high-throughput screening of a large library of combinatorial acrylamide-based copolymer excipients. In some embodiments, these copolymers enhance the stability of the formulation without exerting any modifying effect on the molecules of the formulation. For example, the copolymers can enhance the stability of protein formulations without a protein-modifying effect. In some embodiments, copolymers comprising water-soluble “carrier” monomers and functional “dopant” monomers act as stabilizing excipients to reduce the interaction of biomolecules with interfaces such as the air-liquid interface. In some embodiments, the biomolecule is a protein. In some embodiments, copolymers comprising water-soluble “carrier” monomers and functional “dopant” monomers act as stabilizing excipients to reduce the interaction of lipid-based vehicles with interfaces such as the air-liquid interface.

[0023] Accordingly, the present disclosure provides a method for reducing the aggregation of biomolecules or lipid-based vehicles comprising the polyacrylamide-based copolymer excipients disclosed herein. In some embodiments, the biomolecule is a protein. In some embodiments, the protein is selected from antibodies and their fragments, cytokines, chemokines, hormones, vaccine antigens, cancer antigens, adjuvants, and combinations thereof. In some embodiments, the biomolecule is a nucleic acid. In some embodiments, the lipid-based vehicle is a liposome, micelle, polymerosome or lipid nanoparticle. The polyacrylamide-based copolymer of the present disclosure comprises a water-soluble carrier monomer comprising an acrylamide-reactive moiety and a functional dopant monomer comprising an acrylamide-reactive moiety. In some embodiments, the copolymer comprises a DEA dopant monomer and an MPAM or MORPH carrier monomer.

[0024] A method for increasing the stability of a formulation containing a biomolecule or lipid-based vehicle is also provided. In some embodiments, a method for increasing the thermal stability of a formulation containing a biomolecule or lipid-based vehicle is provided. In other embodiments, a method for reducing the aggregation rate of a biomolecule or lipid-based vehicle in an aqueous composition is provided. The method comprises adding the copolymer of the present disclosure to the formulation.

[0025] 3.1. Pharmaceutical Composition As summarized above, the present disclosure provides a pharmaceutical composition comprising a polyacrylamide-based copolymer comprising a water-soluble carrier monomer and a functional dopant monomer comprising N,N-diethylacrylamide (DEA). In some embodiments, the polyacrylamide-based copolymer is amphiphilic.

[0026] 3.1.1. Polyacrylamide-based Copolymer The present disclosure provides a polyacrylamide-based copolymer. In some embodiments, the polyacrylamide-based copolymer comprises a water-soluble carrier monomer and a functional dopant monomer comprising N,N-diethylacrylamide (DEA). In some embodiments, the polyacrylamide-based copolymer is amphiphilic.

[0027] In some embodiments, the polyacrylamide-based copolymer comprises a nonionic water-soluble acrylamide monomer and a functional dopant monomer that is N,N-diethylacrylamide (DEA). In some embodiments, the polyacrylamide-based copolymer further comprises a functional acrylamide dopant monomer selected from a hydrophobic functional acrylamide dopant monomer, an aromatic functional acrylamide dopant monomer, a hydrogen-bonding functional acrylamide dopant monomer, and an ionic functional acrylamide dopant monomer.

[0028] The polyacrylamide copolymer of the present disclosure includes a water-soluble carrier monomer. In some embodiments, the water-soluble carrier monomer is nonionic. In some embodiments, the water-soluble carrier monomer is selected from N-(3-methoxypropyl)acrylamide (MPAM), 4-acryloylmorpholine (MORPH), N,N-dimethylacrylamide (DMA), N-hydroxyethylacrylamide (HEAM), and acrylamide (AM), or combinations thereof. In some embodiments, the water-soluble carrier monomer is selected from MPAM and MORPH. In some embodiments, the water-soluble carrier monomer is N-(3-methoxypropyl)acrylamide (MPAM). In some embodiments, the water-soluble carrier monomer is 4-acryloylmorpholine (MORPH). In some embodiments, the water-soluble carrier monomer is N,N-dimethylacrylamide (DMA). In some embodiments, the water-soluble carrier monomer is N-hydroxyethylacrylamide (HEAM). In some embodiments, the water-soluble carrier monomer is acrylamide (AM). In some embodiments, the copolymer includes a water-soluble carrier monomer selected from N-(3-methoxypropyl)acrylamide (MPAM) and 4-acryloylmorpholine (MORPH).

[0029] The polyacrylamide-based copolymer of the present disclosure also includes a functional dopant monomer containing N,N-diethylacrylamide (DEA), for example, a functional dopant monomer consisting essentially of DEA or consisting of DEA. In some embodiments, the polyacrylamide-based copolymer further includes an additional hydrophobic functional acrylamide dopant monomer. In some embodiments, the hydrophobic functional acrylamide dopant monomer is N-isopropylacrylamide (NIP) or N-tert-butylacrylamide (TBA). In some embodiments, the hydrophobic functional acrylamide dopant monomer is N-isopropylacrylamide (NIP). In some embodiments, the hydrophobic functional acrylamide dopant monomer is N-tert-butylacrylamide (TBA). In some embodiments, the polyacrylamide-based copolymer further includes an aromatic functional acrylamide dopant monomer. In some embodiments, the aromatic functional acrylamide dopant monomer is N-phenylacrylamide (PHE). In some embodiments, the polyacrylamide-based copolymer further includes a hydrogen-bonding functional acrylamide dopant monomer. In some embodiments, the hydrogen-bonding functional acrylamide dopant monomer is N-[tris(hydroxymethyl)methyl]acrylamide (TRI). In some embodiments, the polyacrylamide-based copolymer further includes an ionic functional acrylamide dopant monomer. In some embodiments, the ionic functional acrylamide dopant monomer is 2-acrylamido-2-methylpropanesulfonic acid (AMP) or (3-acrylamidopropyl)trimethylammonium chloride (TMA). In some embodiments, the ionic functional acrylamide dopant monomer is 2-acrylamido-2-methylpropanesulfonic acid (AMP). In some embodiments, the ionic functional acrylamide dopant monomer is (3-acrylamidopropyl)trimethylammonium chloride (TMA).

[0030] In some embodiments, the polyacrylamide-based copolymer comprises a water-soluble carrier monomer selected from N-(3-methoxypropyl)acrylamide (MPAM), 4-acryloylmorpholine (MORPH), N,N-dimethylacrylamide (DMA), N-hydroxyethylacrylamide (HEAM), and acrylamide (AM), and a functional dopant monomer comprising N,N-diethylacrylamide (DEA).

[0031] In some embodiments, the water-soluble carrier monomer is N-(3-methoxypropyl)acrylamide (MPAM). In some embodiments, the water-soluble carrier monomer is N-(3-methoxypropyl)acrylamide (MPAM) and the functional dopant monomer is N,N-diethylacrylamide (DEA).

[0032] In some embodiments, the water-soluble carrier monomer is 4-acryloylmorpholine (MORPH). In some embodiments, the water-soluble carrier monomer is 4-acryloylmorpholine (MORPH) and the functional dopant monomer is N,N-diethylacrylamide (DEA).

[0033] In some embodiments, the water-soluble carrier monomer is N,N-dimethylacrylamide (DMA). In some embodiments, the water-soluble carrier monomer is N,N-dimethylacrylamide (DMA) and the functional dopant monomer is N,N-diethylacrylamide (DEA).

[0034] In some embodiments, the water-soluble carrier monomer is N-hydroxyethylacrylamide (HEAM). In some embodiments, the water-soluble carrier monomer is N-hydroxyethylacrylamide (HEAM) and the functional dopant monomer is N,N-diethylacrylamide (DEA).

[0035] In some embodiments, the water-soluble carrier monomer is acrylamide (AM). In some embodiments, the water-soluble carrier monomer is acrylamide (AM), and the functional dopant monomer is N,N-diethylacrylamide (DEA).

[0036] In some embodiments, the polyacrylamide-based copolymer comprises N-(3-methoxypropyl)acrylamide (MPAM) or 4-acryloylmorpholine (MORPH) as the water-soluble carrier monomer and N,N-diethylacrylamide (DEA) as the functional dopant monomer.

[0037] In some embodiments, the polyacrylamide-based copolymer comprises N,N-dimethylacrylamide (DMA), N-hydroxyethylacrylamide (HEAM) or acrylamide (AM) as the water-soluble carrier monomer, and the functional dopant monomer is N,N-diethylacrylamide (DEA).

[0038] In some embodiments, the amount of the functional dopant monomer used in the copolymerization reaction is designed to maximize the dopant loading while producing a functional copolymer having a lower critical solution temperature (LCST) value above 37°C. In some embodiments, this results in a copolymer that remains soluble at all relevant temperatures. In some embodiments, the polyacrylamide-based copolymer comprises from about 2 wt% to about 30 wt% of the functional dopant monomer, such as from about 5 wt% to about 30 wt%, from about 10 wt% to about 30 wt%, from about 15 wt% to about 30 wt%, from about 20 wt% to about 30 wt%, from about 25 wt% to about 30 wt%, from about 2 wt% to about 25 wt%, from about 5 wt% to about 25 wt%, from about 10 wt% to about 25 wt%, from about 15 wt% to about 25 wt%, from about 20 wt% to about 25 wt%, from about 2 wt% to about 20 wt%, from about 5 wt% to about 20 wt%, from about 10 wt% to about 20 wt%, from about 15 wt% to about 20 wt%, from about 2 wt% to about 15 wt%, from about 5 wt% to about 15 wt%, from about 10 wt% to about 15 wt%, from about 2 wt% to about 10 wt%, from about 5 wt% to about 10 wt%, or from about 2 wt% to about 5 wt% of the functional dopant monomer. In some embodiments, the polyacrylamide-based copolymer comprises about 2 wt%, about 5 wt%, about 8 wt%, about 10 wt%, about 12 wt%, about 15 wt%, about 18 wt%, about 20 wt%, about 22 wt%, about 25 wt%, about 28 wt%, or about 30 wt% of the functional dopant monomer. In some embodiments, the polyacrylamide-based copolymer comprises from 2 wt% to 30 wt% of the functional dopant monomer, such as from 5 wt% to 30 wt%, from 10 wt% to 30 wt%, from 15 wt% to 30 wt%, from 20 wt% to 30 wt%, from 25 wt% to 30 wt%, from 2 wt% to 25 wt%, from 5 wt% to 25 wt%, from 10 wt% to 25 wt%, from 15 wt% to 25 wt%, from 20 wt% to 25 wt%, from 2 wt% to 20 wt%, from 5 wt% to 20 wt%, from 10 wt% to 20 wt%, from 15 wt% to 20 wt%, from 2 wt% to 15 wt%, from 5 wt% to 15 wt%, from 10 wt% to 15 wt%, from 2 wt% to 10 wt%, from 5 wt% to 10 wt%, or from 2 wt% to 5 wt% of the functional dopant monomer.In some embodiments, the polyacrylamide-based copolymer comprises 2 wt%, 5 wt%, 8 wt%, 10 wt%, 12 wt%, 15 wt%, 18 wt%, 20 wt%, 22 wt%, 25 wt%, 28 wt%, or 30 wt% of a functional dopant monomer. In some embodiments, the polyacrylamide-based copolymer comprises 2 wt%, 5 wt%, 8 wt%, 10 wt%, 12 wt%, 15 wt%, 18 wt%, 20 wt%, 22 wt%, 25 wt%, 28 wt%, or 30 wt% of N,N-diethylacrylamide (DEA).

[0039] In some embodiments, the polyacrylamide copolymer comprises from about 70 wt% to about 98 wt% of a water-soluble carrier monomer, such as from about 75 wt% to about 98 wt%, from about 80 wt% to about 98 wt%, from about 85 wt% to about 98 wt%, from about 90 wt% to about 98 wt%, from about 95 wt% to about 98 wt%, from about 70 wt% to about 95 wt%, from about 75 wt% to about 95 wt%, from about 80 wt% to about 95 wt%, from about 85 wt% to about 95 wt%, from about 90 wt% to about 95 wt%, from about 70 wt% to about 90 wt%, from about 75 wt% to about 90 wt%, from about 80 wt% to about 90 wt%, from about 85 wt% to about 90 wt%, from about 70 wt% to about 85 wt%, from about 75 wt% to about 85 wt%, from about 80 wt% to about 85 wt%, from about 70 wt% to about 80 wt%, from about 75 wt% to about 80 wt%, or from about 70 wt% to about 75 wt% of a water-soluble carrier monomer. In some embodiments, the polyacrylamide copolymer comprises about 70 wt%, about 72 wt%, about 75 wt%, about 78 wt%, about 80 wt%, about 82 wt%, about 85 wt%, about 88 wt%, about 90 wt%, about 92 wt%, about 95 wt%, or about 98 wt% of a water-soluble carrier monomer. In some embodiments, the polyacrylamide copolymer comprises from 70 wt% to 98 wt% of a water-soluble carrier monomer, such as from 75 wt% to 98 wt%, from 80 wt% to 98 wt%, from 85 wt% to 98 wt%, from 90 wt% to 98 wt%, from 95 wt% to 98 wt%, from 70 wt% to 95 wt%, from 75 wt% to 95 wt%, from 80 wt% to 95 wt%, from 85 wt% to 95 wt%, from 90 wt% to 95 wt%, from 70 wt% to 90 wt%, from 75 wt% to 90 wt%, from 80 wt% to 90 wt%, from 85 wt% to 90 wt%, from 70 wt% to 85 wt%, from 75 wt% to 85 wt%, from 80 wt% to 85 wt%, from 70 wt% to 80 wt%, from 75 wt% to 80 wt%, or from 70 wt% to 75 wt% of a water-soluble carrier monomer. In some embodiments, the polyacrylamide copolymer comprises 70 wt%, 72 wt%, 75 wt%, 78 wt%, 80 wt%, 82 wt%, 85 wt%, 88 wt%, 90 wt%, 92 wt%, 95 wt%, or 98 wt% of a water-soluble carrier monomer.

[0040] In some embodiments, the polyacrylamide copolymer comprises from about 70 wt% to about 98 wt% of a water-soluble carrier monomer and from about 2 wt% to about 30 wt% of a functional dopant monomer. For example, the polyacrylamide copolymer can comprise from about 70 wt% to about 98 wt%, from about 70 wt% to about 95 wt%, from about 70 wt% to about 80 wt%, from about 80 wt% to about 90 wt%, from about 90 wt% to about 98 wt%, about 70 wt%, about 75 wt%, about 80 wt%, about 85 wt%, about 90 wt%, about 95 wt%, or about 98 wt% of a water-soluble carrier monomer and from about 2 wt% to about 30 wt%, from about 5 wt% to about 25 wt%, from about 5 wt% to about 20 wt%, from about 2 wt% to about 5 wt%, from about 5 wt% to about 10 wt%, from about 10 wt% to about 15 wt%, from about 15 wt% to about 20 wt%, from about 20 wt% to about 25 wt%, or from about 25 wt% to about 30 wt%, about 2 wt%, about 5 wt%, about 8 wt%, about 10 wt%, about 12 wt%, about 15 wt%, about 18 wt%, about 20 wt%, about 22 wt%, about 25 wt%, about 28 wt%, or about 30 wt% of a functional dopant monomer.

[0041] In some embodiments, the polyacrylamide copolymer comprises from 70 wt% to 98 wt% of a water-soluble carrier monomer and from 2 wt% to 30 wt% of a functional dopant monomer. For example, the polyacrylamide copolymer can comprise from 70 wt% to 98 wt%, from 70 wt% to 95 wt%, from 70 wt% to 80 wt%, from 80 wt% to 90 wt%, from 90 wt% to 98 wt%, 70 wt%, 75 wt%, 80 wt%, 85 wt%, 90 wt%, 95 wt% or 98 wt% of a water-soluble carrier monomer and from 2 wt% to 30 wt%, from 5 wt% to 25 wt%, from 5 wt% to 20 wt%, from 2 wt% to 5 wt%, from 5 wt% to 10 wt%, from 10 wt% to 15 wt%, from 15 wt% to 20 wt%, from 20 wt% to 25 wt% or from 25 wt% to 30 wt%, 2 wt%, 5 wt%, 8 wt%, 10 wt%, 12 wt%, 15 wt%, 18 wt%, 20 wt%, 22 wt%, 25 wt%, 28 wt% or 30 wt% of a functional dopant monomer.

[0042] In some embodiments, the polyacrylamide copolymer comprises from about 2 wt% to about 30 wt% of the functional dopant monomer DEA. In some embodiments, the polyacrylamide copolymer comprises from about 5 wt% to about 30 wt% of the functional dopant monomer DEA. In some embodiments, the polyacrylamide copolymer comprises from about 10 wt% to about 28 wt% of the functional dopant monomer DEA. In some embodiments, the polyacrylamide copolymer comprises from about 5 wt% to about 26 wt% of the functional dopant monomer DEA. In some embodiments, the polyacrylamide copolymer comprises from about 5 wt% to about 10 wt% of the functional dopant monomer DEA. In some embodiments, the polyacrylamide copolymer comprises from about 10 wt% to about 15 wt% of the functional dopant monomer DEA. In some embodiments, the polyacrylamide copolymer comprises from about 15 wt% to about 20 wt% of the functional dopant monomer DEA. In some embodiments, the polyacrylamide copolymer comprises from about 20 wt% to about 26 wt% of the functional dopant monomer DEA.

[0043] In some embodiments, the polyacrylamide copolymer comprises from 2 wt% to 30 wt% of the functional dopant monomer DEA, such as from 5 wt% to 30 wt%, from 10 wt% to 28 wt%, from 5 wt% to 26 wt%, from 5 wt% to 10 wt%, from 10 wt% to 15 wt%, from 15 wt% to 20 wt%, or from 20 wt% to 26 wt% of the functional dopant monomer DEA.

[0044] In some embodiments, the polyacrylamide-based copolymer comprises MORPH as a water-soluble carrier monomer and from about 2 wt% to about 30 wt% DEA as a functional dopant monomer. In some embodiments, the copolymer comprises MORPH and from about 5 wt% to about 30 wt% DEA. In some embodiments, the copolymer comprises MORPH and from about 10 wt% to about 28 wt% DEA. In some embodiments, the copolymer comprises MORPH and from about 5 wt% to about 26 wt% DEA. In some embodiments, the copolymer comprises MORPH and from about 5 wt% to about 10 wt% DEA. In some embodiments, the copolymer comprises MORPH and from about 10 wt% to about 15 wt% DEA. In some embodiments, the copolymer comprises MORPH and from about 15 wt% to about 20 wt% DEA. In some embodiments, the copolymer comprises MORPH and from about 20 wt% to about 25 wt% DEA. In some embodiments, the copolymer comprises MORPH and from about 25 wt% to about 30 wt% DEA. In some embodiments, the copolymer comprises MORPH and from about 20 wt% to about 28 wt% DEA. In some embodiments, the copolymer comprises MORPH and about 21 wt% DEA. In some embodiments, the copolymer comprises MORPH and about 22 wt% DEA. In some embodiments, the copolymer comprises MORPH and about 23 wt% DEA. In some embodiments, the copolymer comprises MORPH and about 24 wt% DEA. In some embodiments, the copolymer comprises MORPH and about 25 wt% DEA.

[0045] In some embodiments, the polyacrylamide copolymer comprises MORPH as a water-soluble carrier monomer and 2 wt% to 30 wt% DEA, such as 5 wt% to 30 wt%, 10 wt% to 28 wt%, 5 wt% to 26 wt%, 5 wt% to 10 wt%, 10 wt% to 15 wt%, 15 wt% to 20 wt%, 20 wt% to 25 wt%, 25 wt% to 30 wt%, or 20 wt% to 28 wt% DEA as a functional dopant monomer. In some embodiments, the copolymer comprises MORPH and 21 wt% DEA, such as 22, 23 wt%, 24 wt% or 25 wt% DEA.

[0046] In some embodiments, the polyacrylamide copolymer comprises MPAM as a water-soluble carrier monomer and from about 2 wt% to about 16 wt% DEA as a functional dopant monomer. In some embodiments, the polyacrylamide copolymer comprises MPAM as a water-soluble carrier monomer and from about 5 wt% to about 15 wt% DEA as a functional dopant monomer. In some embodiments, the polyacrylamide copolymer comprises MPAM as a water-soluble carrier monomer and from about 6 wt% to about 10 wt% DEA as a functional dopant monomer. In some embodiments, the polyacrylamide copolymer comprises MPAM as a water-soluble carrier monomer and about 7 wt% DEA as a functional dopant monomer. In some embodiments, the polyacrylamide copolymer comprises MPAM as a water-soluble carrier monomer and about 8 wt% DEA as a functional dopant monomer. In some embodiments, the polyacrylamide copolymer comprises MPAM as a water-soluble carrier monomer and about 9 wt% DEA as a functional dopant monomer.

[0047] In some embodiments, the polyacrylamide copolymer comprises MPAM as a water-soluble carrier monomer and 2 wt% to 16 wt% of DEA, such as 5 wt% to 15 wt%, or 6 wt% to 10 wt% of DEA as a functional dopant monomer. In some embodiments, the polyacrylamide copolymer comprises MPAM as a water-soluble carrier monomer and 7 wt% of DEA as a functional dopant monomer, such as 8 wt% or 9 wt% of DEA as a functional dopant monomer.

[0048] In some embodiments, the polyacrylamide copolymer further comprises NIP, TRI, AMP, TMA, TBA, or PHE as a functional dopant monomer. In some embodiments, the NIP, AMP, TMA, TBA, or PHE functional dopant monomer is present at about 2 wt% to about 16 wt% of the copolymer. In some embodiments, the NIP, TRI, AMP, TMA, TBA, or PHE functional dopant monomer is present at about 5 wt% to about 15 wt% of the copolymer. In some embodiments, the NIP, TRI, AMP, TMA, TBA, or PHE functional dopant monomer is present at about 6 wt% to about 14 wt% of the copolymer. In some embodiments, the NIP, TRI, AMP, TMA, TBA, or PHE functional dopant monomer is present at about 12 wt% to about 15 wt% of the copolymer. In some embodiments, the NIP, TRI, AMP, TMA, TBA, or PHE functional dopant monomer is present at about 2 wt% to about 5 wt% of the copolymer. In some embodiments, the NIP, TRI, AMP, TMA, TBA, or PHE functional dopant monomer is present at about 5 wt% to about 10 wt% of the copolymer.

[0049] In some embodiments, the polyacrylamide-based copolymer further comprises NIP, AMP, TMA, TBA, or PHE as a functional dopant monomer in an amount of 2 wt% to 16 wt% of the copolymer, such as 5 wt% to 15 wt%, 6 wt% to 14 wt%, or 12 wt% to 15 wt%. In some embodiments, the NIP, TRI, AMP, TMA, TBA, or PHE functional dopant monomer is present at 2 wt% to 5 wt% of the copolymer. In some embodiments, the NIP, TRI, AMP, TMA, TBA, or PHE functional dopant monomer is present at 5 wt% to 10 wt% of the copolymer.

[0050] In some embodiments, the degree of polymerization (DP) of the polyacrylamide-based copolymer is from about 10 to about 500, from about 20 to about 200, from about 50 to about 100, from about 100 to about 200, from about 200 to about 300, from about 300 to about 400, or from about 400 to about 500, or about 50, about 70, about 100, about 120, about 150, about 170, about 200, about 220, about 250, about 270, about 300, about 320, about 350, about 370, about 400, about 420, about 450, about 470, or about 500. In some embodiments, the DP of the copolymer is about 40. In some embodiments, the DP of the copolymer is about 50. In some embodiments, the DP of the copolymer is about 60. In some embodiments, the DP of the copolymer is about 70. In some embodiments, the DP of the copolymer is about 80. In some embodiments, the DP of the copolymer is about 90. In some embodiments, the DP of the copolymer is about 100.

[0051] In some embodiments, the degree of polymerization (DP) of the polyacrylamide-based copolymer is from 10 to 500, from 20 to 200, from 50 to 100, from 100 to 200, from 200 to 300, from 300 to 400, or from 400 to 500, or 50, 70, 100, 120, 150, 170, 200, 220, 250, 270, 300, 320, 350, 370, 400, 420, 450, 470, or 500. In some embodiments, the DP of the copolymer is 40. In some embodiments, the DP of the copolymer is 50. In some embodiments, the DP of the copolymer is 60. In some embodiments, the DP of the copolymer is 70. In some embodiments, the DP of the copolymer is 80. In some embodiments, the DP of the copolymer is 90. In some embodiments, the DP of the copolymer is 100.

[0052] In some embodiments, the molecular weight of the polyacrylamide-based copolymer is from about 1,000 g / mol to about 40,000 g / mol, such as from about 1,000 g / mol to about 35,000 g / mol, from about 1,000 g / mol to about 30,000 g / mol, from about 1,000 g / mol to about 25,000 g / mol, from about 1,000 g / mol to about 20,000 g / mol, from about 1,000 g / mol to about 15,000 g / mol, from about 1,000 g / mol to about 10,000 g / mol, from about 1,000 g / mol to about 7,000 g / mol, from about 1,000 g / mol to about 6,000 g / mol, from about 1,000 g / mol to about 5,000 g / mol, from about 1,000 g / mol to about 4,000 g / mol, from about 1,000 g / mol to about 3,000 g / mol, from about 3,000 g / mol to about 40,000 g / mol, from about 3,000 g / mol to about 35,000 g / mol, from about 3,000 g / mol to about 30,000 g / mol, from about 3,000 g / mol to about 25,000 g / mol, from about 3,000 g / mol to about 20,000 g / mol, from about 3,000 g / mol to about 15,000 g / mol, from about 3,000 g / mol to about 10,000 g / mol, from about 3,000 g / mol to about 7,000 g / mol, from about 3,000 g / mol to about 6,000 g / mol, from about 3,000 g / mol to about 5,000 g / mol, from about 3,000 g / mol to about 4,000 g / mol, from about 4,000 g / mol to about 40,000 g / mol, from about 4,000 g / mol to about 35,000 g / mol, from about 4,000 g / mol to about 30,000 g / mol, from about 4,000 g / mol to about 25,000 g / mol, from about 4,000 g / mol to about 20,000 g / mol, from about 4,000 g / mol to about 15,000 g / mol, from about 4,000 g / mol to about 10,000 g / mol, from about 4,000 g / mol to about 7,000 g / mol, from about 4,000 g / mol to about 6,000 g / mol, from about 4,000 g / mol to about 5,000 g / mol, from about 5,000 g / mol to about 40,000 g / mol, from about 5,000 g / mol to about 35,000 g / mol, from about 5,000 g / mol to about 30,000 g / mol, from about 5,000 g / mol to about 25,000 g / mol, from about 5,From about 20,000 g / mol to 000 g / mol, from about 15,000 g / mol to about 5,000 g / mol, from about 10,000 g / mol to about 5,000 g / mol, from about 7,000 g / mol to about 5,000 g / mol, from about 6,000 g / mol to about 5,000 g / mol, from about 40,000 g / mol to about 6,000 g / mol, from about 35,000 g / mol to about 6,000 g / mol, from about 30,000 g / mol to about 6,000 g / mol, from about 25,000 g / mol to about 6,000 g / mol, from about 20,000 g / mol to about 6,000 g / mol, from about 15,000 g / mol to about 6,000 g / mol, from about 10,000 g / mol to about 6,000 g / mol, from about 7,000 g / mol to about 6,000 g / mol, from about 40,000 g / mol to about 7,000 g / mol, from about 35,000 g / mol to about 7,000 g / mol, from about 30,000 g / mol to about 7,000 g / mol, from about 25,000 g / mol to about 7,000 g / mol, from about 20,000 g / mol to about 7,000 g / mol, from about 15,000 g / mol to about 7,000 g / mol, from about 10,000 g / mol to about 7,000 g / mol, from about 40,000 g / mol to about 10,000 g / mol, from about 35,000 g / mol to about 10,000 g / mol, from about 30,000 g / mol to about 10,000 g / mol, from about 25,000 g / mol to about 10,000 g / mol, from about 20,000 g / mol to about 10,000 g / mol, from about 15,000 g / mol to about 10,000 g / mol, from about 40,000 g / mol to about 15,000 g / mol, from about 35,000 g / mol to about 15,000 g / mol, from about 30,000 g / mol to about 15,000 g / mol, from about 25,000 g / mol to about 15,000 g / mol, from about 20,000 g / mol to about 15,000 g / mol, from about 40,000 g / mol to about 20,000 g / mol, from about 35,000 g / mol to about 20,000 g / mol, from about 30,000 g / mol to about 20,000 g / mol, from about 25,000 g / mol to about 20,000 g / mol, from about 40,000 g / mol to about 25,000 g / mol, from about 35,000 g / mol to about 25,000 g / mol, from about 30, It should be noted that the original text seems to be incomplete or have some formatting issues as the "from about 25,000 g / mol to about 30," at the end is not a complete range expression. This translation is done based on the existing content.000 g / mol, from about 30,000 g / mol to about 40,000 g / mol, from about 30,000 g / mol to about 35,000 g / mol, or from about 35,000 g / mol to about 40,000 g / mol. In some embodiments, the molecular weight of the copolymer is from about 1,000 to about 30,000 g / mol. In some embodiments, the molecular weight of the copolymer is from about 10,000 to about 20,000 g / mol. In some embodiments, the molecular weight of the copolymer is from about 15,000 to about 20,000 g / mol. In some embodiments, the molecular weight of the copolymer is from about 20,000 to about 25,000 g / mol. The molecular weight of the copolymer is from about 25,000 to about 30,000 g / mol. In some embodiments, the molecular weight of the copolymer is from about 30,000 to about 40,000 g / mol. In some embodiments, the molecular weight of the copolymer is from about 2,000 to about 10,000 g / mol. In some embodiments, the molecular weight of the copolymer is from about 3,000 to about 7,000 g / mol. In some embodiments, the molecular weight of the copolymer is from about 4,000 to about 6,000 g / mol.,

[0053] In some embodiments, the molecular weight of the polyacrylamide-based copolymer is from 1,000 g / mol to 40,000 g / mol, such as from 1,000 g / mol to 35,000 g / mol, from 1,000 g / mol to 30,000 g / mol, from 1,000 g / mol to 25,000 g / mol, from 1,000 g / mol to 20,000 g / mol, from 1,000 g / mol to 15,000 g / mol, from 1,000 g / mol to 10,000 g / mol, from 1,000 g / mol to 7,000 g / mol, from 1,000 g / mol to 6,000 g / mol, from 1,000 g / mol to 5,000 g / mol, from 1,000 g / mol to 4,000 g / mol, from 1,000 g / mol to 3,000 g / mol, from 3,000 g / mol to 40,000 g / mol, from 3,000 g / mol to 35,000 g / mol, from 3,000 g / mol to 30,000 g / mol, from 3,000 g / mol to 25,000 g / mol, from 3,000 g / mol to 20,000 g / mol, from 3,000 g / mol to 15,000 g / mol, from 3,000 g / mol to 10,000 g / mol, from 3,000 g / mol to 7,000 g / mol, from 3,000 g / mol to 6,000 g / mol, from 3,000 g / mol to 5,000 g / mol, from 3,000 g / mol to 4,000 g / mol, from 4,000 g / mol to 40,000 g / mol, from 4,000 g / mol to 35,000 g / mol, from 4,000 g / mol to 30,000 g / mol, from 4,000 g / mol to 25,000 g / mol, from 4,000 g / mol to 20,000 g / mol, from 4,000 g / mol to 15,000 g / mol, from 4,000 g / mol to 10,000 g / mol, from 4,000 g / mol to 7,000 g / mol, from 4,000 g / mol to 6,000 g / mol, from 4,000 g / mol to 5,000 g / mol, from 5,000 g / mol to 40,000 g / mol, from 5,000 g / mol to 35,000 g / mol, from 5,000 g / mol to 30,000 g / mol, from 5,000 g / mol to 25,000 g / mol, from 5,000 g / mol to 20,000 g / mol, from 5,000 g / mol to 15,000 g / mol, from 5,000 g / mol to 10,000 g / mol, from 5,000 g / mol to 7,000 g / mol, from 5,000 g / mol to 6,000 g / mol, from 6,000 g / mol to 40,000 g / mol, from 6,000 g / mol to 35,000 g / mol, from 6,000 g / mol to 30,000 g / mol, from 6,000 g / mol to 25,000 g / mol, from 6,000 g / mol to 20,000 g / mol, from 6,000 g / mol to 15,000 g / mol, from 6,000 g / mol to 10,000 g / mol, from 6,000 g / mol to 7,000 g / mol, from 7,000 g / mol to 40,000 g / mol, from 7,000 g / mol to 35,000 g / mol, from 7,000 g / mol to 30,000 g / mol, from 7,000 g / mol to 25,000 g / mol, from 7,000 g / mol to 20,000 g / mol, from 7,000 g / mol to 15,000 g / mol, from 7,000 g / mol to 10,000 g / mol, from 10,000 g / mol to 40,000 g / mol, from 10,000 g / mol to 35,000 g / mol, from 10,000 g / mol to 30,000 g / mol, from 10,000 g / mol to 25,000 g / mol, from 10,000 g / mol to 20,000 g / mol, from 10,000 g / mol to 15,000 g / mol, from 15,000 g / mol to 40,000 g / mol, from 15,000 g / mol to 35,000 g / mol, from 15,000 g / mol to 30,000 g / mol, from 15,000 g / mol to 25,000 g / mol, from 15,000 g / mol to 20,000 g / mol, from 20,000 g / mol to 40,000 g / mol, from 20,000 g / mol to 35,000 g / mol, from 20,000 g / mol to 30,000 g / mol, from 20,000 g / mol to 25,000 g / mol, from 25,000 g / mol to 40,000 g / mol, from 25,000 g / mol to 35,000 g / mol, from 25,000 g / mol to 30,000 g / mol, from 30,000 g / mol to 40,000 g / mol, from 30,000 g / mol to 35,000 g / mol, or from 35,000 g / mol to 40,000 g / mol. In some embodiments, the molecular weight of the copolymer is from 1,000 to 30,000 g / mol. In some embodiments, the molecular weight of the copolymer is 10,It ranges from 000 to 20,000 g / mol. In some embodiments, the molecular weight of the copolymer is from 15,000 to 20,000 g / mol. In some embodiments, the molecular weight of the copolymer is from 20,000 to 25,000 g / mol. The molecular weight of the copolymer is from 25,000 to 30,000 g / mol. In some embodiments, the molecular weight of the copolymer is from 30,000 to 40,000 g / mol. In some embodiments, the molecular weight of the copolymer is from 2,000 to 10,000 g / mol. In some embodiments, the molecular weight of the copolymer is from 3,000 to 7,000 g / mol. In some embodiments, the molecular weight of the copolymer is from 4,000 to 6,000 g / mol.,

[0054] Also provided by the present disclosure is a polyacrylamide-based copolymer comprising a water-soluble carrier monomer containing an acrylamide-reactive moiety and a functional dopant monomer containing DEA. In some embodiments, the polyacrylamide-based copolymer comprises from about 70% to about 98% of a water-soluble carrier monomer having an acrylamide-reactive moiety and from about 2% to about 30% of DEA. In some embodiments, the number average molecular weight (Mn) of the copolymer is from about 1,000 g / mol to about 30,000 g / mol. In some embodiments, the degree of polymerization is from about 10 to about 250. In some embodiments, the polyacrylamide-based copolymer comprises from 70% to 98% of a water-soluble carrier monomer having an acrylamide-reactive moiety and from 2% to 30% of DEA. In some embodiments, the number average molecular weight (Mn) of the copolymer is from 1,000 g / mol to 30,000 g / mol. In some embodiments, the degree of polymerization is from 10 to 250. In some embodiments, the water-soluble carrier monomer is nonionic. In some embodiments, the copolymer is amphiphilic.

[0055] In some embodiments, the polyacrylamide copolymer comprises from about 70 wt% to about 95 wt% of the water-soluble carrier monomer MORPH and from about 5 wt% to about 30 wt% of the functional dopant monomer DEA, the number average molecular weight (Mn) of the copolymer is from about 1,000 g / mol to about 10,000 g / mol, and the degree of polymerization is from about 10 to about 100. In some embodiments, the polyacrylamide copolymer comprises from about 74 wt% to about 80 wt% of the water-soluble carrier monomer MORPH and from about 20 wt% to about 26 wt% of the functional dopant monomer DEA, the number average molecular weight (Mn) of the copolymer is from about 1,000 g / mol to about 5,000 g / mol, and the degree of polymerization is from about 10 to about 50. In some embodiments, the polyacrylamide copolymer comprises about 77 wt% of the water-soluble carrier monomer MORPH and about 23 wt% of the functional dopant monomer DEA, the number average molecular weight (Mn) of the copolymer is about 3,200 g / mol, and the degree of polymerization is about 26.

[0056] In some embodiments, the polyacrylamide copolymer comprises from 70 wt% to 95 wt% of the water-soluble carrier monomer MORPH and from 5 wt% to 30 wt% of the functional dopant monomer DEA, the number average molecular weight (Mn) of the copolymer is from 1,000 g / mol to 10,000 g / mol, and the degree of polymerization is from 10 to 100. In some embodiments, the polyacrylamide copolymer comprises from 74 wt% to 80 wt% of the water-soluble carrier monomer MORPH and from 20 wt% to 26 wt% of the functional dopant monomer DEA, the number average molecular weight (Mn) of the copolymer is from 1,000 g / mol to 5,000 g / mol, and the degree of polymerization is from 10 to 50. In some embodiments, the polyacrylamide copolymer comprises 77 wt% of the water-soluble carrier monomer MORPH and 23 wt% of the functional dopant monomer DEA, the number average molecular weight (Mn) of the copolymer is 3,200 g / mol, and the degree of polymerization is 26.

[0057] In some embodiments, the composition comprises from about 0.001 wt% to about 1 wt% of a polyacrylamide-based copolymer, such as from about 0.001 wt% to about 0.5 wt%, from about 0.001 wt% to about 0.1 wt%, from about 0.001 wt% to about 0.05 wt%, from about 0.001 wt% to about 0.03 wt%, from about 0.005 wt% to about 1 wt%, from about 0.005 wt% to about 0.5 wt%, from about 0.005 wt% to about 0.1 wt%, from about 0.005 wt% to about 0.05 wt%, from about 0.005 wt% to about 0.03 wt%, from about 0.01 wt% to about 1 wt%, from about 0.01 wt% to about 0.5 wt%, from about 0.01 wt% to about 0.1 wt%, from about 0.01 wt% to about 0.05 wt%, or from about 0.01 wt% to about 0.03 wt% of a polyacrylamide-based copolymer. In some embodiments, the composition comprises about 0.01 wt% of a polyacrylamide-based copolymer.

[0058] In some embodiments, the composition comprises from 0.001 wt% to 1 wt% of a polyacrylamide-based copolymer, such as from 0.001 wt% to 0.5 wt%, from 0.001 wt% to 0.1 wt%, from 0.001 wt% to 0.05 wt%, from 0.001 wt% to 0.03 wt%, from 0.005 wt% to 1 wt%, from 0.005 wt% to 0.5 wt%, from 0.005 wt% to 0.1 wt%, from 0.005 wt% to 0.05 wt%, from 0.005 wt% to 0.03 wt%, from 0.01 wt% to 1 wt%, from 0.01 wt% to 0.5 wt%, from 0.01 wt% to 0.1 wt%, from 0.01 wt% to 0.05 wt%, or from 0.01 wt% to 0.03 wt% of a polyacrylamide-based copolymer. In some embodiments, the composition comprises 0.01 wt% of a polyacrylamide-based copolymer.

[0059] 3.1.2. Compositions Containing Polyacrylamide-Based Copolymers Compositions containing the polyacrylamide-based copolymers described in the present disclosure are also provided. In some embodiments, the composition comprises the polyacrylamide-based copolymer of the present disclosure and a pharmaceutically acceptable excipient.

[0060] In some embodiments, the composition comprises a polyacrylamide-based copolymer of the present disclosure and a biomolecule. In some embodiments, the biomolecule is a protein. In some embodiments, the protein is a protein that is prone to aggregation in an aqueous medium. In some embodiments, the copolymer is non-charged, cationic or anionic. In some embodiments, the copolymer is amphiphilic. In some embodiments, the carrier monomer is a water-soluble species and is responsible for both maintaining solubility and providing an inert barrier to prevent aggregation of biological species, for example to prevent protein-protein interactions. In some embodiments, the water-soluble carrier monomer is the major species within the polyacrylamide-based copolymer. In some embodiments, the functional dopant monomer (e.g., DEA) is copolymerized at a lower weight percent and is statistically incorporated throughout the resulting copolymer. In some embodiments, the dopant promotes either polymer-interface interactions or polymer-protein interactions.

[0061] In some embodiments, the composition containing the polyacrylamide copolymer and the biomolecule or lipid-based vehicle of the present disclosure is from about 0.0001 wt% to about 5 wt% of the composition, such as from about 0.0001 wt% to about 4 wt%, from about 0.0001 wt% to about 3 wt%, from about 0.0001 wt% to about 2 wt%, from about 0.0001 wt% to about 1 wt%, from about 0.0001 wt% to about 0.5 wt%, from about 0.0001 wt% to about 0.4 wt%, from about 0.0001 wt% to about 0.3 wt%, from about 0.0001 wt% to about 0.2 wt%, from about 0.0001 wt% to about 0.1 wt%, from about 0.0001 wt% to about 0.05 wt%, from about 0.0001 wt% to about 0.02 wt%, from about 0.0001 wt% to about 0.01 wt%, from about 0.0001 wt% to about 0.005 wt%, from about 0.005 wt% to about 5 wt%, from about 0.005 wt% to about 4 wt%, from about 0.005 wt% to about 3 wt%, from about 0.005 wt% to about 2 wt%, from about 0.005 wt% to about 1 wt%, from about 0.005 wt% to about 0.5 wt%, from about 0.005 wt% to about 0.4 wt%, from about 0.005 wt% to about 0.3 wt%, from about 0.005 wt% to about 0.2 wt%, from about 0.005 wt% to about 0.1 wt%, from about 0.005 wt% to about 0.05 wt%, from about 0.005 wt% to about 0.02 wt%, from about 0.005 wt% to about 0.01 wt%, from about 0.01 wt% to about 5 wt%, from about 0.01 wt% to about 4 wt%, from about 0.01 wt% to about 3 wt%, from about 0.01 wt% to about 2 wt%, from about 0.01 wt% to about 1 wt%, from about 0.01 wt% to about 0.5 wt%, from about 0.01 wt% to about 0.4 wt%, from about 0.01 wt% to about 0.3 wt%, from about 0.01 wt% to about 0.2 wt%, from about 0.01 wt% to about 0.1 wt%, from about 0.01 wt% to about 0.05 wt%, from about 0.01 wt% to about 0.02 wt%, from about 0.02 wt% to about 5 wt%, from about 0.02 wt% to about 4 wt%, from about 0.02 wt% to about 3 wt%, from about 0.02 wt% to about 2 wt%, from about 0.02 wt% to about 1 wt%, from about 0.02 wt% to about 0.5 wt%, from about 0.02 wt% to about 0.4 wt%, from about 0.02 wt% to about 0.3 wt%, from about 0.02 wt% to about 0.2 wt%, from about 0.02 wt% to about 0.1 wt%, from about 0.02 wt% to about 0.05 wt%, from about 0.05 wt% to about 5 wt%, from about 0.From about 5 wt% to about 4 wt%, from about 0.05 wt% to about 3 wt%, from about 0.05 wt% to about 2 wt%, from about 0.05 wt% to about 1 wt%, from about 0.05 wt% to about 0.5 wt%, from about 0.05 wt% to about 0.4 wt%, from about 0.05 wt% to about 0.3 wt%, from about 0.05 wt% to about 0.2 wt%, from about 0.05 wt% to about 0.1 wt%, from about 0.1 wt% to about 5 wt%, from about 0.1 wt% to about 4 wt%, from about 0.1 wt% to about 3 wt%, from about 0.1 wt% to about 2 wt%, from about 0.1 wt% to about 1 wt%, from about 0.1 wt% to about 0.5 wt%, from about 0.1 wt% to about 0.4 wt%, from about 0.1 wt% to about 0.3 wt%, from about 0.1 wt% to about 0.2 wt%, from about 0.2 wt% to about 5 wt%, from about 0.2 wt% to about 4 wt%, from about 0.2 wt% to about 3 wt%, from about 0.2 wt% to about 2 wt%, from about 0.12 wt% to about 1 wt%, from about 0.2 wt% to about 0.5 wt%, from about 0.2 wt% to about 0.4 wt%, from about 0.2 wt% to about 0.3 wt%, from about 0.3 wt% to about 5 wt%, from about 0.3 wt% to about 4 wt%, from about 0.3 wt% to about 3 wt%, from about 0.3 wt% to about 2 wt%, from about 0.3 wt% to about 1 wt%, from about 0.3 wt% to about 0.5 wt%, from about 0.3 wt% to about 0.4 wt%, from about 0.4 wt% to about 5 wt%, from about 0.4 wt% to about 4 wt%, from about 0.4 wt% to about 3 wt%, from about 0.4 wt% to about 2 wt%, from about 0.4 wt% to about 1 wt%, from about 0.4 wt% to about 0.5 wt%, from about 0.5 wt% to about 5 wt%, from about 0.5 wt% to about 4 wt%, from about 0.5 wt% to about 3 wt%, from about 0.5 wt% to about 2 wt%, from about 0.5 wt% to about 1 wt%, from about 1 wt% to about 5 wt%, from about 1 wt% to about 4 wt%, from about 1 wt% to about 3 wt%, from about 1 wt% to about 2 wt%, from about 2 wt% to about 5 wt%, from about 2 wt% to about 4 wt%, from about 2 wt% to about 3 wt%, from about 3 wt% to about 5 wt%, from about 3 wt% to about 4 wt%, or from about 4 wt% to about 5 wt% copolymer concentration. In some embodiments, the copolymer concentration is from about 0.0001 wt% to about 5 wt% of the composition. In some embodiments, the copolymer concentration is from about 0.001 wt% to about 1 wt%, from about 0.005 wt% to about 0.5 wt%, from about 0.005 wt% to about 0.02 wt%, from about 0.01 wt% to about 0.2 wt%, from about 0.1 wt% to about 0.It is 4% by weight, or from about 0.2% to about 0.3% by weight. In some embodiments, the copolymer concentration is about 0.005% by weight of the composition. In some embodiments, the copolymer concentration is about 0.01% by weight of the composition. In some embodiments, the copolymer concentration is about 0.05% by weight of the composition. In some embodiments, the copolymer concentration is about 0.1% by weight of the composition. In some embodiments, the copolymer concentration is about 1% by weight of the composition.

[0062] In some embodiments, the protein is selected from antibodies and their fragments, cytokines, chemokines, hormones, vaccine antigens, cancer antigens, adjuvants, and combinations thereof. In some embodiments, the composition exhibits a reduction in protein aggregation as compared to a composition of the same protein without the copolymer. In some embodiments, the composition exhibits a reduction in protein precipitation as compared to a composition of the same protein without the copolymer. In some embodiments, the composition contains the protein at a concentration that is at least 2-fold, at least 3-fold, at least 4-fold, or at least 5-fold higher than the concentration of the same protein in a composition without the copolymer.

[0063] In some embodiments, the composition comprising the copolymer and the biomolecule of the present disclosure further comprises one or more of a pharmaceutically acceptable carrier, an aqueous buffer, an isotonicity regulator, and a preservative. As used herein, a pharmaceutically acceptable carrier, an isotonicity regulator, and a preservative are non-toxic to the recipient at the dosage and concentrations used. In some embodiments, the composition contains a buffer such as phosphate, citrate, succinate, other organic acids, and histidine, and the term "buffer" refers to a mixture of a weak acid and its conjugate base, or vice versa, and is used to maintain the pH of the solution at a substantially constant value. In some embodiments, the buffer solution contains one or more phosphates. In some embodiments, the buffer is sodium phosphate.

[0064] In some embodiments, the composition contains an isotonicity regulator such as sodium chloride, potassium chloride, mannitol, dextrose, glycerol, or magnesium chloride. In some embodiments, the isotonicity regulator is sodium chloride (NaCl) or glycerol. In some embodiments, the isotonicity regulator is glycerol.

[0065] In some embodiments, the composition contains one or more preservatives.

[0066] In some embodiments, the composition containing the copolymer of the present disclosure is an aqueous composition. In some embodiments, the composition containing the copolymer of the present disclosure essentially contains water.

[0067] In some embodiments, the pH of the composition is from about 4 to about 9, for example, from about 4 to about 8, from about 4 to about 7, from about 4 to about 6, from about 4 to about 5, from about 5 to about 9, from about 5 to about 8, from about 5 to about 7, from about 5 to about 6, from about 6 to about 9, from about 6 to about 8, from about 6 to about 7, from about 7 to about 9, from about 7 to about 8, from about 8 to about 9, or about 4, about 4.5, about 5, about 5.5, about 6, about 6.5, about 7, about 7.4, about 8, about 8.5 or about 9. In some embodiments, the pH of the composition is from about 4 to about 9. In some embodiments, the pH of the composition is from about 6 to about 8. In some embodiments, the pH of the composition is from about 7 to about 8. In some embodiments, the pH of the composition is about 7.4.

[0068] Compositions containing the polyacrylamide-based copolymer and lipid-based vehicle of the present disclosure are also provided. In some embodiments, the lipid-based vehicle is selected from liposomes, polymerosomes, micelles and lipid nanoparticles.

[0069] In some embodiments, the copolymer of the present disclosure is used as a formulation additive in a formulation containing one or more biomolecules (e.g., as described herein).

[0070] 3.1.3. Characteristics of Copolymer Compositions The polyacrylamide copolymer of the present disclosure has unique properties and can impart beneficial properties to an aqueous formulation when used in an aqueous formulation containing a biomolecule.

[0071] In some embodiments, the copolymer described in the present disclosure reduces or prevents aggregation of a biomolecule or a lipid-based vehicle, such as when formulated in an aqueous formulation containing a biomolecule or a lipid-based vehicle. In some embodiments, the biomolecule is a protein. Thus, in some embodiments, the copolymer described in the present disclosure reduces or prevents protein aggregation, such as when formulated in an aqueous formulation containing a protein.

[0072] The copolymer can be used with any protein that is prone to aggregation in an aqueous medium. Non-limiting examples include antibodies and their fragments, cytokines, chemokines, hormones, vaccine antigens, cancer antigens, adjuvants, and combinations thereof. In some embodiments, the polyacrylamide copolymer reduces or prevents aggregation of a protein that is prone to aggregation in an aqueous medium.

[0073] The copolymers of the present disclosure can also enhance the stability of formulations. According to embodiments of the present disclosure, the copolymers described herein are added to improve the stability of formulations containing, for example, biomolecules or lipid-based vehicles, such as their peptides, proteins and conjugates, nucleic acids and oligonucleotides, liposomes, polymerosomes, micelles and lipid nanoparticles. In some embodiments, the addition of the copolymers of the present disclosure increases the stability of protein formulations. In some embodiments, the addition of the copolymers of the present disclosure increases the stability of liposome formulations. In some embodiments, the addition of the copolymers of the present disclosure increases the stability of micelle formulations. In some embodiments, the addition of the copolymers of the present disclosure increases the stability of lipid nanoparticle formulations. In some embodiments, the addition of the copolymers of the present disclosure increases the stability of formulations containing one or more nucleic acids. In some embodiments, the addition of the copolymers of the present disclosure increases the stability of formulations containing one or more messenger RNAs (mRNAs). In some embodiments, the addition of the copolymers of the present disclosure increases the stability of formulations containing one or more small interfering RNAs (siRNAs). In some embodiments, the addition of the copolymers of the present disclosure increases the stability of formulations containing one or more deoxyribonucleic acids (DNAs).

[0074] In some embodiments, the copolymers of the present disclosure increase the thermal stability of formulations containing biomolecules that are, in some embodiments, proteins. Protein formulations typically require expensive refrigerated transport and storage to prevent loss of protein integrity. Maintaining protein integrity is a challenge for the pharmaceutical industry, healthcare providers, and patients worldwide, especially in developing and low-income regions where the cold chain required to maintain protein efficacy and effectiveness is incomplete, overly burdensome, or non-existent. This can result in the waste of large quantities of therapeutic protein formulations and pose a risk to patients' lives. Thus, interruption of the cold chain can be costly. The copolymers of the present disclosure can impart long-term stability and / or cold chain resilience to protein formulations. For example, commercially available protein formulations have good shelf lives when properly stored (i.e., refrigerated), but interruption of the cold chain can reduce the biological activity and integrity of the protein.

[0075] Protein formulations containing the copolymers of the present disclosure are, in some embodiments, stable for extended periods at temperatures higher than those typically required for cold chain storage. For example, protein formulations containing the copolymers of the present disclosure can be stored at room temperature (about 20°C to about 22°C) or elevated temperatures, such as about 25°C, about 30°C, about 35°C, about 40°C, about 45°C or higher, while maintaining protein integrity. In some embodiments, protein formulations containing the copolymers of the present disclosure can be stored at higher temperatures than typically required, such as about -20°C, or about 0°C, or about 2°C, or about 4°C, or about 8°C, or about 20°C, rather than at lower temperatures, such as about -80°C, or about -60°C, or about -40°C. In some embodiments, the copolymers provide protein formulations that are stored at room temperature or elevated temperatures without the need for cold chain storage. In some embodiments, cold chain storage is not required to maintain protein integrity.

[0076] In some embodiments, the use of the copolymers of the present disclosure in protein formulations maintains the integrity, bioactivity, pharmacokinetics, and pharmacodynamics of the formulation over time when exposed to conditions such as high temperature and agitation, compared to the same formulation without the copolymer. In some embodiments, the formulation maintains the integrity of the protein for about 1 day, about 1 week, about 1 month, about 3 months, about 6 months, about 9 months, about 12 months, about 18 months, about 24 months or more at a higher temperature compared to the same formulation without the copolymers of the present disclosure.

[0077] In some embodiments, the copolymers of the present disclosure reduce the aggregation rate of biomolecules in aqueous formulations. In some embodiments, the copolymers of the present disclosure reduce the aggregation rate of proteins in aqueous protein formulations. In some embodiments, the protein is a protein that tends to aggregate in an aqueous medium. In some embodiments, the protein is an antibody or fragment thereof, cytokine, chemokine, hormone, vaccine antigen, cancer antigen, adjuvant, and combinations thereof. In some embodiments, this enables the formulation of more concentrated protein solutions, thus reducing, for example, the volume of the protein solution that needs to be administered to a patient. In some embodiments, the composition contains the protein at a concentration that is at least 2-fold, at least 3-fold, at least 4-fold, or at least 5-fold higher than the concentration of the same protein in a composition without the copolymer.

[0078] Thus, the use of the copolymers of the present disclosure can impart long-term stability and / or cold-chain resilience to protein formulations. In some embodiments, the proteins of the compositions containing the copolymers of the present disclosure exhibit high stability when stored at room temperature compared to protein compositions without the copolymer. In some embodiments, the increased stability is at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 30-fold, at least 35-fold, at least 40-fold, at least 45-fold, at least 50-fold or more compared to protein compositions without the copolymer.

[0079] In some embodiments, the copolymers of the present disclosure preserve protein activity through aging under 6 months of stress without modifying one or more of the pharmacokinetics of the formulation, the secondary structure of the protein, the transparency of the formulation, and the in vivo bioactivity. In some embodiments, the copolymer reduces protein aggregation in the formulation when stored at ambient temperature (from about 23°C to about 27°C) compared to a protein formulation that does not contain the copolymer. In some embodiments, the copolymer reduces protein aggregation in the formulation when stored at 37°C compared to a protein formulation that does not contain the copolymer. In some embodiments, the copolymer reduces protein aggregation in the formulation when stored at 50°C compared to a protein formulation that does not contain the copolymer.

[0080] In some embodiments, the copolymer increases the time to aggregation of the protein formulation by at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 30-fold, at least 35-fold, at least 40-fold, at least 45-fold, or at least 50-fold when stored at 37 °C compared to a protein formulation that does not contain the copolymer. The time to aggregation can be evaluated by any known method, including, for example, a transmittance assay. In some embodiments, the copolymer increases the time to aggregation of the protein formulation by at least 10-fold when stored at 37 °C compared to a protein formulation that does not contain the copolymer. In some embodiments, the copolymer increases the time to aggregation of the protein formulation by at least 20-fold when stored at 37 °C compared to a protein formulation that does not contain the copolymer. In some embodiments, the copolymer increases the time to aggregation of the protein formulation by at least 30-fold when stored at 37 °C compared to a protein formulation that does not contain the copolymer. In some embodiments, the copolymer increases the time to aggregation of the protein formulation by at least 40-fold when stored at 37 °C compared to a protein formulation that does not contain the copolymer. In some embodiments, the copolymer increases the time to aggregation of the protein formulation by at least 50-fold when stored at 37 °C compared to a protein formulation that does not contain the copolymer. In some embodiments, the copolymer increases the time to aggregation of the protein formulation by at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 30-fold, at least 35-fold, at least 40-fold, at least 45-fold, or at least 50-fold when stored at 50 °C compared to a protein formulation that does not contain the copolymer. In some embodiments, the copolymer increases the time to aggregation of the protein formulation by at least 10-fold when stored at 50 °C compared to a protein formulation that does not contain the copolymer. In some embodiments, the copolymer increases the time to aggregation of the protein formulation by at least 20-fold when stored at 50 °C compared to a protein formulation that does not contain the copolymer.In some embodiments, the copolymer increases the time to aggregation of the protein formulation by at least 30-fold, at least 40-fold, or at least 50-fold when stored at 50°C compared to a protein formulation that does not contain the copolymer.

[0081] In addition to refrigerated transport, maintaining appropriate transport and storage conditions at the initial stage of the cold chain, during regional distribution, and at the point of patient access presents challenges in many parts of the world. As described herein, the addition of a polyacrylamide-based copolymer can maintain the integrity of protein formulations even during severe cold chain disruptions. In some embodiments, this provides a reduction in cold chain requirements for protein transport and storage that are difficult to maintain in resource-limited environments. As disclosed herein, a polyacrylamide-based copolymer as a formulation additive can improve cold chain resilience, thereby expanding global access to important drugs and vaccines. In some embodiments, the addition of the copolymer maintains the in vitro biological activity of the protein formulation for at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, or at least 6 months. In some embodiments, the addition of the copolymer maintains the in vitro biological activity of the protein formulation for at least 1 month. In some embodiments, the addition of the copolymer maintains the in vitro biological activity of the protein formulation for at least 2 months. In some embodiments, the addition of the copolymer maintains the in vitro biological activity of the protein formulation for at least 3 months.

[0082] In some embodiments, the copolymer present in the formulation containing the protein does not modify the pharmacokinetic (PK) or pharmacodynamic (PD) properties of the active ingredient in the formulation.

[0083] 3.2. Method of Use As summarized above, the present disclosure provides methods of using the compositions and co-formulations described herein that contain polyacrylamide-based copolymers containing a water-soluble carrier monomer and a functional dopant monomer (e.g., DEA).

[0084] Methods are provided for increasing the stability of formulations containing biomolecules or lipid-based vehicles. According to embodiments of the present disclosure, the copolymers described herein can be added to improve the stability of formulations containing peptides, proteins, and their conjugates, nucleic acids and oligonucleotides, liposomes, polymerosomes, micelles, and lipid nanoparticles. In some embodiments, methods are provided for increasing the stability of protein formulations. In some embodiments, methods are provided for increasing the thermal stability of formulations containing biomolecules or lipid-based vehicles. In some embodiments, methods are provided for increasing the thermal stability of protein formulations. Methods are also provided for reducing the aggregation rate of biomolecules or lipid-based vehicles in aqueous formulations. In some embodiments, the molecule is a molecule that tends to aggregate in an aqueous medium. In some embodiments, the protein is an antibody or fragment thereof, cytokine, chemokine, hormone, vaccine antigen, cancer antigen, adjuvant, and combinations thereof.

[0085] In some embodiments, the method includes adding from about 0.001 wt% to about 5 wt% of the polyacrylamide-based copolymer of the present disclosure to a formulation containing a biomolecule or a lipid-based vehicle, for example, from about 0.001 wt% to about 4 wt%, from about 0.001 wt% to about 3 wt%, from about 0.001 wt% to about 2 wt%, from about 0.001 wt% to about 1 wt%, from about 0.001 wt% to about 0.5 wt%, from about 0.001 wt% to about 0.4 wt%, from about 0.001 wt% to about 0.3 wt%, from about 0.001 wt% to about 0.2 wt%, from about 0.001 wt% to about 0.1 wt%, from about 0.001 wt% to about 0.05 wt%, from about 0.001 wt% to about 0.02 wt%, from about 0.001 wt% to about 0.01 wt%, from about 0.001 wt% to about 0.005 wt%, from about 0.005 wt% to about 5 wt%, from about 0.005 wt% to about 4 wt%, from about 0.005 wt% to about 3 wt%, from about 0.005 wt% to about 2 wt%, from about 0.005 wt% to about 1 wt%, from about 0.005 wt% to about 0.5 wt%, from about 0.005 wt% to about 0.4 wt%, from about 0.005 wt% to about 0.3 wt%, from about 0.005 wt% to about 0.2 wt%, from about 0.005 wt% to about 0.1 wt%, from about 0.005 wt% to about 0.05 wt%, from about 0.005 wt% to about 0.02 wt%, from about 0.005 wt% to about 0.01 wt%, from about 0.01 wt% to about 5 wt%, from about 0.01 wt% to about 4 wt%, from about 0.01 wt% to about 3 wt%, from about 0.01 wt% to about 2 wt%, from about 0.01 wt% to about 1 wt%, from about 0.01 wt% to about 0.5 wt%, from about 0.01 wt% to about 0.4 wt%, from about 0.01 wt% to about 0.3 wt%, from about 0.01 wt% to about 0.2 wt%, from about 0.01 wt% to about 0.1 wt%, from about 0.01 wt% to about 0.05 wt%, from about 0.01 wt% to about 0.02 wt%, from about 0.02 wt% to about 5 wt%, from about 0.02 wt% to about 4 wt%, from about 0.02 wt% to about 3 wt%, from about 0.02 wt% to about 2 wt%, from about 0.02 wt% to about 1 wt%, from about 0.02 wt% to about 0.5 wt%, from about 0.02 wt% to about 0.4 wt%, from about 0.02 wt% to about 0.3 wt%, from about 0.02 wt% to about 0.2 wt%, from about 0.02 wt% to about 0.1 wt%, from about 0.02 wt% to about 0.05 wt%, from about 0.05 wt% to about 5 wt%, from about 0.From about 4 wt% to 05 wt%, from about 3 wt% to about 0.05 wt%, from about 2 wt% to about 0.05 wt%, from about 1 wt% to about 0.05 wt%, from about 0.5 wt% to about 0.05 wt%, from about 0.4 wt% to about 0.05 wt%, from about 0.3 wt% to about 0.05 wt%, from about 0.2 wt% to about 0.05 wt%, from about 0.1 wt% to about 0.05 wt%, from about 5 wt% to about 0.1 wt%, from about 4 wt% to about 0.1 wt%, from about 3 wt% to about 0.1 wt%, from about 2 wt% to about 0.1 wt%, from about 1 wt% to about 0.1 wt%, from about 0.5 wt% to about 0.1 wt%, from about 0.4 wt% to about 0.1 wt%, from about 0.3 wt% to about 0.1 wt%, from about 0.2 wt% to about 0.1 wt%, from about 5 wt% to about 0.2 wt%, from about 4 wt% to about 0.2 wt%, from about 3 wt% to about 0.2 wt%, from about 2 wt% to about 0.2 wt%, from about 1 wt% to about 0.2 wt%, from about 0.5 wt% to about 0.2 wt%, from about 0.4 wt% to about 0.2 wt%, from about 0.3 wt% to about 0.2 wt%, from about 5 wt% to about 0.3 wt%, from about 4 wt% to about 0.3 wt%, from about 3 wt% to about 0.3 wt%, from about 2 wt% to about 0.3 wt%, from about 1 wt% to about 0.3 wt%, from about 0.5 wt% to about 0.3 wt%, from about 0.4 wt% to about 0.3 wt%, from about 5 wt% to about 0.4 wt%, from about 4 wt% to about 0.4 wt%, from about 3 wt% to about 0.4 wt%, from about 2 wt% to about 0.4 wt%, from about 1 wt% to about 0.4 wt%, from about 0.5 wt% to about 0.4 wt%, from about 5 wt% to about 0.5 wt%, from about 4 wt% to about 0.5 wt%, from about 3 wt% to about 0.5 wt%, from about 2 wt% to about 0.5 wt%, from about 1 wt% to about 0.5 wt%, from about 5 wt% to about 1 wt%, from about 4 wt% to about 1 wt%, from about 3 wt% to about 1 wt%, from about 2 wt% to about 1 wt%, from about 5 wt% to about 2 wt%, from about 4 wt% to about 2 wt%, from about 3 wt% to about 2 wt%, from about 5 wt% to about 3 wt%, from about 4 wt% to about 3 wt%, from about 5 wt% to about 4 wt%, or adding the polyacrylamide-based copolymer of the present disclosure of about 0.005 wt%, about 0.01 wt%, or about 0.1 wt% to the formulation. In some embodiments, the method includes adding from about 0.0005 wt% to about 5 wt% of the polyacrylamide-based copolymer of the present disclosure to the formulation. In some embodiments, the method is about 0.Including adding from about 0.01 wt% to about 1 wt% of the polyacrylamide-based copolymer of the present disclosure to the formulation. In some embodiments, the formulation includes a biomolecule. In some embodiments, the formulation includes a protein. In some aspects, the formulation includes a lipid-based vehicle. In some embodiments, the lipid-based vehicle includes liposomes, lipid nanoparticles, polymerosomes or micelles.

[0086] 3.2.1. Other uses of polyacrylamide-based copolymers The polyacrylamide-based copolymers of the present disclosure can be used in any composition or formulation in which a surfactant is used. For example, the copolymers of the present disclosure can be used in applications including, but not limited to, cosmetics, hair products, lotions, foods, nutritional products, pigments, and inks. In some embodiments, the copolymer is used to improve texture or mouthfeel. In some embodiments, the copolymer is used in biopharmaceutical compositions for animal or veterinary use. In some embodiments, the presence of the copolymer improves the stability of the formulation.

[0087] 3.3. Definitions Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Methods and materials for use in this application are described herein. In some aspects of the present disclosure, other suitable methods and materials known in the art are also used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. When trade names are used herein, unless otherwise indicated by context, the trade names include product formulations, generics, and pharmaceutical active ingredients of the named products.

[0088] The terms "subject" and "patient" are used interchangeably. A subject can be a mammal such as a non - primate (e.g., cow, pig, horse, cat, dog, goat, rabbit, rat, mouse, etc.) or a primate (e.g., monkey, ape, and human), such as a human. In certain embodiments, the subject is a mammal, such as a human, diagnosed with a disease or disorder provided herein. In another embodiment, the subject is a mammal, such as a human, at risk of developing a disease or disorder provided herein. In certain embodiments, the subject is a human.

[0089] The terms "plurality of treatments" and "treatment" are used in the broadest sense understood in the clinical field.

[0090] In this disclosure, the terms "a", "an", and "the" are used to include one or more unless the context clearly dictates otherwise. The term "or" is used to refer to a non - exclusive "or" unless otherwise specified. The statement "at least one of A and B" is synonymous with "A, B, or A and B". Further, it should be understood that the expressions or terms used in this disclosure are for illustrative purposes only and not for purposes of limitation, unless otherwise defined. The use of section headings is intended to assist in reading the document and should not be construed as limiting. Information related to a section heading may occur within or outside of that particular section.

[0091] Values expressed in a range format should be interpreted flexibly to include not only the numerical values explicitly listed as the limits of the range, but also all individual numerical values or sub-ranges subsumed within that range as if each numerical value and sub-range were explicitly listed. For example, a range of "about 0.1% to about 5%" or "about 0.1% to 5%" should be interpreted to include not only about 0.1% to about 5%, but also the individual values (e.g., 1%, 2%, 3%, and 4%) and sub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the indicated range. The recitation "about X to Y" is synonymous with "about X to about Y" unless otherwise indicated. Similarly, the recitation "about X, Y, or about Z" is synonymous with "about X, about Y, or about Z" unless otherwise indicated.

[0092] As used in this disclosure, the term "about" can allow for a value or range of values within 5% of the recited value or the limits of the recited range, or some degree of variation.

[0093] In the methods described in this disclosure, operations can be performed in any order, except where an explicit temporal or operational sequence is recited. Further, unless the explicit claim language specifies otherwise, the recited operations can be performed simultaneously. For example, the claimed operations of performing X and performing Y can be performed simultaneously within a single operation, and the resulting process falls within the literal scope of the claimed process.

[0094] The term "polymer" refers to a substance or material composed of repeating monomer subunits.

[0095] As used herein, the term "acrylamide monomer" refers to monomer species having an acrylamide functional group. The term "acrylamide monomer" includes not only monomer acrylamide but also derivatives of monomer acrylamide. Examples of acrylamide monomers include, but are not limited to, acrylamide (AM), N-(3-methoxypropyl)acrylamide (MPAM), 4-acryloylmorpholine (MORPH), N,N-dimethylacrylamide (DMA), N-hydroxyethylacrylamide (HEAM), N-[tris(hydroxymethyl)-methyl]acrylamide (TRI), 2-acrylamido-2-methylpropanesulfonic acid (AMP), (3-acrylamidopropyl)trimethylammonium chloride (TMA), N-isopropylacrylamide (NIP), N,N-diethylacrylamide (DEA), N-tert-butylacrylamide (TBA), and N-phenylacrylamide (PHE).

[0096] The term "polyacrylamide-based copolymer" refers to a polymer formed from the polymerization of two or more monomer species, at least one of the monomer species having an acrylamide functional group (acrylamide monomer), and the monomers being structurally different. In some embodiments, the polyacrylamide-based copolymer is formed from the polymerization of two structurally different acrylamide monomers (two structurally different monomers each having an acrylamide functional group). The resulting copolymer can be an alternating copolymer in which the monomer species are linked alternately, a random copolymer in which the monomer species are linked to each other within the polymer chain without a defined pattern, a block copolymer in which a polymer block of one monomer species is linked to a polymer block composed of another monomer species, or a graft copolymer in which the main polymer chain consists of one monomer species and a polymer block of another monomer species is linked to the main polymer chain as a side branch. In some embodiments, the polyacrylamide-based copolymers of the present disclosure are formed from the polymerization of a water-soluble carrier monomer and a functional dopant monomer. In some embodiments, the polyacrylamide-based copolymers of the present disclosure are random copolymers.

[0097] As defined herein, the term "water-soluble carrier monomer" refers to acrylamide monomer species that are water-soluble species within a polyacrylamide-based copolymer. In some embodiments, the water-soluble carrier monomer is the main species within the polyacrylamide-based copolymer. In some embodiments, the water-soluble carrier monomer imparts water solubility to the copolymer. In some embodiments, the water-soluble carrier monomer within the polyacrylamide-based copolymer provides an inert barrier at the interface of the aqueous formulation to prevent protein-protein interactions. In some embodiments, the interface is an air-water interface. In some embodiments, the interface is an enclosure-water interface including, but not limited to, a glass-water interface, a rubber-water interface, a plastic-water interface, or a metal-water interface. In some embodiments, the interface is an oil-water interface. In some embodiments, the interface is the interface between a liquid and a tube. In some embodiments, the interface is the interface between a liquid and a catheter. In some embodiments, the enclosure-water interface is within a pump system. In some embodiments, the enclosure-water interface is within a closed-loop system. In some embodiments, the water-soluble carrier monomer is nonionic. Examples of water-soluble carrier monomers include, but are not limited to, acrylamide (AM), N-(3-methoxypropyl)acrylamide (MPAM), 4-acryloylmorpholine (MORPH), N,N-dimethylacrylamide (DMA), and N-hydroxyethylacrylamide (HEAM).

[0098] As used herein, the term "functional dopant monomer" refers to acrylamide monomer species having physicochemical properties different from those of the water-soluble carrier monomer (e.g., hydrophobicity, charge). In some embodiments, the functional dopant monomer in the polyacrylamide-based copolymer promotes the association of the polymer with the interface. Such interfaces can include, but are not limited to, polymer-air-water interface interactions, polymer-protein interactions, polymer-peptide interactions, polymer-micelle interactions, polymer-liposome interactions, and polymer-lipid nanoparticle interactions. The functional dopant monomer can act as a stabilizing moiety to facilitate interactions with biomolecules, such as proteins, peptides, antibodies, antibody-drug conjugates, nucleic acids, lipid particles, and combinations thereof (e.g., to prevent aggregation of biomolecules). Based on their chemical composition, the functional dopant monomer can be further classified into hydrogen-bonding monomers, ionic monomers, hydrophobic monomers, and aromatic monomers. Typically, the functional dopant monomer is copolymerized at a lower weight percentage compared to the water-soluble carrier monomer.

[0099] The term "polymerization" refers to the process by which monomer molecules undergo a chemical reaction to form polymer chains or three-dimensional networks. For example, different types of polymerization reactions are known in the art, such as addition (chain reaction) polymerization, condensation polymerization, ring-opening polymerization, free radical polymerization, controlled radical polymerization, atom transfer radical polymerization (ATRP), single electron transfer living radical polymerization (SET-LRP), reversible addition-fragmentation chain transfer (RAFT) polymerization, nitroxide-mediated polymerization (NMP), and emulsion polymerization. In some embodiments, the copolymers of the present disclosure are prepared using RAFT polymerization.

[0100] The term "degree of polymerization" (DP) refers to the number of monomer units in a polymer. This is calculated by dividing the average molecular weight of the polymer sample by the molecular weight of the monomer. As defined herein, the average molecular weight of a polymer can be represented by the number average molecular weight (Mn), weight average molecular weight (Mw), Z average molecular weight (Mz), or the molecular weight at the peak maximum of the molecular weight distribution curve (Mp). The average molecular weight of a polymer can be determined by various analytical characterization techniques known to those skilled in the art, such as gel permeation chromatography (GPC), static light scattering (SLS) analysis, multi-angle laser light scattering (MALLS) analysis, nuclear magnetic resonance spectroscopy (NMR), intrinsic viscosity measurement (IV), melt flow index (MFI), and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), and combinations thereof. The degree of polymerization can also be determined experimentally using appropriate analytical methods known in the art, such as nuclear magnetic spectroscopy (NMR), Fourier transform infrared spectroscopy (FT-IR), and Raman spectroscopy.

[0101] The term "amphiphilic" refers to a chemical substance having both hydrophilic (water-loving, polar) and lipophilic (fat-loving, non-polar) properties. Common amphiphilic compounds include detergents, soaps, surfactants, lipoproteins, and phospholipids. In some embodiments, the amphiphilic substance is a charged species. In some embodiments, the amphiphilic substance is a neutral species.

[0102] As used herein, "lipid-based vehicle," refers to a structure having a protective outer layer of lipid that can be used as a drug delivery vehicle. For example, a lipid-based vehicle can be used to encapsulate and transport a payload (e.g., a therapeutic agent) to a biological target. Examples of lipid-based vehicles include, but are not limited to, liposomes, micelles, polymerosomes, and lipid nanoparticles.

[0103] As used herein, "biomolecule" refers to molecules such as proteins, nucleic acids, polysaccharides, and lipids.

[0104] The term "protein" is defined as a class of macromolecules that contain long chains of one or more amino acids. A wide variety of proteins may be considered to belong to a protein family based on having similar structural features, having a particular biological function, and / or being associated with a particular microorganism, particularly a disease-causing microorganism. Such proteins include, for example, antibodies (immunoglobulins), cytokines, chemokines, enzymes, hormones, vaccine antigens, cancer antigens, adjuvants, nutritional markers, and tissue-specific antigens.

[0105] As used herein, the term "nucleic acid" includes deoxyribonucleic acid (DNA), ribonucleic acid (RNA), messenger RNA (mRNA), small interfering RNA (siRNA), short hairpin RNA (shRNA), and microRNA (miRNA).

[0106] The term "antibody" refers to large Y-shaped proteins produced by the immune system to identify and neutralize foreign substances such as pathogenic bacteria and viruses. The term "antibody" includes monoclonal antibodies (e.g., full-length or intact monoclonal antibodies), polyclonal antibodies, multivalent antibodies, multispecific antibodies (e.g., bispecific or trispecific antibodies as long as they exhibit the desired biological activity), and may also include specific antibody fragments. Antibodies can be human, humanized and / or affinity matured. An "antibody fragment" contains only a portion of an intact antibody, and in certain embodiments, the portion retains at least one, typically most or all, of the functions normally associated with that portion when present in the intact antibody. In one embodiment, the antibody fragment contains the antigen-binding site of the intact antibody and thus retains the ability to bind to an antigen. In another embodiment, an antibody fragment, e.g., one containing an Fc region, retains at least one of the biological functions normally associated with the Fc region when present in the intact antibody, such as FcRn binding, antibody half-life regulation, ADCC function, and complement binding. In one embodiment, the antibody fragment is a monovalent antibody having an in vivo half-life substantially similar to that of the intact antibody. For example, such an antibody fragment may contain an antigen-binding arm linked to an Fc sequence that can confer in vivo stability to the fragment.

[0107] The term "aggregation" refers to the formation of higher molecular weight amorphous species by non-covalent attachment ("clumping") of smaller species. The aggregation process can be irreversible or reversible. Many biomolecules and synthetic molecules, including proteins, peptides, lipid particles, nucleic acids, inorganic nanoparticles, and organic nanoparticles (e.g., micelles, lipid nanoparticles, liposomes, polymerosomes) that may further contain encapsulated species, can undergo aggregation.

[0108] In the case of protein aggregation, the formation of protein aggregates can be due to the intrinsic disordered nature of the protein or misfolding of the protein molecule, resulting in the exposure of hydrophobic residues that are normally buried inside the three-dimensional structure of the protein and on the surface. Due to the hydrophobic effect, the exposed hydrophobic portions of misfolded proteins tend to interact with other misfolded protein molecules to shield the exposed hydrophobic surfaces, which can lead to protein aggregation.

[0109] Some biomolecules are "more prone to aggregation" than others. For example, the amino acid sequence and overall three-dimensional structure of a protein are related to its susceptibility to aggregation. For example, transmembrane proteins tend to aggregate (or are more prone to aggregation) than non-membrane proteins, especially when recombinantly expressed without using stabilizers. Proteins exposed to conditions beyond physiological conditions (37 °C, neutral pH, isotonicity) may also be more prone to aggregation than when they are in their native environment. Stress conditions such as temperature fluctuations, light, mechanical perturbations (e.g., shaking), surfaces, ultrasonic vibrations, pH changes, and changes in ionic strength can affect the stability of proteins and induce aggregation. Protein aggregation can lead to the formation of subvisible or visible particles (i.e., precipitates). The extent of subvisible protein aggregation can be measured by various analytical methods known in the art, such as size exclusion chromatography (SEC), gel electrophoresis, asymmetric flow field-flow fractionation (AF4), analytical ultracentrifugation (AUC), mass spectrometry (MS), optical microscopy, fluorescence microscopy, dynamic light scattering (DLS), multi-angle laser light scattering (MALLS), flow imaging, turbidimetry / nephelometry, and transmittance measurements.

[0110] As used herein, the term "decrease in aggregation" of a biomolecule or lipid-based vehicle includes all forms of reducing aggregation. The degree or amount of aggregation observed (e.g., in a composition) can be reduced as compared to a composition of the same biomolecule or lipid-based vehicle in the absence of the polyacrylamide-based copolymer of the present disclosure. Thus, "decrease in aggregation" includes no observable aggregation or a decrease in the amount of aggregation (e.g., a reduced level of aggregated protein). Thus, the amount of aggregates present in a composition is at least about 10 mol%, about 20 mol%, about 30 mol%, about 40 mol%, about 50 mol%, about 60 mol%, about 70 mol%, about 80 mol%, about 90 mol%, or about 100 mol% less as compared to the amount of aggregates of the same biomolecule or lipid-based vehicle in the absence of the polyacrylamide-based copolymer. Aggregation can be measured by any method known in the art including, but not limited to, size exclusion chromatography (SEC), gel electrophoresis, asymmetric flow field-flow fractionation (AF4), analytical ultracentrifugation (AUC), mass spectrometry (MS), light microscopy, fluorescence microscopy, dynamic light scattering (DLS), multi-angle laser light scattering (MALLS), flow imaging, turbidimetry / nephelometry, and transmittance measurements.

[0111] As used herein, the term "increase in stability," when referring to a formulation containing a biomolecule or lipid-based vehicle, refers to a measurable decrease in the amount of aggregation over a period of time under test or fixed storage conditions as compared to the amount of aggregates of the same biomolecule or lipid-based vehicle in the absence of the polyacrylamide-based copolymer.

[0112] As used herein, the terms "aggregated protein" or "protein aggregate" refer to an aggregate of proteins that are disordered or misfolded and grouped together. Aggregates can be soluble or insoluble. Protein aggregates include, but are not limited to, inclusion bodies, soluble and insoluble precipitates, soluble non-native oligomers, gels, protofibrils, films, filaments, protofibrils, amyloid deposits, amyloid protofibrils, plaques, and dispersed non-native intracellular oligomers. In some embodiments, the proteins of the protein aggregates are soluble precursors prior to their aggregation. In compositions containing the polyacrylamide-based copolymers of the present disclosure, protein aggregation can be prevented. The compositions containing the polyacrylamide-based copolymers of the present disclosure can also reduce protein aggregation as compared to compositions containing the same proteins that do not contain the polyacrylamide-based copolymers of the present disclosure. Thus, the polyacrylamide-based copolymers can reduce or prevent protein aggregation.

[0113] 4. Additional Embodiments The present disclosure is further illustrated by the following non-limiting clauses. Clause 1. A water-soluble carrier monomer selected from N-(3-methoxypropyl)acrylamide (MPAM), 4-acryloylmorpholine (MORPH), N,N-dimethylacrylamide (DMA), N-hydroxyethylacrylamide (HEAM), and acrylamide (AM), and combinations thereof, and A functional dopant monomer containing N,N-diethylacrylamide (DEA) A polyacrylamide-based copolymer comprising the same. Clause 2. The copolymer according to Clause 1, wherein the water-soluble carrier monomer is selected from N-(3-methoxypropyl)acrylamide (MPAM) and 4-acryloylmorpholine (MORPH). Clause 3. The copolymer according to Clause 1 or 2, wherein the weight percentage (wt%) of the water-soluble carrier monomer is from 70% to 98%. Item 4. The copolymer according to any one of Items 1 to 3, wherein the weight percentage (% by weight) of N,N - diethylacrylamide (DEA) is from 2% to 30%. Item 5. The copolymer according to Item 4, wherein the weight percentage (% by weight) of N,N - diethylacrylamide (DEA) is from 2% to 16%. Item 6. The copolymer according to any one of Items 2 to 5, wherein the water - soluble carrier monomer is MORPH. Item 7. The copolymer according to any one of Items 2 to 5, wherein the water - soluble carrier monomer is MPAM. Item 8. The copolymer according to any one of Items 1 to 7, further comprising a functional dopant monomer selected from N - [tris(hydroxymethyl)methyl]acrylamide (TRI), 2 - acrylamido - 2 - methylpropane sulfonic acid (AMP), (3 - acrylamidopropyl)trimethylammonium chloride (TMA), N - isopropylacrylamide (NIP), N - tert - butylacrylamide (TBA), and N - phenylacrylamide (PHE), and combinations thereof. Item 9. The copolymer according to any one of Items 1 to 8, wherein the degree of polymerization is from 10 to 500. Item 10. The copolymer according to Item 9, wherein the degree of polymerization is from 20 to 200. Item 11. The copolymer according to Item 10, wherein the degree of polymerization is 50. Item 12. The copolymer according to any one of Items 1 to 11, wherein the molecular weight of the copolymer is from 1,000 to 40,000 g / mol. Item 13. The copolymer according to Item 12, wherein the molecular weight of the copolymer is from 2,000 to 10,000 g / mol. Item 14. The copolymer according to Item 13, wherein the molecular weight of the copolymer is from 4,000 to 6,000 g / mol. Item 15. The copolymer according to any one of Items 1 to 14, wherein the copolymer is amphiphilic. Item 16. A water - soluble carrier monomer containing an acrylamide - reactive moiety, A functional dopant monomer containing N,N - diethylacrylamide (DEA) and a polyacrylamide - based copolymer containing wherein the weight percentage (wt%) of the water - soluble carrier monomer is from 70% to 98%, the weight percentage (wt%) of the functional dopant monomer is from 2% to 30%, the average molecular weight (Mn) of the polyacrylamide - based copolymer is from about 1,000 g / mol to about 30,000 g / mol, and the degree of polymerization is from about 10 to about 250, a polyacrylamide - based copolymer. Article 17. The copolymer according to Article 16, wherein the water - soluble carrier monomer is selected from N - (3 - methoxypropyl) acrylamide (MPAM), 4 - acryloylmorpholine (MORPH), N,N - dimethylacrylamide (DMA), N - hydroxyethylacrylamide (HEAM), and acrylamide (AM). Article 18. The copolymer according to Article 17, wherein the water - soluble carrier monomer is selected from N - (3 - methoxypropyl) acrylamide (MPAM) and 4 - acryloylmorpholine (MORPH). Article 19. The copolymer according to any one of Articles 16 - 18, wherein the functional dopant monomer further comprises a monomer selected from N - [tris (hydroxymethyl) methyl] acrylamide (TRI), 2 - acrylamido - 2 - methylpropane sulfonic acid (AMP), (3 - acrylamidopropyl) trimethylammonium chloride (TMA), N - isopropylacrylamide (NIP), N - tert - butylacrylamide (TBA), and N - phenylacrylamide (PHE), and combinations thereof. Article 20. A composition comprising the copolymer according to any one of Articles 1 - 19 and a pharmaceutically acceptable excipient. Article 21. A composition comprising the copolymer according to any one of Articles 1 - 19, which is a cosmetic, hair product, lotion, food, veterinary product, or nutritional product. Item 22. A composition comprising the copolymer according to any one of Items 1 to 19 and a protein. Item 23. The composition according to Item 22, wherein the protein is a protein that easily aggregates in an aqueous medium. Item 24. The composition according to any one of Items 20 to 23, wherein the copolymer concentration is from 0.1% by weight to about 10% by weight of the composition. Item 25. The composition according to Item 24, wherein the copolymer concentration is 0.5% by weight or about 5% by weight of the composition. Item 26. The composition according to Item 25, wherein the copolymer concentration is 1% by weight of the composition. Item 27. The composition according to any one of Items 20 to 26, comprising one or more of an aqueous buffer, an isotonicity adjusting agent, and a preservative. Item 28. The composition according to any one of Items 20 to 27, wherein the composition is an aqueous composition. Item 29. The composition according to Item 28, wherein the pH of the composition is from 4 to 9. Item 30. The composition according to Item 28, wherein the pH of the composition is from about 6 to about 8. Item 31. A method for increasing the thermal stability of the protein preparation, comprising adding from 0.0005% by weight to 5% by weight of the copolymer according to any one of Items 1 to 19 to the protein preparation. Item 32. A method for increasing the stability of the protein preparation, comprising adding from about 0.005% by weight to about 5% by weight of the copolymer according to any one of Items 1 to 19 to the protein preparation. Item 33. A method for reducing the aggregation rate of a protein in an aqueous composition, comprising adding from 0.005% by weight to 5% by weight of the copolymer according to any one of Items 1 to 19 to the protein preparation. Item 34. The method according to any one of Items 31 to 33, wherein the protein is a protein that aggregates in an aqueous medium.

Example

[0114] 5. Example The examples in this section are provided by way of illustration and not limitation. It should be understood that the examples can represent only some embodiments, and the following examples are illustrative and not limiting. All substituents are as defined previously unless otherwise specified. Reagents and starting materials are readily available to those skilled in the art. The specific synthetic steps for each of the described routes can be combined in different ways or combined with steps from different schemes to prepare the compounds described herein.

[0115] 5.1. Example 1 - Synthesis of Polyacrylamide - based Copolymers Synthesis of the First Copolymer Library by Automated Parallel Synthesis: The copolymerization of the carrier and the dopant is carried out using RAFT polymerization ([Total Monomers] / [CTA]=50, [CTA] / [AIBN]=0.2). MPAM, MORPH, and DMA carrier monomers copolymerized with the DEA dopant monomer are polymerized in DMF using 2 - CPDT or BM1433 as CTA and AIBN as the initiator. The molar concentration of the total vinyl monomers is maintained at 2.72 M (MPAM copolymerization), 2.86 M (MORPH copolymerization), and 3.84 M (DMA copolymerization) such that the homopolymerization of the carrier monomer in DMF is carried out at a constant 40 wt%. The HEAM carrier monomer copolymerized with the DEA dopant monomer is polymerized in a DMF / EtOH mixture using 2 - CPDT or BM1433 as CTA and AIBN as the initiator. The molar concentration of all vinyl monomers is maintained at 2.58 M (HEAM copolymerization) such that the homopolymerization of HEAM in DMF is carried out at a constant 30 wt%. The AM carrier monomer copolymerized with the DEA dopant monomer is polymerized in a DMF / water mixture using BM1433 as CTA and ACVA as the initiator. The molar concentration of all vinyl monomers is maintained at 4.05 M (AM copolymerization) such that the homopolymerization of AM in DMF is carried out at a constant 30 wt%.

[0116] Stock Solutions: Reaction mixtures were prepared by combining (i) the carrier, (ii) the dopant, and (iii) the CTA and the initiator.

[0117] Parallel synthesis of polyacrylamide-based copolymers (also called AC / DC excipients) is carried out on a Chemspeed Swing XL automated synthesis robot equipped with a 4-Needle Head tool and an iSynth reactor. The reactions are performed in 8 mL disposable iSynth reactor vials. All aspirated and dispensed reagent solutions are carried out using a 4-Needle Head tool equipped with 2×10 mL and 2×1 mL syringes with septum piercing needles, with both 1 mL and 10 mL syringes. All solvent series are prepared using 60 mL (6 strokes of syringe volume) of degassed DMF. The typical aspiration and dispensing rates of the reagents are 10 mL / min for both 1 mL syringes. During aspiration using the 4-Needle Head tool, a 50 μL air gap and 50 μL additional volume are used for the 1 mL syringe, and a 50 μL air gap and 100 μL additional volume are used for the 10 mL syringe. After each reagent dispensing operation, the inner and outer volumes of the priming solvent for the 1 mL syringe are 3 mL, and for the 10 mL syringe are 20 mL, and the needles and solvent series are rinsed. The DMF reservoir is degassed by continuous nitrogen sparging. All stock solutions are prepared in septum-capped reagent vials and degassed by sparging with argon for 15 minutes before transferring to Chemspeed. The atmosphere inside Chemspeed is reduced to <1% oxygen by purging with nitrogen with the exhaust port closed. The reactor vials are exposed to a nitrogen stream until the start of the reaction. The calculated aliquots of stock solutions and solvents are transferred to the reactor via an automated liquid handling system. After dispensing, the reaction vials are manually sealed in an inert atmosphere, removed from Chemspeed, manually shaken to combine the reagents, heated in an oven at 65 °C for 24 hours, then the reaction vials are cooled to room temperature and exposed to air.

[0118] The procedure for removing the CTA Z group from the AC / DC excipient containing MORPH, DMA, HEAM, and MPAM copolymers is adapted from the literature. Dilute the reaction vial to 6 mL with DMF. Add LPO (2 equivalents) and AIBN (20 equivalents) to the reaction mixture and seal it with a cap using a PTFE seal. Flush the reaction mixture with nitrogen gas for 10 minutes while heating at 90 °C, followed by heating at 90 °C for 12 hours. The procedure for removing the CTA Z group from the AC / DC excipient containing the AM copolymer is adapted from the literature. Dilute the reaction vial to 5 mL with miliQ water. H 2 O 2 (20 equivalents) is added to the reaction vial, sealed, and heated at 60 °C for 12 hours. The resulting copolymer is isolated by precipitation as outlined below.

[0119] Precipitate the AC / DC excipient synthesized using the carriers of AM and HEAM from acetone twice. Precipitate the AC / DC excipient synthesized with DMA and MORPH from diethyl ether twice. Precipitate the AC / DC excipient synthesized with MPAM from a mixture of diethyl ether and hexane (in a ratio of 3:1) twice. Determine the number (Mn) average molecular weight, weight (Mw) average molecular weight, and dispersity of the AC / DC excipients containing MORPH, MPAM, DMA, and HEAM using SEC in DMF containing poly(ethylene glycol) standards. The Mn, Mw, and dispersity of the AC / DC excipients containing AM are determined using aqueous SEC-MALLS.

[0120] Synthesis of the second polymer library: A typical procedure for synthesizing the MORPH-DEA AC / DC excipient is as follows and is nearly identical for all other carrier / dopant combinations, varying only the choice and concentration of the carrier / dopant. In an 8 mL scintillation vial equipped with a PTFE septum, combine MORPH (41.5 equivalents), DEA (8.5 equivalents), 2CPDT (1 equivalent), and AIBN (0.2 equivalent), and dilute with DMF to a total volume of 2.25 mL (33.3 weight / volume vinyl monomer concentration). Flow nitrogen gas through the reaction mixture for 10 minutes and then heat at 65 °C for 12 hours. To remove the Z-terminus of the resulting polymer, add AIBN (20 equivalents) and LPO (2 equivalents) to the reaction mixture, then flow nitrogen gas through this for 10 minutes and heat at 90 °C for 12 hours. Removal of the Z-group is confirmed by the ratio of refractive index to UV (λ = 310 nm) intensity in SEC analysis. Precipitate the resulting polymer three times from ether and dry overnight under vacuum. Determine the resulting composition and molecular weight by 1 1H NMR spectroscopy and SEC using poly(ethylene glycol) standards.

[0121] Copolymer molecular weight characterization: The Mn, Mw, and dispersity of copolymers with HEAM, DMA, MPAM, and MORPH carrier monomers are determined by SEC performed with poly(ethylene glycol) standards (American Polymer Standards Corporation) after passing through two size exclusion chromatography columns.

[0122] The Mn, Mw, and polydispersity of the copolymer with AM are determined by SEC-MALLS after passing through a size exclusion chromatography column in a mobile phase of phosphate buffered saline containing 300 ppm sodium azide. Detection is performed using an Optilab T-rEX (Wyatt Technology Corporation) refractive index detector operating at 658 nm and a TREOS II light scattering detector (Wyatt Technology Corporation) operating at 659 nm. The dn / dc value of the AM copolymer is assumed to be 0.185 in this medium.

[0123] Method for determining experimental VM weight % values: Handling of viscous monomers (HEAM, MPAM, MORPH) by Chemspeed can result in monomer loadings different from the target monomer loadings for selective copolymerization during the initial AC / DC copolymer library synthesis. The experimental weight percent is estimated from the peak molecular weight (Mp) of the SEC trace. Since only the viscous monomers are affected, changes in Mp result from inappropriate addition of the carrier monomer (except for small changes in the osmotic volume in dopant monomers with different weight percentages compared to the carrier). Therefore, for a given carrier / dopant pair, Mp,max is determined to calculate the experimental weight percent.

[0124] High-Throughput Synthesis of Polyacrylamide Libraries: Libraries of AC / DC excipients are combinatorially synthesized by the statistical copolymerization of a water-soluble carrier monomer and a functional dopant DEA monomer. The carrier monomer is the major species and is responsible for both maintaining solubility and providing an inert barrier to prevent protein–protein interactions. The functional dopant can copolymerize at a lower weight percentage and is statistically incorporated throughout the resulting copolymer. The dopant DEA is selected by design to promote either polymer–interface or polymer–protein interactions. The library targets a degree of polymerization (DP) of 50 for the copolymer. The experimental degree of polymerization (DP) of the carrier monomer is estimated using equation (SI).

Number

[0125] This value is used to estimate the experimental weight percentage (wt%). The experimental weight percentage values of the carrier monomer and the functional dopant monomer for each copolymer are determined using 1 1H NMR and SEC.

[0126] The library is prepared by parallel synthesis using a Chemspeed Swing XL Auto Synthesizer, a liquid handling robot in an inert environment. RAFT polymerization is carried out to provide accurate copolymer stoichiometry, low dispersity, and controlled molecular weight for a wide range of monomers. Polyacrylamide derivatives are used for both carrier and dopant monomers due to the range and availability of commercially available water-soluble monomers (carriers) and functional monomers (dopants) as well as polymer stability. Acrylamide derivatives of monomers often exhibit acute toxicity, while polyacrylamide derivatives demonstrate a high degree of biocompatibility when appropriately purified from their monomer precursors. Furthermore, the reactivity ratios between various acrylamide monomers are close to 1, resulting in copolymers with little to no dopant gradient composition. Carrier monomers include acrylamide (AM), hydroxyethyl acrylamide (HEAM), dimethylacrylamide (DMA), acryloylmorpholine (MORPH), and methoxypropylacrylamide (MPAM) (in increasing order of hydrophobicity) as they are nonionic and water-soluble. The dopant monomer is N,N - diethylacrylamide (DEA), which can be further classified as a hydrophobic monomer based on its chemical composition.

[0127] 6. Equivalents and Incorporation by Reference Although various embodiments of the present disclosure have been specifically shown and described with reference to preferred embodiments and various alternative embodiments, those skilled in the art will understand that various changes in form and detail can be made herein without departing from the spirit and scope of the present disclosure.

[0128] All publications, patents, patent applications, and other documents cited in this application, including U.S. Provisional Application No. 63 / 344,991 and published International Application No. WO 2021 / 211976, are hereby incorporated by reference in their entirety for all purposes as if each individual publication, patent, patent application, or other document was individually indicated to be incorporated by reference for all purposes.

Claims

1. A water-soluble carrier monomer selected from the group consisting of N-(3-methoxypropyl)acrylamide (MPAM), 4-acryloylmorpholine (MORPH), and combinations thereof, Functional dopant monomers containing N,N-diethylacrylamide (DEA) A polyacrylamide copolymer containing, A polyacrylamide copolymer having a number-average molecular weight (Mn) of 1,000 to 40,000 g / mol.

2. The copolymer according to claim 1, wherein the weight percentage (W%) of the water-soluble carrier monomer is 70% to 98%.

3. The copolymer according to claim 1, wherein the weight percentage (W%) of N,N-diethylacrylamide (DEA) is 2% to 30%.

4. The copolymer according to claim 3, wherein the weight percentage (W%) of N,N-diethylacrylamide (DEA) is 2% to 16%.

5. The copolymer according to claim 1, wherein the water-soluble carrier monomer is MORPH.

6. The copolymer according to claim 1, wherein the water-soluble carrier monomer is MPAM.

7. The copolymer according to claim 1, wherein the copolymer further comprises a functional dopant monomer selected from N[tris(hydroxymethyl)-methyl]acrylamide (TRI), 2-acrylamide-2-methylpropanesulfonic acid (AMP), (3-acrylamidopropyl)trimethylammonium chloride (TMA), N-isopropylacrylamide (NIP), N-tert-butylacrylamide (TBA), N-phenylacrylamide (PHE), and combinations thereof.

8. The copolymer according to claim 1, wherein the degree of polymerization measured by 1H nuclear magnetic resonance spectroscopy (NMR) is 10 to 500.

9. The copolymer according to claim 8, wherein the degree of polymerization is 20 to 200.

10. The copolymer according to claim 8, wherein the degree of polymerization is 50 to 100.

11. The copolymer according to claim 1, wherein the number-average molecular weight (Mn) of the copolymer, as measured by size exclusion chromatography (SEC), is 1,000 to 40,000 g / mol.

12. The copolymer according to claim 11, wherein the number-average molecular weight (Mn) of the copolymer is 1,000 to 30,000 g / mol.

13. The copolymer according to claim 11, wherein the number-average molecular weight (Mn) of the copolymer is 2,000 to 10,000 g / mol.

14. The copolymer according to claim 11, wherein the number average molecular weight (Mn) of the copolymer is 3,000 to 7,000 g / mol.

15. The copolymer according to claim 11, wherein the number-average molecular weight (Mn) of the copolymer is 4,000 to 6,000 g / mol.

16. A composition comprising the copolymer according to claim 1 and a pharmaceutically acceptable excipient.

17. A composition comprising the copolymer according to claim 1, which is a cosmetic, hair product, lotion, food, veterinary product, or nutritional product.

18. A composition comprising the copolymer and protein described in Claim 1, wherein the protein is a protein that readily aggregates in an aqueous medium.

19. The composition according to any one of claims 16 to 18, wherein the copolymer concentration is 0.1% to 10% by weight of the composition.

20. The composition according to claim 19, wherein the copolymer concentration is 0.5% to 5% by weight of the composition.

21. The composition according to claim 19, wherein the copolymer concentration is 1% by weight of the composition.

22. A method for increasing the thermal stability of a protein preparation, comprising adding 0.005% to 5% by weight of the copolymer according to claim 1 to the protein preparation.

23. A method for reducing the aggregation rate of a protein in an aqueous composition, comprising adding 0.005% to 5% by weight of the copolymer according to claim 1 to a protein preparation.

24. The method according to claim 22 or 23, wherein the protein is an antibody or a fragment thereof.