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

Abstract

The present invention provides a polyacrylamide-based copolymer excipient containing a water soluble carrier monomer having an acrylamide reactive moiety and a functional dopant monomer, the functional dopant monomer including N, N-diethylacrylamide (DEA). The copolymer can reduce or prevent aggregation of the biomolecules and lipid-based carriers at the hydrophobic interface in the aqueous formulation. Thus, the biomolecules and lipid-based carriers in a formulation containing the polyacrylamide-based copolymer can exhibit higher stability, such as higher thermal stability, compared to the same formulation not containing the polyacrylamide-based copolymer. The polyacrylamide-based copolymers can be used with any biomolecules or lipid-based carriers that are susceptible to aggregation in aqueous media.

Description

1. Introduction

[0001] Many biopharmaceuticals in water-based formulations are susceptible to irreversible aggregation when exposed to high temperatures and / or agitation, requiring careful storage and refrigerated transportation (cold chain) to maintain activity during shelf life. Maintaining the integrity of various aggregation-prone biopharmaceuticals presents a challenge to the pharmaceutical industry, healthcare providers, and people around the world who need such biopharmaceutical treatments. Although the aggregation mechanism varies between various biopharmaceutical compounds, many contain hydrophobic patches or partially denatured to expose hydrophobic segments of the molecule, which drive absorption to interfaces (such as air-water, glass-water, or rubber-water interfaces). The high local concentration of molecules at these interfaces, combined with interface-mediated partial unfolding, triggers the initial nucleation of aggregation events, which paves the way for further aggregation. This tendency to aggregate at the interface increases with increasing formulation concentration, negatively affecting overall formulation stability.

[0002] Excipients are often overlooked as “inactive” ingredients in drug formulations. However, in many drug formulations, excipients play an important role in drug activity, improving the solubility, absorption, and stability of the active ingredient. Methods for stabilizing biopharmaceuticals use excipients that can be easily and stably incorporated into high-concentration commercial formulations without interacting with the therapeutic agent in the bulk formulation, altering its pharmacokinetics, or increasing toxicity.

[0003] Therefore, there is a need for improved biopharmaceutical formulations. 2. Overview

[0004] The present disclosure provides a polyacrylamide-based copolymer excipient containing a water-soluble carrier monomer having an acrylamide-reactive portion and a functional dopant monomer comprising N,N-diethylacrylamide (DEA). The copolymer can reduce or prevent the aggregation of biomolecules and lipid-based carriers at hydrophobic interfaces in aqueous formulations. Therefore, biomolecules and lipid-based carriers in formulations containing polyacrylamide-based copolymers can exhibit higher stability, such as higher thermal stability, compared to the same formulations without the polyacrylamide-based copolymer. Polyacrylamide-based copolymers can be used with any biomolecule or lipid-based carrier that is prone to aggregation in aqueous media, including but 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 copolymers can also be used with lipid-based carriers (including but not limited to liposomes, lipid nanoparticles, polymerosomes, and micelles) that are susceptible to aggregation in aqueous media to prevent or reduce aggregation.

[0006] According to some embodiments, copolymer excipients based on polyacrylamide provide stable biological compound preparations. In some embodiments, these excipients are synthetic copolymers composed of water-soluble carrier monomers (selected to help polymer dissolution) and functional dopant monomers comprising N,N-diethylacrylamide (DEA). It is speculated that DEA monomers promote polymer-interface interactions, reduce biomolecular interactions at the interface, and thus improve the stability of biopharmaceutical preparations. In some embodiments, precise high-throughput synthesis using reverse addition cleavage transfer (RAFT) polymerization is used to generate a library of copolymers based on polyacrylamide including DEA. In the present disclosure, copolymers based on polyacrylamide can provide stable biopharmaceutical preparations that exhibit significantly faster pharmacokinetics than corresponding commercial biopharmaceutical preparations.

[0007] The copolymer excipients described in this application can also be more widely used to improve the thermal stability of biopharmaceutical formulations as simple "drop-in" excipients without changing their biological activity, pharmacokinetics or pharmacodynamics. For example, the copolymer excipients described in this application can be used as drop-in excipients in combination with other formulation methods intended to change or modulate the pharmacokinetics of protein formulations.

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

[0009] As disclosed in the present application, high-throughput controlled radical polymerization techniques can be implemented to generate a library of polyacrylamide-based copolymer excipients, including those containing N,N-diethylacrylamide (DEA) monomers. Non-limiting examples of polyacrylamide-based copolymers provided in the present application include polyacrylamide-based copolymers comprising 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 functional dopant monomers comprising DEA. In some embodiments, the functional dopant monomer comprises DEA and further comprises 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] The present disclosure provides a composition comprising 0.005 wt % to 0.2 wt % of a polyacrylamide-based copolymer comprising 70 wt % to 95 wt % of a MORPH carrier monomer; and 5 wt % to 30 wt % of a DEA dopant monomer.

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

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

[0013] Also provided is a method of increasing the stability of a formulation containing a lipid-based carrier, the method comprising adding to the formulation from about 0.005 wt % to about 5 wt % of the polyacrylamide-based copolymer of the present disclosure.

[0014] Also provided is a method of reducing the aggregation rate of biomolecules in an aqueous composition, the method comprising adding 0.005 wt % to 5 wt % of the polyacrylamide-based copolymer of the present disclosure to the formulation.

[0015] Also provided is a method of reducing the aggregation rate of a protein in an aqueous composition, the method comprising adding to the protein formulation 0.005 wt % to about 5 wt % of the polyacrylamide-based copolymer of the present disclosure.

[0016] Also provided is a method of reducing the aggregation rate of a lipid-based carrier in an aqueous composition, the method comprising adding 0.005 wt % to 5 wt % of the polyacrylamide-based copolymer of the present disclosure to the formulation. 3. Detailed description

[0017] As outlined above, the present disclosure describes a library of copolymers comprising different water-soluble acrylamide monomers and functional dopant monomers, wherein the functional dopant monomers include N,N-diethylacrylamide (DEA) of various weight ratios, molecular weights, and degrees of polymerization. The copolymers disclosed herein can reduce or prevent the aggregation of biomolecules (e.g., proteins and peptides) or lipid-based carriers at hydrophobic interfaces in aqueous formulations, particularly molecules that are prone to aggregation in aqueous media. Proteins and other biomolecules are commonly used to treat a wide variety of diseases and conditions; however, maintaining the stability of such formulations and preventing molecular aggregation is a key challenge facing the biopharmaceutical industry. Proteins, other biomolecules, and lipid-based carriers (including but not limited to liposomes, micelles, polymer bodies, and lipid nanoparticles) may aggregate due to a variety of factors, including thermal 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 bonding, and electrostatic forces. Adsorption of these molecules to various interfaces, especially air-water interfaces, plays a key role in inducing aggregation. These aggregates may trigger immunogenic responses in vivo and lead to the production of anti-therapeutic antibodies.

[0019] The polyacrylamide-based copolymers disclosed herein can reduce or prevent aggregation of biomolecules and lipid-based carriers in aqueous formulations, thereby improving stability. The polyacrylamide-based copolymers can be used with any molecule that is prone to aggregation in aqueous media, including but not limited to proteins, such as antibodies and fragments thereof, cytokines, chemokines, hormones, vaccine antigens, cancer antigens, adjuvants, and combinations thereof.

[0020] The polyacrylamide-based copolymers can also be used with lipid-based carriers (including but not limited to liposomes, lipid nanoparticles, polymersomes, and micelles) that are susceptible to aggregation in aqueous media to prevent or reduce aggregation.

[0021] The polyacrylamide-based copolymers of the present disclosure prevent interactions between molecules (e.g., protein-protein interactions) by providing an inert barrier at the hydrophobic interface of the aqueous formulation, preventing aggregation of proteins, other biomolecules, and lipid-based carriers (e.g., liposomes, lipid nanoparticles, polymer bodies, and micelles). In some embodiments, the hydrophobic interface is an air-water interface. In some embodiments, the hydrophobic interface is a shell-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 in the present disclosure, the subject copolymer can be used as a stabilizer for a preparation containing a biomolecule. In some embodiments, the preparation is a protein preparation. The copolymer based on polyacrylamide can also be used as a stabilizer for a preparation containing a lipid-based carrier (including but not limited to liposomes, lipid nanoparticles, polymer bodies and micelles) that is easy to aggregate in an aqueous medium. In some embodiments, a high-throughput screening of a large combination of acrylamide-based copolymer excipient libraries identified a copolymer comprising N,N-diethylacrylamide (DEA) copolymers. In some embodiments, these copolymers enhance the stability of the preparation without any modification of the molecules in the preparation. For example, the copolymer can enhance the stability of a protein preparation without any protein modification. In some embodiments, a copolymer comprising a water-soluble "carrier" monomer and a functional "doping" monomer acts as a stability excipient to reduce the interaction of biomolecules with an interface (e.g., air-liquid interface). In some embodiments, the biomolecule is a protein. In some embodiments, copolymers comprising a water-soluble "carrier" monomer and a functional "doping" monomer act as a stabilizing excipient to reduce the interaction of the lipid-based carrier with interfaces (eg, air-liquid interfaces).

[0023] Therefore, the present disclosure provides a method for reducing the aggregation of biomolecules or lipid-based carriers, which includes a polyacrylamide-based copolymer excipient disclosed in the present application. In some embodiments, the biomolecule is a protein. In some embodiments, the protein is selected from antibodies and fragments thereof, 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 carrier is a liposome, a micelle, a polymer body or a lipid nanoparticle. The polyacrylamide-based copolymer of the present disclosure comprises a water-soluble carrier monomer containing an acrylamide reactive portion and a functional dopant monomer containing an acrylamide reactive portion. In some embodiments, the copolymer comprises an MPAM or MORPH carrier monomer and a DEA dopant monomer.

[0024] Also provided are methods for increasing the stability of formulations containing biomolecules or lipid-based carriers. In some embodiments, methods for increasing the thermal stability of formulations containing biomolecules or lipid-based carriers are provided. In other embodiments, methods for reducing the aggregation rate of biomolecules or lipid-based carriers in aqueous compositions are provided. The methods include adding a copolymer of the present disclosure to the formulation.

[0025] 3.1 Pharmaceutical Composition

[0026] As summarized above, the present disclosure provides a pharmaceutical composition comprising a polyacrylamide-based copolymer containing a water-soluble carrier monomer and a functional dopant monomer including N,N-diethylacrylamide (DEA). In some embodiments, the polyacrylamide-based copolymer is an amphiphilic copolymer.

[0027] 3.1.1. Polyacrylamide-based copolymers

[0028] The present disclosure provides a copolymer based on polyacrylamide. In some embodiments, the copolymer based on polyacrylamide comprises a water-soluble carrier monomer and a functional dopant monomer, and the functional dopant monomer includes N,N-diethylacrylamide (DEA). In some embodiments, the copolymer based on polyacrylamide is an amphiphilic copolymer.

[0029] In some embodiments, the polyacrylamide-based copolymer comprises a nonionic water-soluble acrylamide monomer and a functional dopant monomer, and the functional dopant monomer is N,N-diethylacrylamide (DEA). In some embodiments, the polyacrylamide-based copolymer further comprises a functional acrylamide dopant monomer selected from the group consisting of 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.

[0030] The polyacrylamide-based copolymers disclosed herein contain water-soluble carrier monomers. In some embodiments, the water-soluble carrier monomers are nonionic. In some embodiments, the water-soluble carrier monomers are selected from N-(3-methoxypropyl) acrylamide (MPAM), 4-acryloyl morpholine (MORPH), N,N-dimethylacrylamide (DMA), N-hydroxyethyl acrylamide (HEAM) and acrylamide (AM) or combinations thereof. In some embodiments, the water-soluble carrier monomers are 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-acryloyl morpholine (MORPH). In some embodiments, the water-soluble carrier monomer is N,N-dimethylacrylamide (DMA). In some embodiments, the water-soluble carrier monomer is N-hydroxyethyl acrylamide (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).

[0031] The polyacrylamide-based copolymer of the present disclosure also includes a functional dopant monomer, which includes N,N-diethylacrylamide (DEA), for example, consisting essentially of DEA or consisting of DEA. In some embodiments, the polyacrylamide-based copolymer also 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 also 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 also includes a hydrogen-bonded 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 comprises 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).

[0032] 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 including N,N-diethylacrylamide (DEA).

[0033] 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).

[0034] 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).

[0035] 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).

[0036] In some embodiments, the water-soluble carrier monomer is N-hydroxyethyl acrylamide (HEAM). In some embodiments, the water-soluble carrier monomer is N-hydroxyethyl acrylamide (HEAM), and the functional dopant monomer is N,N-diethyl acrylamide (DEA).

[0037] 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).

[0038] In some embodiments, the polyacrylamide-based copolymer includes N-(3-methoxypropyl)acrylamide (MPAM) or 4-acryloylmorpholine (MORPH) as a water-soluble carrier monomer, and N,N-diethylacrylamide (DEA) as a functional dopant monomer.

[0039] In some embodiments, the polyacrylamide-based copolymer comprises N,N-dimethylacrylamide (DMA), N-hydroxyethylacrylamide (HEAM), or acrylamide (AM) as a water-soluble carrier monomer, and the functional dopant monomer is N,N-diethylacrylamide (DEA).

[0040] In some embodiments, the amount of 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 a functional dopant monomer, for example, 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 a functional dopant monomer. In some embodiments, the polyacrylamide-based copolymer contains 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 2 to 30 wt % of the functional dopant monomer, for example, 5 to 30 wt %, 10 to 30 wt %, 15 to 30 wt %, 20 to 30 wt %, 25 to 30 wt %, 2 to 25 wt %, 5 to 25 wt %, 10 to 25 wt %, 15 to 25 wt %, 20 to 25 wt %, 2 to 20 wt %, 5 to 20 wt %, 10 to 20 wt %, 15 to 20 wt %, 2 to 15 wt %, 5 to 15 wt %, 10 to 15 wt %, 2 to 10 wt %, 5 to 10 wt % or 2 to 5 wt % of the functional dopant monomer. In some embodiments, the polyacrylamide-based copolymer comprises 2%, 5%, 8%, 10%, 12%, 15%, 18%, 20%, 22%, 25%, 28% or 30% by weight of the functional dopant monomer.In some embodiments, the polyacrylamide-based copolymer comprises 2%, 5%, 8%, 10%, 12%, 15%, 18%, 20%, 22%, 25%, 28%, or 30% by weight of N,N-diethylacrylamide (DEA).

[0041] In some embodiments, the polyacrylamide-based copolymer comprises from about 70% to about 98% by weight of a water-soluble carrier monomer, e.g., from about 75% to about 98% by weight, from about 80% to about 98% by weight, from about 85% to about 98% by weight, from about 90% to about 98% by weight, from about 95% to about 98% by weight, from about 70% to about 95% by weight, from about 75% to about 95% by weight, from about 80% to about 95 ... In some embodiments, the polyacrylamide-based copolymer comprises about 70%, about 72%, about 75%, about 78%, about 80%, about 82%, about 85%, about 88%, about 90%, about 90%, about 85%, about 90%, about 70%, about 85%, about 75%, about 85%, about 80%, about 85%, about 70%, about 80%, about 75%, about 80%, about 85%, about 70%, about 80%, about 75%, about 80%, about 70%, about 80%, about 75%, about 80%, or about 70% to 75% by weight of a water-soluble carrier monomer. In some embodiments, the polyacrylamide-based copolymer comprises about 70%, about 72%, about 75%, about 78%, about 80%, about 82%, about 85%, about 88%, about 90%, about 92%, about 95%, or about 98% by weight of a water-soluble carrier monomer. In some embodiments, the polyacrylamide-based copolymer comprises 70% to 98% by weight of a water-soluble carrier monomer, e.g., 75% to 98%, 80% to 98%, 85% to 98%, 90% to 98%, 95% to 98%, 70% to 95%, 75% to 95%, 80% to 95%, 85% to 95%, 90% to 95%, 70% to 90%, 75% to 90%, 80% to 90%, 85% to 90%, 70% to 85%, 75% to 85%, 80% to 85%, 70% to 80%, 75% to 80%, or 70% to 75% by weight of a water-soluble carrier monomer. In some embodiments, the polyacrylamide-based copolymer comprises 70%, 72%, 75%, 78%, 80%, 82%, 85%, 88%, 90%, 92%, 95%, or 98% by weight of a water-soluble carrier monomer.

[0042] In some embodiments, the polyacrylamide-based copolymer comprises about 70% to about 98% by weight of a water-soluble carrier monomer and about 2% to about 30% by weight of a functional dopant monomer. For example, the polyacrylamide-based copolymer may contain about 70% to about 98% by weight, about 70% to about 95% by weight, about 70% to about 80% by weight, about 80% to about 90% by weight, about 90% to about 98% by weight, about 70% by weight, about 75% by weight, about 80% by weight, about 85% by weight, about 90% by weight, about 95% by weight, or about 98% by weight of a water-soluble carrier monomer, and about 2% to about 30% by weight, about 25% to about 30% by weight, about 5% to about 50% by weight, about 20% to about 25% by weight, about 25% to about 25 ... To about 20 wt%, about 2 wt% to about 5 wt%, about 5 wt% to about 10 wt%, about 10 wt% to about 15 wt%, about 15 wt% to about 20 wt%, about 20 wt% to about 25 wt% or 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 functional dopant monomers.

[0043] In some embodiments, the polyacrylamide-based copolymer comprises 70% to 98% by weight of a water-soluble carrier monomer and 2% to 30% by weight of a functional dopant monomer. For example, the polyacrylamide-based copolymer may contain 70% to 98% by weight, 70% to 95% by weight, 70% to 80% by weight, 80% to 90% by weight, 90% to 98% by weight, 70%, 75%, 80%, 85%, 90%, 95% or 98% by weight of a water-soluble carrier monomer and 2% to 30%, 5% to 25%, 5% to 30% by weight of a functional dopant monomer. % to 20 wt%, 2 wt% to 5 wt%, 5 wt% to 10 wt%, 10 wt% to 15 wt%, 15 wt% to 20 wt%, 20 wt% to 25 wt% or 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 the functional dopant monomer.

[0044] In some embodiments, the copolymer based on polyacrylamide contains about 2% to about 30% by weight of the functional dopant monomer DEA. In some embodiments, the copolymer based on polyacrylamide contains about 5% to about 30% by weight of the functional dopant monomer DEA. In some embodiments, the copolymer based on polyacrylamide contains about 10% to about 28% by weight of the functional dopant monomer DEA. In some embodiments, the copolymer based on polyacrylamide contains about 5% to about 26% by weight of the functional dopant monomer DEA. In some embodiments, the copolymer based on polyacrylamide contains about 5% to about 10% by weight of the functional dopant monomer DEA. In some embodiments, the copolymer based on polyacrylamide contains about 10% to about 15% by weight of the functional dopant monomer DEA. In some embodiments, the copolymer based on polyacrylamide contains about 15% to about 20% by weight of the functional dopant monomer DEA. In some embodiments, the copolymer based on polyacrylamide contains about 20% to about 26% by weight of the functional dopant monomer DEA.

[0045] In some embodiments, the polyacrylamide-based copolymer contains 2 wt % to 30 wt % of the functional dopant monomer DEA, for example, 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 %, or 20 wt % to 26 wt % of the functional dopant monomer DEA.

[0046] In some embodiments, the copolymer based on polyacrylamide comprises MORPH as a water-soluble carrier monomer and about 2% to about 30% by weight of DEA as a functional dopant monomer. In some embodiments, the copolymer comprises MORPH and about 5% to about 30% by weight of DEA. In some embodiments, the copolymer comprises MORPH and about 10% to about 28% by weight of DEA. In some embodiments, the copolymer comprises MORPH and about 5% to about 26% by weight of DEA. In some embodiments, the copolymer comprises MORPH and about 5% to about 10% by weight of DEA. In some embodiments, the copolymer comprises MORPH and about 10% to about 15% by weight of DEA. In some embodiments, the copolymer comprises MORPH and about 15% to about 20% by weight of DEA. In some embodiments, the copolymer comprises MORPH and about 20% to about 25% by weight of DEA. In some embodiments, the copolymer comprises MORPH and about 25% to about 30% by weight of DEA. In some embodiments, the copolymer comprises MORPH and about 20% to about 28% by weight of DEA. In some embodiments, the copolymer comprises MORPH and about 21% by weight of DEA. In some embodiments, the copolymer comprises MORPH and about 22% by weight of DEA. In some embodiments, the copolymer comprises MORPH and about 23% by weight of DEA. In some embodiments, the copolymer comprises MORPH and about 24% by weight of DEA. In some embodiments, the copolymer comprises MORPH and about 25% by weight of DEA.

[0047] In some embodiments, the polyacrylamide-based copolymer comprises MORPH as a water-soluble carrier monomer and 2 to 30 wt % DEA as a functional dopant monomer, such as 5 to 30 wt %, 10 to 28 wt %, 5 to 26 wt %, 5 to 10 wt %, 10 to 15 wt %, 15 to 20 wt %, 20 to 25 wt %, 25 to 30 wt %, or 20 to 28 wt % DEA. In some embodiments, the copolymer comprises MORPH and 21 wt % DEA, such as 22 wt %, 23 wt %, 24 wt %, or 25 wt % DEA.

[0048] In some embodiments, the copolymer based on polyacrylamide comprises MPAM as a water-soluble carrier monomer and about 2 wt % to about 16 wt % of DEA as a functional dopant monomer. In some embodiments, the copolymer based on polyacrylamide comprises MPAM as a water-soluble carrier monomer and about 5 wt % to about 15 wt % of DEA as a functional dopant monomer. In some embodiments, the copolymer based on polyacrylamide comprises MPAM as a water-soluble carrier monomer and about 6 wt % to about 10 wt % of DEA as a functional dopant monomer. In some embodiments, the copolymer based on polyacrylamide comprises MPAM as a water-soluble carrier monomer and about 7 wt % of DEA as a functional dopant monomer. In some embodiments, the copolymer based on polyacrylamide comprises MPAM as a water-soluble carrier monomer and about 8 wt % of DEA as a functional dopant monomer. In some embodiments, the copolymer based on polyacrylamide comprises MPAM as a water-soluble carrier monomer and about 9 wt % of DEA as a functional dopant monomer.

[0049] In some embodiments, the polyacrylamide-based copolymer comprises MPAM as a water-soluble carrier monomer and 2 to 16 wt % DEA, such as 5 to 15 wt % or 6 to 10 wt % DEA as a functional dopant monomer. In some embodiments, the polyacrylamide-based copolymer comprises MPAM as a water-soluble carrier monomer and 7 wt % DEA as a functional dopant monomer, such as 8 or 9 wt % DEA as a functional dopant monomer.

[0050] In some embodiments, the polyacrylamide-based copolymer further comprises NIP, TRI, AMP, TMA, TBA or PHE as a functional dopant monomer. In some embodiments, NIP, AMP, TMA, TBA or PHE functional dopant monomers account for about 2% to about 16% of the weight of the copolymer. In some embodiments, NIP, TRI, AMP, TMA, TBA or PHE functional dopant monomers account for about 5% to about 15% of the weight of the copolymer. In some embodiments, NIP, TRI, AMP, TMA, TBA or PHE functional dopant monomers account for about 6% to about 14% of the weight of the copolymer. In some embodiments, NIP, TRI, AMP, TMA, TBA or PHE functional dopant monomers account for about 12% to about 15% of the weight of the copolymer. In some embodiments, NIP, TRI, AMP, TMA, TBA or PHE functional dopant monomers account for about 2% to about 5% of the weight of the copolymer. In some embodiments, NIP, TRI, AMP, TMA, TBA or PHE functional dopant monomers account for about 5% to about 10% of the weight of the copolymer.

[0051] In some embodiments, the polyacrylamide-based copolymer further comprises NIP, AMP, TMA, TBA or PHE as a functional dopant monomer, and its content is 2% to 16% by weight of the copolymer, for example, 5% to 15%, 6% to 14% or 12% to 15%. In some embodiments, the amount of NIP, TRI, AMP, TMA, TBA or PHE functional dopant monomer is 2% to 5% by weight of the copolymer. In some embodiments, the amount of NIP, TRI, AMP, TMA, TBA or PHE functional dopant monomer is 5% to 10% by weight of the copolymer.

[0052] In some embodiments, the degree of polymerization (DP) of the polyacrylamide-based copolymer is about 10 to about 500, about 20 to about 200, about 50 to about 100, about 100 to about 200, about 200 to about 300, about 300 to about 400, or 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 copolymer has a DP of about 100.

[0053] In some embodiments, the degree of polymerization (DP) of the polyacrylamide-based copolymer is 10 to 500, 20 to 200, 50 to 100, 100 to 200, 200 to 300, 300 to 400, or 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, 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.

[0054] In some embodiments, the molecular weight of the polyacrylamide-based copolymer is from about 1,000 g / mol to about 40,000 g / mol, for example, 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. g / mol, about 1,000 g / mol to about 5,000 g / mol, about 1,000 g / mol to about 4,000 g / mol, about 1,000 g / mol to about 3,000 g / mol, about 3,000 g / mol to about 40,000 g / mol, about 3,000 g / mol to about 35,000 g / mol, about 3,000 g / mol to about 30,000 g / mol, about 3,000 g / mol to about 25,000 g / mol, about 3,000 g / mol to about 20,000 g / mol, about 3,000 g / mol to about 15,000 g / mol, about 3,000 g / mol to about 10,000 g / mol, 1, about 3,000 g / mol to about 7,000 g / mol, about 3,000 g / mol to about 6,000 g / mol, about 3,000 g / mol to about 5,000 g / mol, about 3,000 g / mol to about 4,000 g / mol, about 4,000 g / mol to about 40,000 g / mol, about 4,000 g / mol to about 35,000 g / mol, about 4,000 g / mol to about 30,000 g / mol, about 4,000 g / mol to about 25,000 g / mol, about 4,000 g / mol to about 20,000 g / mol, about 4,000 g / mol to about 15,000 g / mol, about g / mol, about 4,000 g / mol to about 10,000 g / mol, about 4,000 g / mol to about 7,000 g / mol, about 4,000 g / mol to about 6,000 g / mol, about 4,000 g / mol to about 5,000 g / mol, about 5,000 g / mol to about 40,000 g / mol, about 5,000 g / mol to about 35,000 g / mol, about 5,000 g / mol to about 30,000 g / mol, about 5,000 g / mol to about 25,000 g / mol, about 5,000 g / mol to about 20,000 g / mol, about 5,000 g / mol to about 15,000 g / mol, about 5,g / mol, about 5,000 g / mol to about 7,000 g / mol, about 5,000 g / mol to about 6,000 g / mol, about 6,000 g / mol to about 40,000 g / mol, about 6,000 g / mol to about 35,000 g / mol, about 6,000 g / mol to about 30,000 g / mol, about 6,000 g / mol to about 25,000 g / mol, about 6,000 g / mol to about 20,000 g / mol, about 6,000 g / mol to about 15,000 g / mol, about 6,000 g / mol to about 10,000 g / mol, about 6,000 g / mol to about 7,000 g / mol, about 7,000 g / mol to about 40,000 g / mol, about 7,000 g / mol to about 35,000 g / mol, about 7000 g / mol to about 30,000 g / mol, about 7,000 g / mol to about 25,000 g / mol, about 7,000 g / mol to about 20,000 g / mol, about 7,000 g / mol to about 15,000 g / mol, about 7,000 g / mol to about 10,000 g / mol, about 10,000 g / mol to about 40,000 g / mol, about 10,000 g / mol to about 35,000 g / mol, about 7000 g / mol to about 30,000 g / mol, about 7,000 g / mol to about 25,000 g / mol, about 7,000 g / mol to about 20,000 g / mol, about 7,000 g / mol to about 15,000 g / mol, about 7,000 g / mol to about 10,000 g / mol, about 10,000 g / mol to about 40,000 g / mol, about 10,000 g / mol to about 35,000 g / mol g / mol, about 10,000 g / mol to about 30,000 g / mol, about 10,000 g / mol to about 25,000 g / mol, about 10,000 g / mol to about 20,000 g / mol, about 10,000 g / mol to about 15,000 g / mol, about 15,000 g / mol to about 40,000 g / mol, about 15,000 g / mol to about 35,000 g / mol, about 15,000 g / mol to about 30,000 g / mol, about 15,000 g / mol to about 25,000 g / mol, about 15,000 g / mol to about 20,000 g / mol, about 20,000 g / mol g / mol to about 40,000 g / mol, about 20,000 g / mol to about 35,000 g / mol, about 20,000 g / mol to about 30,000 g / mol, about 20,000 g / mol to about 25,000 g / mol, about 25,000 g / mol to about 40,000 g / mol, about 25,000 g / mol to about 35,000 g / mol, about 25,000 g / mol to about 30,000 g / mol, about 30,000 g / mol to about 40,000 g / mol, about 30,000 g / mol to about 35,000 g / mol, or about 35,000 g / mol to about 40,000 g / mol.000g / mol. In some embodiments, the molecular weight of the copolymer is from about 1,000 to about 30,000g / mol. In some embodiments, the molecular weight of the copolymer is from about 10,000 to about 20,000g / mol. In some embodiments, the molecular weight of the copolymer is from about 15,000 to about 20,000g / mol. In some embodiments, the molecular weight of the copolymer is from about 20,000 to about 25,000g / mol. The molecular weight of the copolymer is from about 25,000 to about 30,000g / mol. In some embodiments, the molecular weight of the copolymer is from about 30,000 to about 40,000g / mol. In some embodiments, the molecular weight of the copolymer is from about 2,000 to about 10,000g / mol. In some embodiments, the molecular weight of the copolymer is from about 3,000 to about 7,000g / mol. In some embodiments, the molecular weight of the copolymer is from about 4,000 to about 6,000g / mol. ,

[0055] In some embodiments, the molecular weight of the polyacrylamide-based copolymer is 1,000 g / mol to 40,000 g / mol, for example, 1,000 g / mol to 35,000 g / mol, 1,000 g / mol to 30,000 g / mol, 1,000 g / mol to 25,000 g / mol, 1,000 g / mol to 20,000 g / mol, 1,000 g / mol to 15,000 g / mol, 1,000 g / mol to 10,000 g / mol, 1,000 g / mol to 7,000 g / mol, 1,000 g / mol to 6,000 g / mol, 1,000 g / mol to 5,000 g / mol. 00 g / mol, 1,000 g / mol to 4,000 g / mol, 1,000 g / mol to 3,000 g / mol, 3,000 g / mol to 40,000 g / mol, 3,000 g / mol to 35,000 g / mol, 3,000 g / mol to 30,000 g / mol, 3,000 g / mol to 25,000 g / mol, 3,000 g / mol to 20,000 g / mol, 3,000 g / mol to 15,000 g / mol, 3,000 g / mol to 10,000 g / mol, 3,000 g / mol to 7,000 g / mol, 3,000 g / mol to 6,000 g / mol. 00 g / mol, 3,000 g / mol to 5,000 g / mol, 3,000 g / mol to 4,000 g / mol, 4,000 g / mol to 40,000 g / mol, 4,000 g / mol to 35,000 g / mol, 4,000 g / mol to 30,000 g / mol, 4,000 g / mol to 25,000 g / mol, 4,000 g / mol to 20,000 g / mol, 4,000 g / mol to 15,000 g / mol, 4,000 g / mol to 10,000 g / mol, 4,000 g / mol to 7,000 g / mol, 4,000 g / mol to 6,000 g / mol. 00 g / mol, 4,000 g / mol to 5,000 g / mol, 5,000 g / mol to 40,000 g / mol, 5,000 g / mol to 35,000 g / mol, 5,000 g / mol to 30,000 g / mol, 5,000 g / mol to 25,000 g / mol, 5,000 g / mol to 20,000 g / mol, 5,000 g / mol to 15,000 g / mol, 5,000 g / mol to 10,000 g / mol, 5,000 g / mol to 7,000 g / mol, 5,000 g / mol to 6,000 g / mol, 6,000 g / mol to 40,000 g / mol, 6,000 g / mol to 35,000 g / mol, 6,000 g / mol to 30,000 g / mol, 6,000 g / mol to 25,000 g / mol, 6,000 g / mol to 20,000 g / mol, 6,000 g / mol to 15,000 g / mol, 6,000 g / mol to 10,000 g / mol, 6,000 g / mol to 7,000 g / mol, 7,000 g / mol to 40,000 g / mol, 7,000 g / mol to 35,000 g / mol, 00 g / mol, 7000 g / mol to 30,000 g / mol, 7,000 g / mol to 25,000 g / mol, 7,000 g / mol to 20,000 g / mol, 7,000 g / mol to 15,000 g / mol, 7,000 g / mol to 10,000 g / mol, 10,000 g / mol to 40,000 g / mol, 10,000 g / mol to 35,000 g / mol, 10,000 g / mol to 30,000 g / mol, 10,000 g / mol to 25, 000 g / mol, 10,000 g / mol to 20,000 g / mol, 10,000 g / mol to 15,000 g / mol, 15,000 g / mol to 40,000 g / mol, 15,000 g / mol to 35,000 g / mol, 15,000 g / mol to 30,000 g / mol, 15,000 g / mol to 25,000 g / mol, 15,000 g / mol to 20,000 g / mol, 20,000 g / mol to 40,000 g / mol, 20,000 g / mol to 35,000 g / mol, 15,000 g / mol to 30,000 g / mol, 15,000 g / mol to 25,000 g / mol, 15,000 g / mol to 20,000 g / mol, 20,000 g / mol to 40,000 g / mol, 20,000 g / mol to 35,000 g / mol In some embodiments, the molecular weight of the copolymer is 1,000 to 30,000 g / mol. In some embodiments, the molecular weight of the copolymer is 10,000 to 20,000 g / mol. In some embodiments, the molecular weight of the copolymer is 15,000 to 20,000 g / mol. In some embodiments, the molecular weight of the copolymer is 20,000 to 25,In some embodiments, 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.

[0056] The present disclosure also provides a copolymer based on polyacrylamide, which includes a water-soluble carrier monomer containing an acrylamide reactive portion and a functional dopant monomer containing DEA. In some embodiments, the copolymer based on polyacrylamide includes about 70% to about 98% of a water-soluble carrier monomer with an acrylamide reactive portion and about 2% to about 30% of DEA. In some embodiments, the number average molecular weight (Mn) of the copolymer is about 1,000 g / mol to about 30,000 g / mol. In some embodiments, the degree of polymerization is about 10 to about 250. In some embodiments, the copolymer based on polyacrylamide includes 70% to about 98% of a water-soluble carrier monomer with an acrylamide reactive portion and about 2% to about 30% of DEA. In some embodiments, the number average molecular weight (Mn) of the copolymer is 1,000 g / mol to 30,000 g / mol. In some embodiments, the degree of polymerization is 10 to 250. In some embodiments, the water-soluble carrier monomer is nonionic. In some embodiments, the copolymer is amphiphilic.

[0057] In some embodiments, the polyacrylamide-based copolymer comprises about 70 wt % to about 95 wt % of a water-soluble carrier monomer MORPH and about 5 wt % to about 30 wt % of a functional dopant monomer DEA, wherein the copolymer has a number average molecular weight (Mn) of about 1,000 g / mol to about 10,000 g / mol and a degree of polymerization of about 10 to about 100. In some embodiments, the polyacrylamide-based copolymer comprises about 74 wt % to about 80 wt % of a water-soluble carrier monomer MORPH and about 20 wt % to about 26 wt % of a functional dopant monomer DEA, wherein the copolymer has a number average molecular weight (Mn) of about 1,000 g / mol to about 5,000 g / mol and a degree of polymerization of about 10 to about 50. In some embodiments, the polyacrylamide-based copolymer comprises about 77 wt % of a water-soluble carrier monomer MORPH and about 23 wt % of a functional dopant monomer DEA, wherein the copolymer has a number average molecular weight (Mn) of about 3200 g / mol and a degree of polymerization of about 26.

[0058] In some embodiments, the polyacrylamide-based copolymer comprises 70 wt % to 95 wt % of the water-soluble carrier monomer MORPH and 5 wt % to 30 wt % of the functional dopant monomer DEA, wherein the number average molecular weight (Mn) of the copolymer is 1,000 g / mol to 10,000 g / mol, and the degree of polymerization is 10 to 100. In some embodiments, the polyacrylamide-based copolymer comprises 74 wt % to 80 wt % of the water-soluble carrier monomer MORPH and 20 wt % to 26 wt % of the functional dopant monomer DEA, wherein the number average molecular weight (Mn) of the copolymer is 1,000 g / mol to 5,000 g / mol, and the degree of polymerization is 10 to 50. In some embodiments, the polyacrylamide-based copolymer comprises 77 wt % of the water-soluble carrier monomer MORPH and 23 wt % of the functional dopant monomer DEA, wherein the number average molecular weight (Mn) of the copolymer is 3200 g / mol, and the degree of polymerization is 26.

[0059] In some embodiments, the composition comprises from about 0.001% to about 1% by weight of the polyacrylamide-based copolymer, for example, from about 0.001% to about 0.5%, from about 0.001% to about 0.1%, from about 0.001% to about 0.05%, from about 0.001% to about 0.03%, from about 0.005% to about 1%, from about 0.005% to about 0.5%, from about 0.005% to about 0.1%, from about 0.005% to about 0.05%, from about 0.005% to about 0.03%, from about 0.01% to about 1%, from about 0.01% to about 0.5%, from about 0.01% to about 0.1%, from about 0.01% to about 0.05%, or from about 0.01% to about 0.03% by weight of the polyacrylamide-based copolymer. In some embodiments, the composition comprises about 0.01 wt % of the polyacrylamide-based copolymer.

[0060] In some embodiments, the composition comprises 0.001% to 1% by weight of the polyacrylamide-based copolymer, for example, 0.001% to 0.5%, 0.001% to 0.1%, 0.001% to 0.05%, 0.001% to 0.03%, 0.005% to 1%, 0.005% to 0.5%, 0.005% to 0.1%, 0.005% to 0.05%, 0.005% to 0.03%, 0.01% to 1%, 0.01% to 0.5%, 0.01% to 0.1%, 0.01% to 0.05%, or 0.01% to 0.03% by weight of the polyacrylamide-based copolymer. In some embodiments, the composition comprises 0.01 wt % of the polyacrylamide-based copolymer.

[0061] 3.1.2. Compositions containing polyacrylamide-based copolymers

[0062] Compositions comprising the polyacrylamide-based copolymers of the present disclosure are also provided. In some embodiments, the composition comprises the polyacrylamide-based copolymers of the present disclosure and a pharmaceutically acceptable excipient.

[0063] In some embodiments, the composition comprises a copolymer based on polyacrylamide 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 easily aggregated in an aqueous medium. In some embodiments, the copolymer is uncharged, cationic or anionic. In some embodiments, the copolymer is amphiphilic. In some embodiments, the carrier monomer is a water-soluble substance responsible for maintaining solubility and providing an inert barrier to prevent biomass aggregation, for example, to prevent protein-protein interactions. In some embodiments, the water-soluble carrier monomer is the main substance in a copolymer based on polyacrylamide. In some embodiments, the functional dopant monomer (e.g., DEA) is copolymerized at a lower weight percentage and statistically blended throughout the resulting copolymer. In some embodiments, the dopant promotes polymer-interface interactions or polymer-protein interactions.

[0064] In some embodiments, a composition comprising a polyacrylamide-based copolymer of the present disclosure and a biomolecule or lipid-based carrier has a copolymer concentration of about 0.0001% to about 5% by weight of the composition, for example, about 0.0001% to about 4%, about 0.0001% to about 3%, about 0.0001% to about 2%, about 0.0001% to about 1%, about 0.0001% to about 0.5%, about 0.0001% to about 0.4%, about 0.0001% to about 0.3%, about 0.0001% to about 0.2%, about 0.0001% to about 0.1%, about 0.0001% to about 0.05%, about 0.0001% to about 0.02%, about 0.0001% to about 0.05%, about 0.0001% to about 0.06%, about 0.0001% to about 0.08%, about 0.0001% to about 0.11%, about 0.0001% to about 0.09%, about 0.010% to about 0.011%, about 0.011% to about 0.012%, about 0.011% to about 0.013%, about 0.012% to about 0.014%, about 0.013% to about 0.015%, about 0.014% to about 0.016%, about 0.015% to about 0.017%, .01%, about 0.0001% to about 0.005%, about 0.005% to about 5%, about 0.005% to about 4%, about 0.005% to about 3%, about 0.005% to about 2%, about 0.005% to about 1%, about 0.005% to about 0.5%, about 0.005% to about 0.4%, about 0.005% to about 0.3%, about 0.005% to about 0.2%, about 0.005% to about 0.1%, about 0.005% to about 0.05%, about 0.005% to about 0.02%, about 0.005% to about 0.01%, about 0.01% to about 5%, about 0.01% to about 4%, about 0.01% to about 3%, about 0.01% to about 2%, about 0.01% to about 1 %, about 0.01% to about 0.5%, about 0.01% to about 0.4%, about 0.01% to about 0.3%, about 0.01% to about 0.2%, about 0.01% to about 0.1%, about 0.01% to about 0.05%, about 0.01% to about 0.02%, about 0.02% to about 5%, about 0.02% to about 4%, about 0.02% to about 3%, about 0.02% to about 2%, about 0.02% to about 1%, about 0.02% to about 0.5%, about 0.02% to about 0.4%, about 0.02% to about 0.3%, about 0.02% to about 0.2%, about 0.02% to about 0.1%, about 0.02% to about 0.05%, about 0.05% to about 5%, about 0.05% to about 4% , about 0.05% to about 3%, about 0.05% to about 2%, about 0.05% to about 1%, about 0.05% to about 0.5%, about 0.05% to about 0.4%, about 0.05% to about 0.3%, about 0.05% to about 0.2%, about 0.05% to about 0.1%, about 0.1% to about 5%, about 0.1% to about 4%, about 0.1% to about 3%, about 0.1% to about 2%, about 0.1% to about 1%, about 0.1% to about 0.5%, about 0.1% to about 0.4%, about 0.1% to about 0.3%, about 0.1% to about 0.2%, about 0.2% to about 5%, about 0.2% to about 4%, about 0.2% to about 3%, about 0.2% to about 2%, about 0.12% to about 1%, about 0.2% to about 0.5%, about 0.2% to about 0.4%, about 0.2% to about 0.3%, about 0.3% to about 5%, about 0.3% to about 4%, about 0.3% to about 3%, about 0.3% to about 2%, about 0.3% to about 1%, about 0.3% to about 0.5%, about 0.3% to about 0.4%, about 0.4% to about 5%, about 0.4% to about 4%, about 0.4% to about 3%, about 0.4% to about 2%, about 0.3% to about 1%. In some embodiments, the copolymer concentration is from about 0.0001% to about 5% by weight of the composition. In some embodiments, the copolymer concentration is from about 0.001% to about 1%, from about 0.005% to about 0.5%, from about 0.005% to about 0.02%, from about 0.01% to about 0.2%, from about 0.1% to about 0.4%, from about 0.4% to about 1%, from about 0.4% to about 0.5%, from about 0.5% to about 5%, from about 0.5% to about 4%, from about 0.5% to about 3%, from about 0.5% to about 2%, from about 0.5% to about 1%, from about 1% to about 5%, from about 1% to about 4%, from about 1% to about 3%, from about 1% to about 2%, from about 2% to about 5%, from about 2% to about 4%, from about 2% to about 3%, from about 3% to about 5%, from about 3% to about 4%, or from about 4% to about 5%. In some embodiments, the copolymer concentration is from about 0.0001% to about 5% by weight of the composition. In some embodiments, the copolymer concentration is from about 0.001% to about 1%, from about 0.005% to about 0.5%, from about 0.005% to about 0.02%, from about 0.01% to about 0.2%, from about 0.1% to about 0.4%, or from about 0.2% to about 0.3% by weight of the composition. 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. .

[0065] In some embodiments, the protein is selected from antibodies and fragments thereof, cytokines, chemokines, hormones, vaccine antigens, cancer antigens, adjuvants, and combinations thereof. In some embodiments, the compositions of the present invention exhibit reduced protein aggregation compared to compositions of the same protein without the copolymer. In some embodiments, the compositions of the present invention exhibit reduced protein precipitation compared to compositions of the same protein without the copolymer. In some embodiments, the composition comprises a protein concentration that is at least twice, at least three times, at least four times, or at least five times the concentration of the same protein in a composition without the copolymer.

[0066] In some embodiments, the composition comprising the copolymer of the present disclosure and biomolecules also comprises one or more pharmaceutically acceptable carriers, aqueous buffers, tension regulators and preservatives. As used in the present application, pharmaceutically acceptable carriers, tension regulators and preservatives are non-toxic to the recipient at the dosage and concentration adopted. In some embodiments, the composition contains a buffer, such as phosphate, citrate, succinate, other organic acids and histidine, wherein the term "buffer" refers to a mixture of a weak acid and its conjugate base, and vice versa, which is used to maintain the pH of the solution at an almost constant value. In some embodiments, the buffer comprises one or more phosphates. In some embodiments, the buffer is sodium phosphate.

[0067] In some embodiments, the composition contains a tonicity adjuster, such as sodium chloride, potassium chloride, mannitol, glucose, glycerol, or magnesium chloride. In some embodiments, the tonicity adjuster is sodium chloride (NaCl) or glycerol. In some embodiments, the tonicity adjuster is glycerol.

[0068] In some embodiments, the compositions contain one or more preservatives.

[0069] 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 consists essentially of water.

[0070] In some embodiments, the pH of the composition is from about 4 to about 9, such as 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.

[0071] Also provided are compositions comprising the polyacrylamide-based copolymers of the present disclosure and a lipid-based carrier. In some embodiments, the lipid-based carrier is selected from liposomes, polymersomes, micelles, and lipid nanoparticles.

[0072] In some embodiments, the copolymers of the present disclosure are used as formulation additives in formulations comprising one or more biomolecules (eg, biomolecules as described in the present application).

[0073] 3.1.3. Properties of copolymer compositions

[0074] The polyacrylamide-based copolymers of the present disclosure have unique properties and, when used with biomolecules in aqueous formulations, can impart beneficial properties to the formulations.

[0075] In some embodiments, the copolymers described in the present disclosure reduce or prevent aggregation of biomolecules or lipid-based carriers, for example, when formulated in an aqueous formulation containing the biomolecule or lipid-based carrier. In some embodiments, the biomolecule is a protein. Thus, in some embodiments, the copolymers described in the present disclosure reduce or prevent aggregation of proteins, for example, when formulated in an aqueous formulation containing the protein.

[0076] The copolymers can be used with any protein that is prone to aggregation in aqueous media. Non-limiting examples include antibodies and fragments thereof, cytokines, chemokines, hormones, vaccine antigens, cancer antigens, adjuvants, and combinations thereof. In some embodiments, the polyacrylamide-based copolymers reduce or prevent aggregation of proteins that are prone to aggregation in aqueous media.

[0077] The copolymers of the present disclosure can also increase the stability of the preparation. According to the embodiments of the present disclosure, the copolymers described in the present application can be added to improve the stability of the preparation, which contains, for example, biomolecules or lipid-based carriers, such as peptides, proteins and conjugates thereof, nucleic acids and oligonucleotides, liposomes, polymer bodies, micelles and lipid nanoparticles. In some embodiments, the addition of copolymers of the present disclosure increases the stability of protein preparations. In some embodiments, the addition of copolymers of the present disclosure increases the stability of liposome preparations. In some embodiments, the addition of copolymers of the present disclosure increases the stability of micelle preparations. In some embodiments, the addition of copolymers of the present disclosure increases the stability of lipid nanoparticle preparations. In some embodiments, the addition of copolymers of the present disclosure increases the stability of preparations comprising one or more nucleic acids. In some embodiments, the addition of copolymers of the present disclosure increases the stability of preparations comprising one or more messenger RNAs (mRNAs). In some embodiments, the addition of copolymers of the present disclosure increases the stability of preparations comprising one or more small interfering RNAs (siRNAs). In some embodiments, the addition of copolymers of the present disclosure increases the stability of preparations comprising one or more deoxyribonucleic acids (DNAs).

[0078] In some embodiments, copolymers of the present disclosure improve the thermal stability of preparations containing biomolecules, in some embodiments, the biomolecules are proteins. Protein preparations generally require expensive refrigerated transportation and storage to prevent the loss of protein integrity. Maintaining protein integrity is a challenge for the pharmaceutical industry, health care providers, and patients worldwide, particularly in developing and low-income areas where the cold chain required to maintain protein efficacy and effectiveness is imperfect, overburdened, or nonexistent. This results in a large amount of therapeutic protein preparations being wasted and potentially endangering the lives of patients. Therefore, cold chain interruptions may be costly. Copolymers of the present disclosure can impart long-term stability and / or cold chain flexibility to protein preparations. For example, although commercial protein preparations have a good shelf life when properly stored (i.e., refrigerated), cold chain interruptions can reduce the biological activity and preparation integrity of the protein.

[0079] In some embodiments, protein formulations containing copolymers of the present invention are stable for a long time at a temperature higher than that usually required for cold chain storage. For example, protein formulations containing copolymers of the present invention can be stored at room temperature (between about 20°C and about 22°C) or at elevated temperatures, such as about 25°C, about 30°C, about 35°C, about 40°C, about 45°C or higher temperatures, while maintaining protein integrity. In some embodiments, protein formulations containing copolymers of the present invention can be stored at a temperature higher than that usually 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 copolymer enables protein formulations to be stored at room temperature or higher temperatures without the need for cold chain storage. In some embodiments, cold chain storage is not required to maintain protein integrity.

[0080] In some embodiments, the use of the copolymers of the present disclosure in protein formulations maintains formulation integrity, bioactivity, pharmacokinetics, and pharmacodynamics over longer periods of time when exposed to conditions such as elevated temperature and agitation compared to the same formulation without the copolymers. In some embodiments, the formulations of the present invention can maintain protein integrity at elevated temperatures 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 compared to the same formulation without the copolymers of the present disclosure.

[0081] In some embodiments, copolymers of the present disclosure reduce the aggregation rate of biomolecules in aqueous formulations. In some embodiments, 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, a cytokine, a chemokine, a hormone, a vaccine antigen, a cancer antigen, an adjuvant, and a combination thereof. In some embodiments, this allows the preparation of a more concentrated protein solution, thereby reducing the volume of the protein solution that needs to be administered to, for example, a patient. In some embodiments, the concentration of the protein included in the composition is at least twice, at least three times, at least four times, or at least five times the concentration of the same protein in the composition without the copolymer.

[0082] 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 protein in a composition comprising the copolymers of the present disclosure exhibits increased stability when stored at room temperature compared to a protein composition without the copolymers. In some embodiments, the increased stability is at least 10 times, at least 15 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, at least 50 times, or more, compared to a protein composition without the copolymers.

[0083] In some embodiments, the copolymers of the present disclosure maintain protein activity during 6 months of stress aging without changing one or more of the formulation pharmacokinetics, protein secondary structure, formulation transparency, and in vivo biological activity. In some embodiments, the copolymers of the present invention reduce protein aggregation in the formulation when stored at ambient temperature (about 23°C to about 27°C) compared to protein formulations without the copolymers. In some embodiments, the copolymers of the present invention reduce protein aggregation in the formulation when stored at 37°C compared to protein formulations without the copolymers. In some embodiments, the copolymers of the present invention reduce protein aggregation in the formulation when stored at 50°C compared to protein formulations without the copolymers.

[0084] In some embodiments, the copolymer of the present invention increases the time to aggregation of the protein formulation at least 10 times, at least 15 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, or at least 50 times when stored at 37°C, compared to the protein formulation without the copolymer. The time to aggregation can be assessed by any known method, including, for example, by transmittance measurement. In some embodiments, the copolymer of the present invention increases the time to aggregation of the protein formulation at least 10 times when stored at 37°C, compared to the protein formulation without the copolymer. In some embodiments, the copolymer of the present invention increases the time to aggregation of the protein formulation at least 20 times when stored at 37°C, compared to the protein formulation without the copolymer. In some embodiments, the copolymer of the present invention increases the time to aggregation of the protein formulation at least 30 times when stored at 37°C, compared to the protein formulation without the copolymer. In some embodiments, the copolymer of the present invention increases the time to aggregation of the protein formulation at least 40 times when stored at 37°C, compared to the protein formulation without the copolymer. In some embodiments, the copolymers of the present invention increase the time to aggregation of a protein formulation at least 50 times when stored at 37°C compared to a protein formulation without the copolymer. In some embodiments, the copolymers of the present invention increase the time to aggregation of a protein formulation at least 10 times, at least 15 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, or at least 50 times when stored at 50°C compared to a protein formulation without the copolymer. In some embodiments, the copolymers of the present invention increase the time to aggregation of a protein formulation at least 10 times when stored at 50°C compared to a protein formulation without the copolymer. In some embodiments, the copolymers of the present invention increase the time to aggregation of a protein formulation at least 20 times when stored at 50°C compared to a protein formulation without the copolymer. In some embodiments, the copolymers of the present invention increase the time to aggregation of a protein formulation at least 30 times when stored at 50°C compared to a protein formulation without the copolymer. In some embodiments, the copolymers of the invention increase the time to aggregation of a protein formulation at least 40-fold when stored at 50° C. compared to a protein formulation without the copolymer. In some embodiments, the copolymers of the invention increase the time to aggregation of a protein formulation at least 50-fold when stored at 50° C. compared to a protein formulation without the copolymer.

[0085] In addition to refrigerated transportation, in the early stages of the cold chain, maintaining appropriate transportation and storage conditions during local distribution and when reaching the patient's hands poses a challenge in many parts of the world. As described in the present application, the addition of a polyacrylamide-based copolymer can maintain the integrity of the protein formulation even during severe cold chain interruptions. In some embodiments, this reduces the cold chain requirements for protein transportation and storage that are difficult to maintain in resource-poor environments. As disclosed in the present application, polyacrylamide-based copolymers can improve cold chain resilience as formulation additives, thereby expanding global access to key 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.

[0086] In some embodiments, the presence of the copolymer in a formulation containing a protein does not alter the pharmacokinetic (PK) or pharmacodynamic (PD) properties of the active ingredient within the formulation.

[0087] 3.2. Usage

[0088] As summarized above, provided in the present disclosure are methods of using the compositions and combined formulations described in the present disclosure, which include polyacrylamide-based copolymers containing a water-soluble carrier monomer and a functional dopant monomer (eg, DEA).

[0089] The present invention provides a method for improving the stability of a formulation containing a biomolecule or a lipid-based carrier. According to embodiments of the present disclosure, the copolymers described in the present application can be added to improve the stability of formulations containing peptides, proteins and conjugates thereof, nucleic acids and oligonucleotides, liposomes, polymer bodies, micelles and lipid nanoparticles. In some embodiments, a method for improving the stability of a protein formulation is provided. In some embodiments, a method for improving the thermal stability of a formulation containing a biomolecule or a lipid-based carrier is provided. In some embodiments, a method for improving the thermal stability of a protein formulation is provided. A method for reducing the aggregation rate of a biomolecule or a lipid-based carrier in an aqueous formulation is also provided. 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, a cytokine, a chemokine, a hormone, a vaccine antigen, a cancer antigen, an adjuvant and a combination thereof.

[0090] In some embodiments, the method comprises adding 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 carrier, for example, adding about 0.001 wt % to about 4 wt %, about 0.001 wt % to about 3 wt %, about 0.001 wt % to about 2 wt %, about 0.001 wt % to about 1 wt %, about 0.001 wt % to about 0.5 wt %, about 0.001 wt % to about 0.4 wt %, about 0.001 wt % to about 0.3 wt %, about 0.001 wt % to about 0.2 wt %, about 0.001 wt % to about 0.1 wt %, about 0.001 wt % to about 0.05 wt %, about 0.001 wt % to about 0.5 wt %, about 0.001 wt % to about 0.6 wt %, about 0.001 wt % to about 0.7 wt %, about 0.001 wt % to about 0.8 wt %. % by weight, about 0.005 wt % to about 0.5 wt %, about 0.005 wt % to about 0.6 wt %, about 0.005 wt % to about 0.8 wt %, about 0.01 wt % to about 0.02 wt %, about 0.001 wt % to about 0.01 wt %, about 0.001 wt % to about 0.005 wt %, about 0.005 wt % to about 5 wt %, about 0.005 wt % to about 4 wt %, about 0.005 wt % to about 3 wt %, about 0.005 wt % to about 2 wt %, about 0.005 wt % to about 1 wt %, about 0.005 wt % to about 0.5 wt %, about 0.005 wt % to about 0.4 wt %, about 0.005 wt % to about 0.3 wt %, about 0.005 wt % to about 0.2 wt %, about 0.005 wt % to about 0.1 wt %, about 0.005 wt % to about 0.05 wt %, about 0.005 wt % to about % to about 0.02% by weight, about 0.005% by weight to about 0.01% by weight, about 0.01% by weight to about 5% by weight, about 0.01% by weight to about 4% by weight, about 0.01% by weight to about 3% by weight, about 0.01% by weight to about 2% by weight, about 0.01% by weight to about 1% by weight, about 0.01% by weight to about 0.5% by weight, about 0.01% by weight to about 0.4% by weight, about 0.01% by weight to about 0.3% by weight, about 0.01% by weight to about 0.2% by weight, about 0.01% by weight to about 0.1% by weight, about 0.01% by weight to about 0.05% by weight, about 0.01% by weight to about 0.02% by weight, about 0.02% by weight to about 5% by weight, about 0.02% by weight to about to about 4 wt%, about 0.02 wt% to about 3 wt%, about 0.02 wt% to about 2 wt%, about 0.02 wt% to about 1 wt%, about 0.02 wt% to about 0.5 wt%, about 0.02 wt% to about 0.4 wt%, about 0.02 wt% to about 0.3 wt%, about 0.02 wt% to about 0.2 wt%, about 0.02 wt% to about 0.1 wt%, about 0.02 wt% to about 0.05 wt%, about 0.05 wt% to about 5 wt%, about 0.05 wt% to about 4 wt%, about 0.05 wt% to about 3 wt%, about 0.05 wt% to about 2 wt%, about 0.05 wt% to about 1 wt%, about 0.05 ...0.05 wt% to about 0.4 wt%, about 0.05 wt% to about 0.3 wt%, about 0.05 wt% to about 0.2 wt%, about 0.05 wt% to about 0.1 wt%, about 0.1 wt% to about 5 wt%, about 0.1 wt% to about 4 wt%, about 0.1 wt% to about 3 wt%, about 0.1 wt% to about 2 wt%, about 0.1 wt% to about 1 wt%, about 0.1 wt% to about 0.5 wt%, about 0.1 wt% to about 0.4 wt%, about 0.1 wt% to about 0.3 wt%, about 0. .1 wt % to about 0.2 wt %, about 0.2 wt % to about 5 wt %, about 0.2 wt % to about 4 wt %, about 0.2 wt % to about 3 wt %, about 0.2 wt % to about 2 wt %, about 0.2 wt % to about 1 wt %, about 0.2 wt % to about 0.5 wt %, about 0.2 wt % to about 0.4 wt %, about 0.2 wt % to about 0.3 wt %, about 0.3 wt % to about 5 wt %, about 0.3 wt % to about 4 wt %, about 0.3 wt % to about 3 wt %, about 0.2 wt % to about 2 wt %. %, about 0.3 wt % to about 1 wt %, about 0.3 wt % to about 0.5 wt %, about 0.3 wt % to about 0.4 wt %, about 0.4 wt % to about 5 wt %, about 0.4 wt % to about 4 wt %, about 0.4 wt % to about 3 wt %, about 0.4 wt % to about 2 wt %, about 0.4 wt % to about 1 wt %, about 0.4 wt % to about 0.5 wt %, about 0.5 wt % to about 5 wt %, about 0.5 wt % to about 4 wt %, about 0.5 wt % to about 3 wt %, about 0.4 wt % to about 2 weight %, about 0.5 weight % to about 1 weight %, about 1 weight % to about 5 weight %, about 1 weight % to about 4 weight %, about 1 weight % to about 3 weight %, about 1 weight % to about 2 weight %, about 2 weight % to about 5 weight %, about 2 weight % to about 4 weight %, about 2 weight % to about 3 weight %, about 3 weight % to about 5 weight %, about 3 weight % to about 4 weight %, about 4 weight % to about 5 weight % or about 0.005 weight %, about 0.01 weight % or about 0.1 weight % of the copolymer based on polyacrylamide of the present disclosure is added to the preparation. In some embodiments, the method includes adding about 0.0005 weight % to about 5 weight % of the copolymer based on polyacrylamide of the present disclosure to the preparation. In some embodiments, the method includes adding about 0.001 weight % to about 1 weight % of the copolymer based on polyacrylamide of the present disclosure to the preparation. In some embodiments, the preparation comprises biomolecules. In some embodiments, the preparation comprises proteins. In some embodiments, the preparation comprises lipid-based carriers. In some embodiments, the lipid-based carrier comprises a liposome, a lipid nanoparticle, a polymersome, or a micelle.

[0091] 3.2.1. Other uses of polyacrylamide-based copolymers

[0092] The polyacrylamide-based copolymers disclosed herein can be used in any composition or formulation in which a surfactant is used. For example, the copolymers disclosed herein can be used in applications including, but not limited to, beauty products, hair products, lotions, food products, nutritional products, pigments, and inks. In some embodiments, the copolymers are used to improve food texture or improve mouthfeel. In some embodiments, the copolymers are used in biopharmaceutical compositions for animal or veterinary use. In some embodiments, the presence of the copolymers improves the stability of the formulation.

[0093] 3.3. Definition

[0094] Unless otherwise defined, all technical and scientific terms used in this application have the same meanings as those generally understood by those of ordinary skill in the art to which the present disclosure belongs. Methods and materials for use in this application are described in this application; other suitable methods and materials known in the art are also used in certain aspects of the present disclosure. Materials, methods and examples are illustrative only and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries and other references mentioned in this application are incorporated by reference in their entirety. In the event of a conflict, this specification (including definitions) shall prevail. When a trade name is used in this application, unless the context otherwise indicates, the trade name includes product preparations, generic drugs and active pharmaceutical ingredients of the trade name product.

[0095] The terms "subject" and "patient" are used interchangeably. The subject can be a mammal, such as a non-primate (e.g., cattle, pigs, horses, cats, dogs, goats, rabbits, rats, mice, etc.) or a primate (e.g., monkeys, apes and humans), such as humans. In certain embodiments, the subject is a mammal, such as a human, diagnosed with a disease or condition provided in the application. In another embodiment, the subject is a mammal, such as a human, at risk of a disease or condition provided in the application. In a specific embodiment, the subject is a human.

[0096] The terms "therapy" and "treatment" are used in the broadest sense as understood in the clinical arts.

[0097] In this disclosure, unless the context clearly dictates otherwise, the terms "a", "an", and "the" are used to include one or more. Unless otherwise specified, the term "or" is used to refer to a non-exclusive "or". The expression "at least one of A and B" has the same meaning as "A, B, or A and B". In addition, it should be understood that the expressions or terms used in this disclosure, unless otherwise defined, are used for descriptive purposes only and not for limiting purposes. The use of any section headings is intended to assist in reading the document and should not be construed as limiting; information related to the section heading may appear within or outside that particular section.

[0098] Values ​​expressed in range format should be interpreted in a flexible manner to include not only the values ​​explicitly listed as the limits of the range, but also all individual values ​​or subranges contained in the range, as if each value and subrange 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 individual values ​​(e.g., 1%, 2%, 3%, and 4%) and subranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the specified range. Unless otherwise specified, the expression "about X to Y" has the same meaning as "about X to about Y". Similarly, unless otherwise specified, the expression "about X, Y, or about Z" has the same meaning as "about X, about Y, or about Z".

[0099] The term "about" as used in this disclosure may allow a degree of variation in a value or range that is within 5% of the stated value or limit of the stated range.

[0100] In the methods described in this disclosure, the actions may be performed in any order unless a time or order of operations is explicitly stated. In addition, specified actions may be performed simultaneously unless explicit claim language states that they are performed separately. For example, a claimed action to perform X and a claimed action to perform Y may be performed simultaneously in a single operation, and the resulting method will fall within the literal scope of the claimed method.

[0101] The term "polymer" refers to a substance or material composed of repeating monomer subunits.

[0102] "Acrylamide monomer" used in this application refers to a monomer type with 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).

[0103] The term "polyacrylamide-based copolymer" refers to a polymer formed by the polymerization of two or more monomer species, wherein at least one of the monomer species has an acrylamide functional group (acrylamide monomer), and the monomers are structurally different. In some embodiments, the polyacrylamide-based copolymer is formed by the polymerization of two structurally different acrylamide monomers (two structurally different monomers, each of which has an acrylamide functional group). The resulting copolymer can be an alternating copolymer, in which the monomer species are connected in an alternating manner; a random copolymer, in which the monomer species are interconnected within the polymer chain without a definite pattern; a block copolymer, in which a polymer block of one monomer species is connected to a polymer block composed of another monomer species; and a graft copolymer, in which the main polymer chain consists of one monomer species, and a polymer block of another monomer species is connected to the main polymer chain as a side branch. In some embodiments, the polyacrylamide-based copolymer of the present disclosure is formed by the polymerization of a water-soluble carrier monomer and a functional dopant monomer. In some embodiments, the polyacrylamide-based copolymer of the present disclosure is a random copolymer.

[0104] As defined in this application, the term "water-soluble carrier monomer" refers to acrylamide monomer species, which is a water-soluble species within the 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 a shell-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 an interface between a liquid and a tube. In some embodiments, the interface is an interface between a liquid and a catheter. In some embodiments, the shell-water interface is located in a pump system. In some embodiments, the shell-water interface is in a closed loop system. In some embodiments, the water-soluble carrier monomer is non-ionic. 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).

[0105] The term "functional dopant monomer" used in this application refers to a type of acrylamide monomer having physicochemical properties (e.g., hydrophobicity, charge) different from water-soluble carrier monomers. In some embodiments, the functional dopant monomers in the copolymer based on polyacrylamide promote the binding of polymers to interfaces; such interfaces may 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. Functional dopant monomers can act as stabilizing parts to promote interactions with biomolecules (e.g., proteins, peptides, antibodies, antibody-drug conjugates, nucleic acids, lipid particles, and combinations thereof) (e.g., to prevent biomolecule aggregation). Functional dopant monomers can be further divided into hydrogen bonding, ionic, hydrophobic, and aromatic monomers according to their chemical composition. Typically, functional dopant monomers are copolymerized at a lower weight percentage than the water-soluble carrier monomers.

[0106] The term "polymerization" refers to the process by which monomer molecules chemically react to form polymer chains or three-dimensional networks. 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.

[0107] The term "degree of polymerization" (DP) refers to the number of monomer units in a polymer. It is calculated by dividing the mean molecular weight of a polymer sample by the molecular weight of a monomer. As defined in the present application, the mean molecular weight of a polymer can be represented by number average molecular weight (Mn), weight average molecular weight (Mw), Z average molecular weight (Mz) or the molecular weight (Mp) at the peak maximum of a molecular weight distribution curve. The mean molecular weight of a polymer can be determined by a variety of 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 (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 suitable analytical methods known in the art, such as nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy (FT-IR) and Raman spectroscopy.

[0108] The term "amphiphilic" refers to a chemical substance that has both hydrophilicity (water-loving, polar) and lipophilicity (fat-loving, non-polar). Examples of common amphiphilic compounds include detergents, soaps, surfactants, lipoproteins, and phospholipids. In some embodiments, the amphiphilic substance is a charged substance. In some embodiments, the amphiphilic substance is a neutral substance.

[0109] "Lipid-based carrier" as used in this application refers to a structure with a protective outer layer of lipids that can be used as a drug delivery vehicle. For example, lipid-based carriers can be used to encapsulate and transport cargo (e.g., therapeutic agents) to biological targets. Examples of lipid-based carriers include, but are not limited to, liposomes, micelles, polymer bodies, and lipid nanoparticles.

[0110] As used in this application, "biomolecules" refer to molecules such as proteins, nucleic acids, polysaccharides and lipids.

[0111] The term "protein" is defined as a class of macromolecules comprising one or more long chains of amino acids. A wide variety of proteins can be considered to belong to a protein family based on having similar structural features, having specific biological functions and / or being associated with specific microorganisms (particularly pathogenic microorganisms). Such proteins include, for example, antibodies (immunoglobulins), cytokines, chemokines, enzymes, hormones, vaccine antigens, cancer antigens, adjuvants, nutritional markers, and tissue-specific antigens.

[0112] The term "nucleic acid" as used in this application includes deoxyribonucleic acid (DNA), ribonucleic acid (RNA), messenger RNA (mRNA), small interfering RNA (siRNA), short hairpin RNA (shRNA) and micro RNA (miRNA).

[0113] The term "antibody" refers to a large Y-shaped protein produced by the immune system to identify and neutralize foreign objects such as pathogenic bacteria and viruses. The term "antibody" includes monoclonal antibodies (e.g., full-length or complete monoclonal antibodies), polyclonal antibodies, multivalent antibodies, multispecific antibodies (e.g., bispecific or trispecific antibodies, as long as they show the desired biological activity), and may also include certain antibody fragments. The antibody may be a human antibody, a humanized antibody, and / or an affinity matured antibody."Antibody fragment" only comprises a portion of a complete antibody, wherein in certain embodiments, the portion retains at least one of the functions associated with the portion when present in the complete antibody, typically most or all. In one embodiment, the antibody fragment comprises the antigen binding site of the complete antibody, thus retaining the ability to bind antigens. In another embodiment, the antibody fragment (e.g., an antibody fragment comprising an Fc region) retains at least one of the biological functions associated with the Fc region when in the complete antibody, such as FcRn combination, antibody half-life regulation, ADCC function, and complement binding. In one embodiment, the antibody fragment is a monovalent antibody, and its in vivo half-life is substantially similar to that of the complete antibody. For example, such an antibody fragment may comprise an antigen binding arm linked to an Fc sequence that confers in vivo stability to the fragment.

[0114] The term "aggregation" refers to the formation of higher molecular weight amorphous species due to the non-covalent adhesion ("clumping") of smaller species. Aggregation processes can be irreversible or reversible. Aggregation can occur in many biological and synthetic molecules, including proteins, peptides, lipid particles, nucleic acids, inorganic nanoparticles and organic nanoparticles (e.g., micelles, lipid nanoparticles, liposomes, polymersomes), which may also include encapsulated substances.

[0115] In the case of protein aggregation, the formation of protein aggregates can be due to the disordered nature of the protein itself, or the misfolding of protein molecules, which results in the exposure of hydrophobic residues and surfaces that are usually buried inside the three-dimensional structure of the protein. Due to the hydrophobic effect, the exposed hydrophobic parts of the misfolded proteins tend to interact with other misfolded protein molecules to shield the exposed hydrophobic surfaces, which can lead to protein aggregation.

[0116] Some biomolecules are more "aggregate-prone" than others. For example, the amino acid sequence and overall three-dimensional structure of a protein are related to its aggregation sensitivity. For example, transmembrane proteins are more (or more) aggregated than non-membrane proteins, especially when recombinantly expressed without the use of stabilizers. Proteins subjected to conditions other than physiological conditions (37°C, -neutral pH, isotonic) may also be more likely to aggregate than in their natural 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 result in the formation of microscopic or visible particles (i.e., precipitates). The extent of protein aggregation visible under a microscope can be measured by a variety of analytical methods known in the art, such as size exclusion chromatography (SEC), gel electrophoresis, asymmetric field flow separation (AF4), analytical ultracentrifugation (AUC), mass spectrometry (MS), optical microscopy, fluorescence microscopy, dynamic light scattering (DLS), multi-angle laser light scattering (MALLS), microfluidic imaging, turbidity / turbidimetry, and transmittance measurement.

[0117] As used in this application, the term "reduced aggregation" of a biomolecule or lipid-based carrier includes all forms of reduced aggregation. Compared to the same biomolecule or lipid-based carrier composition without the polyacrylamide-based copolymer of the present invention, the observed degree of aggregation or the amount of aggregation (e.g., in the composition) can be reduced. Therefore, "reduced aggregation" includes no observable aggregation or a reduction in the amount of aggregation (e.g., a reduction in the level of aggregated protein). Therefore, the amount of aggregates present in the composition of the present invention can be reduced by 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% compared to the amount of aggregates of the same biomolecule or lipid-based carrier without 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 field flow fractionation (AF4), analytical ultracentrifugation (AUC), mass spectrometry (MS), optical microscopy, fluorescence microscopy, dynamic light scattering (DLS), multi-angle laser light scattering (MALLS), microfluidic imaging, turbidity / nephelometric assays, and transmittance measurements.

[0118] As used in this application, the term "increased stability", when referring to a formulation containing a biomolecule or a lipid-based carrier, refers to a measurable reduction in the amount of aggregation over a fixed period of time under test or fixed storage conditions compared to the amount of aggregation of the same biomolecule or lipid-based carrier without the polyacrylamide-based copolymer.

[0119] As used in this application, the term "aggregated protein" or "protein aggregate" refers to a collection of proteins that are disordered or misfolded and clumped together. Aggregates can be soluble or insoluble. Protein aggregates include, but are not limited to, inclusion bodies, soluble and insoluble precipitates, soluble non-natural oligomers, gels, fibrils, film bodies, filaments, protofibrils, amyloid deposits, amyloid protofibrils, plaques, and dispersed non-natural intracellular oligomers. In some embodiments, the proteins in the protein aggregates are soluble precursors before they aggregate. Protein aggregation can be prevented in a composition containing a polyacrylamide-based copolymer of the present invention. Protein aggregation can also be reduced in a composition containing a polyacrylamide-based copolymer of the present invention compared to a composition containing the same protein but without the polyacrylamide-based copolymer of the present invention. Therefore, the polyacrylamide-based copolymer can reduce or prevent the aggregation of proteins. 4. Other Implementations

[0120] The present disclosure is further described by the following non-limiting terms:

[0121] Item 1. A polyacrylamide-based copolymer comprising:

[0122] a water-soluble carrier monomer selected from the group consisting of N-(3-methoxypropyl)acrylamide (MPAM), 4-acryloylmorpholine (MORPH), N,N-dimethylacrylamide (DMA), N-hydroxyethylacrylamide (HEAM), and acrylamide (AM), and combinations thereof; and

[0123] A functional dopant monomer comprises N,N-diethylacrylamide (DEA).

[0124] Item 2. The copolymer according to Item 1, wherein: the water-soluble carrier monomer is selected from N-(3-methoxypropyl)acrylamide (MPAM) and 4-acryloylmorpholine (MORPH).

[0125] Item 3. The copolymer according to Item 1 or Item 2, wherein the weight percentage (wt %) of the water-soluble carrier monomer is 70% to 98%.

[0126] Item 4. The copolymer according to any one of Items 1 to 3, wherein the weight percentage (wt %) of N,N-diethylacrylamide (DEA) is 2% to 30%.

[0127] Item 5. The copolymer according to Item 4, wherein the weight percentage (wt %) of N,N-diethylacrylamide (DEA) is 2% to 16%.

[0128] Item 6. The copolymer according to any one of Items 2-5, wherein the water-soluble carrier monomer is MORPH.

[0129] Item 7. The copolymer according to any one of Items 2-5, wherein the water-soluble carrier monomer is MPAM.

[0130] Item 8. A copolymer according to any one of Items 1 to 7, wherein the copolymer further comprises 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) and combinations thereof.

[0131] Item 9. The copolymer according to any one of Items 1 to 8, wherein the degree of polymerization is 10 to 500.

[0132] Item 10. The copolymer according to Item 9, wherein the degree of polymerization is 20 to 200.

[0133] Item 11. The copolymer according to Item 10, wherein the degree of polymerization is 50.

[0134] Item 12. The copolymer according to any one of Items 1 to 11, wherein the molecular weight of the copolymer is 1,000 to 40,000 g / mol.

[0135] Item 13. The copolymer according to Item 12, wherein the copolymer has a molecular weight of 2,000 to 10,000 g / mol.

[0136] Item 14. The copolymer according to Item 13, wherein the copolymer has a molecular weight of 4,000 to 6,000 g / mol.

[0137] Item 15. The copolymer according to any one of Items 1 to 14, wherein the copolymer is amphiphilic.

[0138] Item 16. A polyacrylamide-based copolymer comprising:

[0139] a water-soluble carrier monomer comprising an acrylamide-reactive moiety; and

[0140] A functional dopant monomer comprising N,N-diethylacrylamide (DEA);

[0141] in:

[0142] The weight percentage (wt%) of the water-soluble carrier monomer is 70% to 98%;

[0143] The weight percentage (wt%) of the functional dopant monomer is 2% to 30%;

[0144] The polyacrylamide-based copolymer has an average molecular weight (Mn) of about 1,000 g / mol to about 30,000 g / mol; and

[0145] The degree of polymerization is from about 10 to about 250.

[0146] Item 17. The copolymer according to Item 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).

[0147] Item 18. The copolymer according to Item 17, wherein the water-soluble carrier monomer is selected from N-(3-methoxypropyl)acrylamide (MPAM) and 4-acryloylmorpholine (MORPH).

[0148] Item 19. A copolymer according to any one of Items 16-18, wherein the functional dopant monomer further comprises a 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) and combinations thereof.

[0149] Item 20. A composition comprising the copolymer according to any one of Items 1 to 19 and a pharmaceutically acceptable excipient.

[0150] Item 21. A composition comprising the copolymer of any one of Items 1 to 19, wherein the composition is a cosmetic product, a hair product, an emulsion, a food product, a veterinary product, or a nutritional product.

[0151] Item 22. A composition comprising the copolymer of any one of Items 1 to 19 and a protein.

[0152] Item 23. The composition according to Item 22, wherein the protein is a protein that is susceptible to aggregation in aqueous media.

[0153] Item 24. The composition of any one of Items 20-23, wherein the copolymer concentration is 0.1% to about 10% by weight of the composition.

[0154] Item 25. The composition of Item 24, wherein the copolymer concentration is 0.5% or about 5% by weight of the composition.

[0155] Item 26. The composition of Item 25, wherein the copolymer concentration is 1% by weight of the composition.

[0156] Item 27. The composition according to any one of Items 20-26, wherein the composition comprises one or more of an aqueous buffer, a tonicity adjuster, and a preservative.

[0157] Item 28. The composition according to any one of Items 20-27, wherein the composition is an aqueous composition.

[0158] Item 29. The composition according to Item 28, wherein the pH of the composition is 4 to 9.

[0159] Item 30. The composition according to Item 28, wherein the pH of the composition is from about 6 to about 8.

[0160] Item 31. A method for improving the thermal stability of a protein formulation, the method comprising adding 0.0005 wt % to 5 wt % of the copolymer described in any one of Items 1-19 to the protein formulation.

[0161] Item 32. A method for improving the stability of a protein formulation, the method comprising adding about 0.005 wt % to about 5 wt % of the copolymer of any one of Items 1-19 to the protein formulation.

[0162] Item 33. A method for reducing the aggregation rate of a protein in an aqueous composition, the method comprising adding 0.005 wt % to 5 wt % of the copolymer described in any one of Items 1-19 to the protein preparation.

[0163] Item 34. The method according to any one of Items 31-33, wherein the protein is a protein that aggregates in an aqueous medium. 5. Examples

[0164] The examples in this section are provided by way of illustration and not limitation. These examples may represent only some embodiments, and it should be understood that the following examples are only illustrative and not limiting. Unless otherwise stated, all substituents are as defined above. Those of ordinary skill in the art can easily obtain the reagents and starting materials. The specific synthesis steps of each route described can be combined in different ways, or combined with steps from different schemes, to prepare the compounds described in this application.

[0165] 5.1. Example 1 - Synthesis of polyacrylamide-based copolymers

[0166] Synthesis of the first copolymer library by automated parallel synthesis: The copolymerization of the carrier and the dopant was carried out using RAFT polymerization ([total monomers] / [CTA]=50, [CTA] / [AIBN]=0.2). The MPAM, MORPH and DMA carrier monomers copolymerized with the DEA dopant monomer were polymerized in DMF, using 2-CPDT or BM1433 as CTA and AIBN as initiator. The total vinyl monomer molar concentration was maintained at 2.72M (MPAM copolymerization), 2.86M (MORPH copolymerization) and 3.84M (DMA copolymerization), so that the homopolymerization of the carrier monomer in DMF will be carried out at a constant 40wt.%. The HEAM carrier monomer copolymerized with the DEA dopant monomer was polymerized in a DMF / EtOH mixture, using 2-CPDT or BM1433 as CTA and AIBN as initiator. The total vinyl monomer molar concentration was maintained at 2.58M (HEAM copolymerization), so that the homopolymerization reaction of HEAM in DMF was carried out at a constant 30wt.%. AM carrier monomer copolymerized with DEA ​​dopant monomer was polymerized in DMF / water mixture using BM1433 as CTA and ACVA as initiator. The total vinyl monomer molar concentration was kept at 4.05 M (AM copolymerization) so that homopolymerization of AM in DMF was carried out at a constant 30 wt.%.

[0167] A reaction mixture was prepared by combining stock solutions of: (i) support, (ii) dopant and (iii) CTA and initiator.

[0168] Polyacrylamide-based copolymer excipients (also known as AC / DC excipients) were synthesized in parallel on a Chemspeed Swing XL automated synthesizer robot equipped with a 4-needle tool and an iSynth reactor. The reaction was carried out in an 8 mL disposable iSynth reactor vial. Aspiration and distribution of all reagent solutions were performed using a 4-needle tool equipped with 2x 10 mL and 2x 1 mL syringes (both 1 mL and 10 mL syringes were equipped with septum puncture needles). All solvent lines were filled with 60 mL (syringe volume of 6 strokes) of degassed DMF. For a 1 mL syringe, the typical aspiration and distribution rate of the reagent was 10 mL / min. During aspiration using the 4-needle tool, a 50 μL air gap and an additional volume of 50 μL were used for the 1 mL syringe, and a 50 μL air gap and an additional volume of 100 μL were used for the 10 mL syringe. After each reagent dispensing task, the needle and tubing were flushed with 3 mL of internal and external volume of starting solvent (for 1 mL syringes) and 20 mL of internal and external volume of starting solvent (for 10 mL syringes). The DMF reservoir was degassed by continuous nitrogen purge. All stock solutions were prepared in reagent bottles with septa and degassed by purging with argon for 15 minutes before being transferred to the Chemspeed. The atmosphere inside the Chemspeed was reduced to <1% oxygen by purging with nitrogen while closing the exhaust port. The reactor vials were exposed to a stream of nitrogen until the reaction started. Calculated aliquots of stock solutions and solvents were transferred to the reactor by an automated liquid handling system. After dispensing, the reactor vials were manually sealed in an inert gas, removed from the Chemspeed, manually shaken to mix the reagents, and heated to 65°C in an oven for 24 hours, after which the reaction vials were cooled to room temperature and exposed to air.

[0169] The procedure for removing CTA Z groups from AC / DC excipients containing MORPH, DMA, HEAM and MPAM copolymers was adapted from the literature. The reaction vial was diluted to 6 mL with DMF. LPO (2 equiv.) and AIBN (20 equiv.) were added to the reaction mixture and sealed with a cap using a PTFE seal. The reaction mixture was purged with nitrogen for 10 minutes while heating at 90°C and then heated at 90°C for 12 hours. The procedure for removing CTA Z groups from AC / DC excipients containing AM copolymers was adapted from the literature. The reaction vial was diluted to 5 mL with miliQ water. H2O2 (20 equiv.) was added to the reaction vial, which was sealed and heated to 60°C for 12 hours. The resulting copolymer was isolated by precipitation as described below.

[0170] AC / DC excipients synthesized with AM and HEAM carriers were precipitated twice from acetone. AC / DC excipients synthesized with DMA and MORPH were precipitated twice from ether. AC / DC excipients synthesized with MPAM were precipitated twice from a mixture of ether and hexane (3:1 ratio). SEC in DMF and poly(ethylene glycol) standards were used to determine the number average molecular weight (Mn) and weight average molecular weight (Mw) and dispersity of AC / DC excipients containing MORPH, MPAM, DMA and HEAM. Aqueous SEC-MALLS was used to determine the Mn, Mw and dispersity of AC / DC excipients containing AM.

[0171] Synthesis of the second polymer library : The typical procedure for the synthesis of MORPH-DEA AC / DC excipient is as follows and is almost identical for all other carrier / dopant combinations, where only the choice and concentration of carrier / dopant are changed. MORPH (41.5 eq.), DEA (8.5 eq.), 2CPDT (1 eq.) and AIBN (0.2 eq.) were mixed in an 8 mL scintillation vial equipped with a PTFE septum and diluted with DMF to a total volume of 2.25 mL (33.3 w / v vinyl monomer concentration). The reaction mixture was purged with nitrogen for 10 minutes and then heated at 65 °C for 12 hours. In order to remove the Z end of the resulting polymer, AIBN (20 eq.) and LPO (2 eq.) were added to the reaction mixture, which was then purged with nitrogen for 10 minutes and heated at 90 °C for 12 hours. The removal of the Z group was confirmed by the ratio of the refractive index to the UV (0-=310 nm) intensity in the SEC analysis. The resulting polymer was precipitated three times from diethyl ether and dried under vacuum overnight. By 1 The resulting composition and molecular weight were determined by HNMR spectroscopy and SEC using poly(ethylene glycol) standards.

[0172] Copolymer Molecular Weight Characterization : Mn, Mw and dispersity of copolymers containing HEAM, DMA, MPAM and MORPH carrier monomers were determined using poly(ethylene glycol) standards (American Polymer Standards) implemented by SEC after passing through two size exclusion chromatography columns.

[0173] Mn, Mw and dispersity of copolymers containing AM were 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 was 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. In this medium, the dn / dc value of the AM copolymer was assumed to be 0.185.

[0174] Method for determining experimental VMwt.% values : Processing of sticky monomers (HEAM, MPAM, MORPH) through Chemspeed can result in monomer loadings that are different from the target monomer loadings selected for copolymerization during the initial AC / DC copolymer library synthesis. The experimental weight percentages are approximated from the peak molecular weight (Mp) of the SEC trace. Since only sticky monomers are affected, changes in Mp are caused by insufficient addition of carrier monomer (in addition to small changes in permeation volume for different weight percentages of dopant monomer compared to carrier). Therefore, to calculate the experimental weight percentages, determine the Mp,max for a given carrier / dopant pair.

[0175] High-throughput synthesis of polyacrylamide libraries : An AC / DC excipient library was combinatorially synthesized by statistical copolymerization of water-soluble carrier monomers and functional dopant DEA monomers. The carrier monomer is the main species responsible for both maintaining solubility and providing an inert barrier to prevent protein-protein interactions. The functional dopant can be copolymerized at a lower weight percentage and statistically incorporated throughout the resulting copolymer. The dopant DEA is selected by design to promote polymer-interface interactions or polymer-protein interactions. The library targets a copolymer with a degree of polymerization (DP) of 50. The experimental degree of polymerization (DP) of the carrier monomer is approximated using equation (SI).

[0176]

[0177] This value is used to approximate the experimental weight percentage (wt%). 1 H NMR and SEC determined the experimental weight percent values ​​of the carrier monomer and the functional dopant monomer for each copolymer.

[0178] The library was generated by parallel synthesis using a Chemspeed Swing XL automated synthesizer (a liquid handling robot in an inert environment). RAFT polymerization was implemented because it provides accurate copolymerization stoichiometry, low dispersity, and controlled molecular weight for a wide range of monomers. Due to the range and availability of commercially available water-soluble monomers (carriers) and functional monomers (dopants) and polymer stability, polyacrylamide derivatives are used for both carrier monomers and dopant monomers. Although monomeric acrylamide derivatives generally exhibit acute toxicity, polyacrylamide derivatives, when properly purified from their monomer precursors, exhibit a high degree of biocompatibility. In addition, the reactivity ratio between various acrylamide monomers is close to 1, and the resulting copolymers have almost no to no dopant gradient composition. Carrier monomers include acrylamide (AM), hydroxyethyl acrylamide (HEAM), dimethyl acrylamide (DMA), acryloyl morpholine (MORPH), and methoxypropyl acrylamide (MPAM) because they are non-ionic and soluble in water (in order of increasing hydrophobicity). The dopant monomer is N,N-diethylacrylamide (DEA), which can be further classified as a hydrophobic monomer based on its chemical composition. 6. Equivalent solutions and citation mergers

[0179] While various embodiments of the present disclosure have been particularly shown and described with reference to a preferred embodiment and various alternative embodiments, those skilled in the relevant art will understand that various changes in form and details may be made therein without departing from the spirit and scope of the present disclosure.

[0180] 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, to the same extent 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 copolymer based on polyacrylamide, comprising: a water-soluble carrier monomer selected from the group consisting of 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 comprises N,N-diethylacrylamide (DEA).

2. The copolymer according to claim 1, wherein: The water-soluble carrier monomer is selected from N-(3-methoxypropyl) acrylamide (MPAM) and 4-acryloylmorpholine (MORPH).

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

4. The copolymer according to any one of claims 1 to 3, wherein the weight percentage (wt%) of N,N-diethylacrylamide (DEA) is 2% to 30%.

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

6. The copolymer according to any one of claims 2 to 5, wherein the water-soluble carrier monomer is MORPH.

7. The copolymer according to any one of claims 2 to 5, wherein the water-soluble carrier monomer is MPAM.

8. The copolymer according to any one of claims 1 to 7, wherein the copolymer further comprises a functional dopant monomer selected from the group consisting of 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) and combinations thereof.

9. The copolymer according to any one of claims 1 to 8, wherein the degree of polymerization is from 10 to 500.

10. The copolymer according to claim 9, wherein the degree of polymerization is 20 to 200. The copolymer according to claim 10 , wherein the degree of polymerization is 50.

12. The copolymer of any one of claims 1-11, wherein the copolymer has a molecular weight of 1,000 to 40,000 g / mol.

13. The copolymer of claim 12, wherein the copolymer has a molecular weight of 2,000 to 10,000 g / mol.

14. The copolymer of claim 13, wherein the copolymer has a molecular weight of 4,000 to 6,000 g / mol.

15. The copolymer according to any one of claims 1 to 14, wherein the copolymer is amphiphilic.

16. A polyacrylamide-based copolymer comprising: a water-soluble carrier monomer comprising an acrylamide-reactive moiety; and A functional dopant monomer comprising N,N-diethylacrylamide (DEA); in: The weight percentage (wt%) of the water-soluble carrier monomer is 70% to 98%; The weight percentage (wt%) of the functional dopant monomer is 2% to 30%; The polyacrylamide-based copolymer has an average molecular weight (Mn) of about 1,000 g / mol to about 30,000 g / mol; and The degree of polymerization is from about 10 to about 250.

17. The copolymer according to claim 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).

18. The copolymer according to claim 17, wherein the water-soluble carrier monomer is selected from N-(3-methoxypropyl)acrylamide (MPAM) and 4-acryloylmorpholine (MORPH).

19. The copolymer according to any one of claims 16-18, wherein the functional dopant monomer further comprises a monomer selected from the group consisting of 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) and combinations thereof.

20. A composition comprising the copolymer according to any one of claims 1 to 19 and a pharmaceutically acceptable excipient.

21. A composition comprising the copolymer according to any one of claims 1 to 19, wherein the composition is a cosmetic product, a hair product, a lotion, a food product, a veterinary product or a nutritional product.

22. A composition comprising the copolymer according to any one of claims 1 to 19 and a protein.

23. The composition of claim 22, wherein the protein is a protein that tends to aggregate in aqueous media.

24. The composition of any one of claims 20-23, wherein the concentration of the copolymer is from 0.1% to about 10% by weight of the composition.

25. The composition of claim 24, wherein the concentration of the copolymer is 0.5% or about 5% by weight of the composition.

26. The composition of claim 25, wherein the concentration of the copolymer is 1% by weight of the composition.

27. The composition of any one of claims 20-26, wherein the composition comprises one or more of an aqueous buffer, a tonicity adjuster, and a preservative.

28. The composition according to any one of claims 20-27, wherein the composition is an aqueous composition.

29. The composition of claim 28, wherein the pH of the composition is 4 to 9.

30. The composition of claim 28, wherein the pH of the composition is from about 6 to about 8.

31. A method for improving the thermal stability of a protein formulation, the method comprising adding 0.0005 wt% to 5 wt% of the copolymer according to any one of claims 1-19 to the protein formulation.

32. A method of improving the stability of a protein formulation, the method comprising adding to the protein formulation from about 0.005 wt % to about 5 wt % of the copolymer of any one of claims 1-19.

33. A method of reducing the rate of protein aggregation in an aqueous composition, the method comprising adding to the protein formulation 0.005% to 5% by weight of the copolymer according to any one of claims 1 to 19.

34. The method of any one of claims 31-33, wherein the protein is a protein that aggregates in an aqueous medium.

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