Injectable formulations comprising a biopharmaceutical agent and a polyacrylamide-based copolymer - Patent Application 20070122997

Abstract

The present disclosure provides an injectable pharmaceutical composition comprising particles suspended in a liquid carrier, the particles comprising a biopharmaceutical agent and a polyacrylamide-based copolymer. The inventors have demonstrated that certain polyacrylamide-based copolymers can be used as stabilizing excipients in particle formulations of biopharmaceutical agents without directly interacting with the biopharmaceutical agent or altering its pharmacokinetic properties. The pharmaceutical composition can be formulated for injection into a patient. Also provided are a syringe loaded with the injectable pharmaceutical composition, a method for administering a therapeutically effective dose of a biopharmaceutical to a subject in need thereof via injection, and a method for preparing the subject injectable pharmaceutical composition.

Description

[Technical Field]

[0001] 1. Background Many biopharmaceuticals in aqueous-based formulations are prone to irreversible aggregation when exposed to high temperatures and / or agitation, necessitating formulation at low concentrations, careful storage, and refrigerated transport (cold chain) to retain activity throughout their shelf life. Maintaining the integrity of various aggregation-prone biopharmaceuticals presents challenges to the global pharmaceutical industry, healthcare providers, and those requiring treatment with such biopharmaceuticals. While the mechanism of aggregation can vary among biopharmaceutical agents, their tendency to aggregate at interfaces increases with formulation concentration, adversely affecting overall formulation stability. Such inherent concentration and / or stability limitations of biopharmaceutical formulations often necessitate the intravenous (IV) transfusion of large volumes of biopharmaceutical therapies at low concentrations. However, IV administration of such formulations places a burden on patients, often requiring lengthy transfusion procedures and access to clinical infrastructure, excluding large at-risk populations from effective treatment. [Background technology]

[0002] Many commercially available excipients are used in an attempt to overcome the challenges associated with formulating biopharmaceutical compounds. These systems can be limited by their critical micelle concentration, potential toxicity due to oxidative degradation, and undesirable interactions between the excipient and the cargo in the bulk.

[0003] Therefore, there is a need for improved injectable biopharmaceutical formulations. Summary of the Invention [Problem to be solved by the invention]

[0004] 2. Overview The present disclosure provides an injectable pharmaceutical composition comprising particles suspended in a liquid carrier. The particles of the composition comprise a biopharmaceutical agent and a polyacrylamide-based copolymer. The inventors have demonstrated that certain polyacrylamide-based copolymers can be used as stabilizing excipients in particulate formulations of biopharmaceutical agents without directly interacting with the biopharmaceutical agent or altering its pharmacokinetic properties. The results presented herein demonstrate that the polyacrylamide-based copolymers of the present disclosure can generally be applied to impart substantial stability benefits to high-concentration particulate compositions of biopharmaceutical agents (such as proteins or peptides) and to modify the injectability and depot formation properties of the resulting compositions.

[0005] In some embodiments, the pharmaceutical composition is formulated for injection into a patient at a biopharmaceutical agent concentration that allows for administration of a therapeutically effective dose in a low-resource environment, as opposed to a composition administered via, for example, IV administration. In some embodiments, the pharmaceutical composition is formulated for administration via subcutaneous injection (SC). In some embodiments, the pharmaceutical composition is formulated for administration via intramuscular injection (IM). In some embodiments, the injectable pharmaceutical composition is shelf-stable.

[0006] According to a first aspect, A particle, Biopharmaceutical agents, and a particle comprising a polyacrylamide-based copolymer; and a liquid carrier in which the particles are suspended.

[0007] Also provided are syringes loaded with the injectable pharmaceutical compositions, methods for administering a therapeutically effective dose of a biopharmaceutical via injection to a subject in need thereof, and methods for preparing the injectable pharmaceutical compositions of the subject. [Brief explanation of the drawings]

[0008] 3. Brief description of the drawings [Figure 1] Panels a-c illustrate the characterization of MoNi. Panel a depicts two size-exclusion chromatography (SEC) traces of MoNi. Panel b shows the H NMR spectrum of MoNi. Panel c depicts the differential scanning calorimetry (DSC) analysis of MoNi at a temperature ramp and cooling rate of 10 °C / min, showing a glass transition temperature of 130-140 °C (top line 1 = cooling, bottom line 2 = heating). [Figure 2] FIG. 1 is a schematic diagram of the spray drying process and formulation of ultra-high concentration (UHC) protein suspensions. [Figure 3] Panels a-c illustrate the injection behavior of exemplary formulations. Panel a illustrates the injection through a 21G needle and the depot formation behavior of Formulation 4 (336 mg / mL BSA in sesame oil with 4-acryloylmorpholine 77%-N-isopropylacrylamide 23% MoNi and trehalose). Panel b illustrates the injection through a 21G needle and the lack of depot formation behavior of Formulation 3 (364 mg / mL BSA in sesame oil with trehalose but no copolymer). Panel c illustrates the injection through a 26G needle and the depot formation behavior of Formulation 7 (400 mg / mL BSA in triacetin with copolymer MoNi and trehalose). [Figure 4] Panels a-d illustrate angular frequency sweeps and flow sweeps of exemplary formulations. Panel a illustrates the angular frequency sweep of formulation 7 (400 mg / mL BSA in triacetin with MoNi). Panel b illustrates the flow sweep of formulation 7. Panel c illustrates the angular frequency sweep of formulation 6 (400 mg / mL BSA in triacetin without copolymer). Panel d illustrates the flow sweep of formulation 6. [Figure 5]Panels a-c illustrate the rheological and stability characterization of ball-milled BSA particles. Panel a illustrates the angular frequency sweep of i) 400 mg / mL BSA particles resuspended in triacetin and ii) 400 mg / mL BSA particles containing 5 wt% MoNi resuspended in triacetin. Panel a.iii) Comparative storage modulus of the suspensions at 10 rad / s. Panel b illustrates microscopy images of BSA with 5 wt% MoNi particles formed by i) 15 min of ball-milling or ii) spray-drying. The particles are resuspended in sesame oil for improved imaging. Panel c depicts SEC traces of a fresh BSA control, 15 min of ball-milled BSA without MoNi, and 15 min of ball-milled BSA with 5 wt% MoNi. PBS with sodium azide was used as the eluent. [Figure 6] Panels a-d illustrate angular frequency sweeps and flow sweeps of exemplary formulations formed by spray drying. Panel a illustrates the angular frequency sweep of formulation 11 (460 mg / mL BSA in triacetin with MoNi copolymer). Panel b illustrates the flow sweep of formulation 11. Panel c illustrates the angular frequency sweep of formulation 10 (460 mg / mL BSA in triacetin without copolymer). Panel d illustrates the flow sweep of formulation 10. Adding MoNi copolymer results in a 4000-fold decrease in formulation stiffness. [Figure 7]Panels a-d illustrate rheological and stability characterization of spray-dried particles. Panel a illustrates angular frequency sweeps of i) 460 mg / mL BSA particles resuspended in triacetin and ii) 460 mg / mL BSA particles containing 5 wt% MoNi resuspended in triacetin. Panel a.iii) Comparative storage modulus of the suspensions at 10 rad / s. Panel b depicts flow sweeps of 460 mg / mL BSA particles resuspended in triacetin and 460 mg / mL BSA particles containing 5 wt% MoNi resuspended in triacetin. Panel c shows SEM images (scale bar 5 μm) of particles and the resulting suspension in 460 mg / mL triacetin for particles formulated i) without and ii) with 5 wt% MoNi. Panel d depicts the SEC traces of fresh BSA control, spray-dried BSA without MoNi, and spray-dried BSA with 5 wt% MoNi. PBS with sodium azide is used as the eluent. [Figure 8] Panels a-b illustrate the effect of trehalose and copolymer MoNi on BSA stability after lyophilization (panel a) and milling (panel b). Panel a demonstrates that trehalose and MoNi content during lyophilization have little effect on the high molecular weight shoulder. Panel b further demonstrates that MoNi, regardless of trehalose content, reduces the size of the high molecular weight shoulder after 15 minutes of ball milling. [Figure 9]Panels a(i)–a(iii) and b show the results of evaluating exemplary particle compositions for protein stability using SEC. BSA protein compositions were prepared by spray drying or ball milling. SEC allows visualization of the BSA monomer and dimer peaks. Panel a(i): Full SEC traces of fresh, spray-dried, and ball-milled BSA. Panel a(ii): Dimer peaks of fresh, spray-dried, and ball-milled BSA. Panel a(iii): High molecular weight peaks of fresh, spray-dried particles (e.g., particles) with a 100:5 weight ratio of BSA, and ball-milled BSA. Panel b: BSA monomer fraction after spray drying and ball milling. These results suggest that spray drying is a milder process for particle formation than ball milling, but that MoNi is a useful protectant throughout the ball milling process. [Figure 10] Panels a-c and d(i)-d(ii) illustrate the evaluation of injection force and injection flow rate of exemplary compositions through different gauge needles. Panel a depicts the use of a force sensor connected to a syringe pump to quantify the injection force through a standard gauge needle using a 1 mL syringe. Panel b depicts injection force curves illustrating the injection force required to inject a suspension containing 460 mg / mL BSA and 23 mg / mL MoNi through a 27 G ½ inch needle at various flow rates. Panel c illustrates that the injection force is linear with the flow rate and varies with needle gauge. Panel d(i) shows injection force curves illustrating the injection force required to inject a suspension containing 460 mg / mL BSA and 23 mg / mL MoNi through a 27 G ½ inch needle and a 26 G ½ inch needle at 1 mL / min. Panel d(ii) illustrates that the injection force decreases with increasing needle gauge. Injection force comparisons with MoNi-free BSA formulations were not included because the MoNi-free formulations are not injectable through 26 or 27 G needles. [Figure 11]Panels a-c illustrate injection force studies of an exemplary composition through different gauge needles and after vertical storage. Panel a (i) further illustrates that injection force is linear with flow rate, and ii) varies with needle gauge. Panel b shows a picture of a syringe used for vertical storage of an exemplary composition for up to 120 hours. Panel c depicts the injection force of a 460 mg / mL BSA with 5% MoNi in triacetin composition on days 0 and 35 using a 26G ½ inch needle and a 1 mL / min flow rate. [Figure 12] Panels a-b show the results of a stress-aging test (30 minutes at 60°C) on an exemplary BSA formulation. Panel a depicts the SEC trace of the BSA formulation after stress aging. Panel b illustrates the corresponding BSA monomer fraction after stress aging conditions. BSA in a 20 mg / mL solution in PBS showed significant aggregation. BSA suspensions in triacetin showed minimal aggregation. A BSA suspension in triacetin containing 5 wt% MoNi in the suspension particles showed good stability with no high molecular weight peak beyond the dimer peak at 31 minutes. BSA in triacetin without MoNi in the particles shows a small high molecular weight peak at 16 minutes, but relatively good stability. [Figure 13] Panels a(i)-a(iii) show storage stability evaluations of exemplary compositions (460 mg / mL BSA formulations in triacetin with and without 5 wt. % MoNi in the particles of the suspension) stored for 48 hours at 4°C, 25°C, and 37°C. Panel a(i) depicts the complete SEC traces of the formulations after 48 hours. Panel a(ii) depicts the dimer peak for all formulations after 48 hours. Panel a(iii) depicts the high molecular weight peak for all formulations after 48 hours. Overall, all formulations showed good stability after 48 hours, with the majority of the light scattering signal coming from the monomer peak. Samples containing MoNi in the particles showed, on average, smaller dimer and high molecular weight peaks than samples without MoNi, suggesting improved storage stability. [Figure 14]Panels a(i)-a(iii) show storage stability evaluations of exemplary compositions (460 mg / mL BSA formulations in triacetin with and without 5 wt. % MoNi in the particles of the suspension) stored at 4°C, 25°C, and 37°C for 120 hours. Panel a(i) depicts the full SEC traces of the exemplary formulations after 120 hours. Panel a(ii) shows the dimer peak for all formulations after 120 hours. Panel a(iii) shows the high molecular weight peak for all formulations after 120 hours. BSA formulations containing MoNi in the particles stored in syringes for 120 hours did not exhibit particle settling. Overall, all formulations showed good stability after 120 hours, with the majority of the light scattering signal coming from the monomer peak. Samples containing MoNi in the particles showed, on average, a smaller high molecular weight peak than samples without MoNi, suggesting improved storage stability. [Figure 15] Shown from left to right are the results of a centrifuge study to evaluate suspension stability in the following vehicles: triacetin alone, triacetin with 20% benzyl benzoate, triacetin with 20% benzyl alcohol, and triacetin with 20% safflower oil. [Figure 16] Illustrates viscometer measurements of various liquid carriers. The graph illustrates how the viscosity of a higher viscosity non-solvent (propylene glycol (PG) or triacetin (T)) can be reduced by adding a lower viscosity additive such as benzyl alcohol (BA). [Figure 17]Panels a-d show that injection force measurements (1 mL / min through a 26 G ½-inch needle) show a significant decrease in injection force when DMAc is used in combination with triacetin as a nonsolvent. Panel a shows the plateau injection force of a triacetin suspension (100%) versus suspensions in various blends of triacetin:DMAc and triacetin:DMAc:BA. Panel b shows the injection force of a triacetin suspension versus DMAc content. Panel c shows the injection force over time of a triacetin suspension (100%) versus suspensions in various blends of triacetin:DMAc and triacetin:DMAc:BA. Panel d shows that the addition of polymeric MoNi affects the consistency of BSA powder dispersed in DMAc. The left image in panel d illustrates a formulation with 450 mg / mL BSA DMAc with MoNi. The right image in panel d illustrates a formulation with 450 mg / mL BSA in DMAc without the addition of MoNi. [Figure 18] Panels a-c show the results of in vivo administration of 4 mg of BSA to mice via either 200 injections of 20 mg / mL BSA in PBS (bolus injection) or 9 μL injections of 450 mg / mL BSA in triacetin (high-concentration paste / triacetin paste) (N≧3 per group). Panel a depicts the fluorescence signal after in vivo administration of 4 mg of BSA via either 200 injections of 20 mg / mL BSA in PBS (bolus injection) or 9 μL injections of 450 mg / mL BSA in triacetin (high-concentration paste / triacetin paste). Panel b shows the comparative administration volumes of the high-concentration paste and the 20 mg / mL bolus injection. Panel c shows the comparative half-lives of BSA subcutaneous absorption from the high-concentration paste and bolus administration. [Figure 19]Panels a-b show the results of in vivo administration of 9 μL of 450 mg / mL BSA in a 70:30 triacetin:DMAc suspension. Panel a is the fluorescence decay curve associated with in vivo administration of 4 mg of BSA via a 9 μL injection of 450 mg / mL BSA in 70:30 triacetin:DMAc. Panel b shows the half-life of subcutaneous absorption of paste in 100% triacetin, bolus injection (20 mg / mL BSA in PBS), and suspension in 70:30 triacetin:DMAc (N≧3 per group). [Figure 20] Panels a-b illustrate the products obtained after spray-drying formulations containing polysorbate 80 and Pluronic L-61. Panel a shows images of Falcon tubes containing comparative products recovered from spray-drying equal amounts of BSA with polysorbate 80 (left tube) and Pluronic L-61 (right tube). Panel b shows the spray-dryer glass components after spray-drying Pluronic L-61. As evident in the images, spray-drying Pluronic L-61 results in a sticky powder that adheres to the spray-dryer glass components, thus resulting in a low yield of spray-dried product (see the right tube in panel a, which shows almost no recovered product). [Figure 21] Panels a-c illustrate SEC traces of fresh BSA and BSA spray-dried with MoNi (BSA_MoNi), polysorbate 80 (BSA_Tw80), and without any additives (BSA_NoMoNi) (fresh BSA standard). Panel a shows that for all spray-dried samples, the monomer and dimer peaks appear similar. Panel b shows the difference between the spray-dried samples for the high molecular weight aggregate peak. Panel c shows that the area fraction of the high molecular weight peak is lowest for the fresh BSA standard, followed by BSA spray-dried in MoNi, BSA spray-dried with polysorbate 80, and finally, BSA spray-dried without any additives. [Figure 22]Panels a-c show SEC analysis of high molecular weight aggregates in aged protein pastes made with BSA, BSA with MoNi, and BSA with polysorbate 80. BSA aged at 20 mg / mL in PBS was used as a control. All samples were aged at 60°C for 60 minutes. Panel a shows the complete SEC trace, and panel b shows the difference between samples for the high molecular weight aggregate peak. Panel c shows the area percentage of the high molecular weight peak for each sample. [Figure 23] Panels a-c show injection force experiments (1 mL / min through a 26G 1 / 2 inch needle) using a 450 mg / mL BSA suspension in triacetin. BSA was spray dried with either polysorbate 80 (Tween 80) or MoNi in equimolar amounts (7.14 μmol per gram of BSA). Panel a shows the injection force (N) over time (seconds). Panel b shows the plateau injection force for formulations containing MoNi vs. polysorbate 80 (Tween 80). Panel c depicts SEM of the particle morphology of i) BSA-MoNi and ii) BSA-polysorbate 80 (Tween 80) particles (scale bar is 10 μm). [Figure 24] Panels a-b: Panel a depicts the comparative protein concentration predicted from density measurements and measured via nanodrop from known slurry volumes. Panel b illustrates the injection force as a function of concentration of BSA particles (with mole % matched MoNi or Tween 80) in 70 triacetin:30 DMAc. The injection force as a function of particle concentration is fitted to a particle jamming model. [Figure 25] Panels a-b show SEM images of BSA microparticle formulations. Panel a shows an SEM image of formulation 004A (smooth particles <5 μm). Panel b shows an SEM image of formulation 004B (smooth particles <5 μm). [Figure 26]Panels a-b show photographic images of BSA microparticle formulations suspended in triacetin and loaded into a 1 mL Schott syringe. Panel a shows a photographic image of a syringe loaded with formulation 004A (480 mg / mL) suspended in triacetin. Panel b shows a photographic image of a syringe loaded with formulation 004B (458 mg / mL) suspended in triacetin. [Figure 27] Panels a-b are graphs of the injection force (N) of BSA microparticle suspensions over 5 seconds at various flow rates. Panel a shows the injection force (N) over 5 seconds for BSA microparticle suspension 004A (480 mg / mL BSA) in triacetin injected at 2 mL / min, 4 mL / min, and 6 mL / min from a 1 mL Schott syringe (27 G thin-wall needle and V9519-coated plunger). Panel b shows the injection force (N) over 5 seconds for BSA microparticle suspension 004B (458 mg / mL BSA) in triacetin injected at 2 mL / min, 4 mL / min, and 6 mL / min from a 1 mL Schott syringe (27 G thin-wall needle and V9519-coated plunger). [Figure 28] 1 illustrates injection force measurements of 004A (480 mg / mL BSA) and 004B (458 mg / mL) microparticle suspensions in triacetin injected at different flow rates from a 1 mL Schott syringe (27G thin-walled needle and V9519 coated plunger). The graph shows that the injection force (N) of suspensions of 004A and 004B in triacetin is linear with flow rate. [Figure 29]Panels a-g illustrate the effective use of high-concentration suspension technology to deliver hIgG. Panel a depicts an SEM image of spray-dried IgG with MoNi particle morphology (scale bar is 10 μm). Panel b depicts SEC traces of a fresh hIgG control, spray-dried hIgG without MoNi, spray-dried hIgG with 5 wt% MoNi, and spray-dried hIgG with 25 wt% trehalose and 5 wt% MoNi. PBS with sodium azide is used as the eluent. Panel c shows a graph of the injection force as a function of concentration of hIgG microparticles with MoNi in triacetin. The injection force as a function of particle concentration is fitted to a particle jamming model. Panel d shows the injection force at 1 mL / min of a 350 mg / mL hIgG, 5 wt% MoNi suspension through a 26G ½-inch needle with DMAc, benzyl alcohol (BA), and benzyl benzoate (BB) non-solvent additives. Panel e(i) shows representative IVIS images of mice demonstrating subcutaneous absorption of fluorescently labeled hIgG administered via PBS bolus injection or high-concentration protein suspension. Panel e(ii) depicts a graph of the fluorescent signal within the subcutaneous space fitted to a monophasic exponential decay mode to identify the half-life of subcutaneous absorption. Panel f depicts a graph of the comparative half-life of subcutaneous absorption of hIgG administered via PBS bolus injection or UHC protein suspension formulated with triacetin. Panel g depicts a graph of the comparative volume of administration of bolus injection and high-concentration protein suspension. [Figure 30]Panels a-b. Panel a, i) shows the complete SEC traces of a fresh hIgG control, spray-dried hIgG with no additives, spray-dried hIgG with 1x MoNi (5 wt%), spray-dried hIgG with 2x MoNi (10 wt%), spray-dried hIgG with 25 wt% trehalose and 5 wt% MoNi, and spray-dried IgG with 30 wt% trehalose. Panel a, ii) shows the SEC trace of high molecular weight aggregates only. PBS with sodium azide was used as the eluent. Panel b shows a graph of the percent change in hIgG aggregates due to spray drying as a percent of total protein content. The baseline for determining percent change was determined by subtracting the starting percent aggregation of the fresh IgG control (3.9%) from all samples. [Figure 31] Panels a-b. Panel a depicts the injection force curve for a 1 mL / min injection of a 300 mg / mL IgG, 5 wt% MoNi suspension through a BD insulin syringe (28 G, 12.7 mm needle) with triacetin as the non-solvent, and panel b shows the corresponding plateau injection force graph. [Figure 32] Panels a-b show SEC traces of insulin before and after spray drying. Panel a shows the insulin SEC traces of spray-dried and pre-spray-dried insulin over a 22-minute period. Panel b is a zoomed-in portion of the SEC trace in panel a from 14 to 17 minutes. [Figure 33] Panels a-b show SEC traces of pramlintide before and after spray drying. (Panel a) shows the SEC traces of spray-dried and pre-spray-dried pramlintide over a 20-minute period. (Panel b) is a zoomed-in portion of the SEC trace in panel a from 14 to 17 minutes. [Figure 34] SEC traces of pramlintide ("pram"), insulin, and a control before and after spray drying are shown. [Figure 35]Panels a-b. SEM images of insulin and pramlintide spray-dried particles resuspended in triacetin (panel a), and trehalose and MoNi (95:5) particles (panel b). The particles in panel b contain neither insulin nor pramlintide, so proteins may affect particle morphology. [Figure 36] 1 shows insulin and pramlintide spray-dried particles resuspended in triacetin at a solids content of 350 mg / mL. The particle suspension flows like a liquid. [Figure 37] Panels a and b show the injection force of insulin pramlintide co-formulation in triacetin (350 mg / mL solids). Injection measurements were performed using a 1 mL syringe at 1 mL / min through a 26G ½ inch needle. [Figure 38] 1 is an image depicting insulin pramlintide co-formulation reconstituted in cell grade water. [Figure 39] Panels a-c show graphs of syringe force profiles for exemplary BSA formulations with various liquid carriers. Panel a depicts the syringe force profile for formulation 12 (benzyl benzoate as the liquid carrier). Panel b depicts the syringe force profile for formulation 13 (Miglyol 840 as the liquid carrier). Panel c depicts the syringe force profile for formulation 16 (ethyl oleate as the liquid carrier). [Figure 40] 1 is a graph showing the syringe force profile of exemplary BSA formulation 17 (triacetin:DMAc (75:25) as the liquid carrier). DETAILED DESCRIPTION OF THE INVENTION

[0009] 4. Detailed Description As summarized above, the present disclosure provides pharmaceutical compositions comprising particles suspended in a liquid carrier.The particles generally comprise a biopharmaceutical active agent that is administered to a subject who needs the administration of the biopharmaceutical active agent, and a polyacrylamide copolymer that provides benefits to the resulting composition, such as stability benefits, and / or provides formulation of the biopharmaceutical agent at a concentration suitable for administration via injection.Therefore, the pharmaceutical composition of the present disclosure can be referred to as an injectable pharmaceutical composition.

[0010] Reference will now be made in detail to particular embodiments of the disclosed subject matter. While the disclosed subject matter will be described in conjunction with the recited claims, it will be understood that the illustrated subject matter is not intended to limit the claims to the disclosed subject matter.

[0011] 4.1. Particles The pharmaceutical composition can include particles comprising a biopharmaceutical agent and a polyacrylamide-based copolymer.

[0012] The particles can be prepared from a precursor composition comprising a biopharmaceutical agent, a polyacrylamide-based copolymer, and one or more optional excipients (e.g., as described herein), and the composition can be subjected to various processes to prepare a particulate form of the precursor composition, including processes such as lyophilization, spray drying, freeze drying, spray freeze drying, milling, or combinations thereof. Once the particles are prepared, they are suspended in a liquid.

[0013] The particle size can depend on the particular composition and the preparation method selected. Characterization of the particle(s) size and / or dispersibility in the composition can be performed using methods such as optical microscope imaging.

[0014] The particles can include microparticles and / or nanoparticles.

[0015] In some embodiments, the particles have an average diameter of 100 microns or less, hi some embodiments, the particles have an average diameter of 95 microns or less, e.g., 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, or 5 microns or less.

[0016] In some embodiments, the particles have an average diameter of 0.01 to 100 microns, such as 0.01 to 100 microns, 0.01 to 50 microns, 0.01 to 20 microns, 0.01 to 10 microns, or 0.01 to 1 micron.

[0017] In some embodiments, the particles have an average diameter of 0.1 to 100 microns, such as 0.1 to 50 microns, or, for example, 0.1 to 20 microns, 0.1 to 10 microns, 10 to 20 microns, 20 to 30 microns, 30 to 40 microns, 40 to 50 microns, 50 to 60 microns, 60 to 70 microns, 70 to 80 microns, 80 to 90 microns, or 90 to 100 microns.

[0018] In some embodiments, the particles have an average diameter of 1 to 100 microns, e.g., 1 to 5 microns, 5 to 10 microns, 10 to 15 microns, 15 to 20 microns, 20 to 25 microns, 25 to 30 microns, 30 to 35 microns, 35 to 40 microns, 40 to 45 microns, 45 to 50 microns, 50 to 55 microns, 55 to 60 microns, 60 to 65 microns, 65 to 70 microns, 70 to 75 microns, 75 to 80 microns, 80 to 85 microns, 85 to 90 microns, 90 to 95 microns, and 95 to 100 microns.

[0019] In some embodiments, the particles have an average diameter of 1 to 50 microns, hi some embodiments, the particles have an average diameter of 10 to 20 microns, for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 microns.

[0020] The particles can be suspended in a liquid carrier (for example, as described herein). In some embodiments, the particle composition can be provided separately from the liquid carrier and formulated as a suspension in the liquid carrier before use. In some embodiments, a certain amount of the particle composition is provided in a suitable amount and concentration ready for use, already formulated as a suspension in the liquid carrier.

[0021] 4.1.1. Biopharmaceuticals The particles of the pharmaceutical composition also contain a biopharmaceutical agent. In some embodiments, the amount of biopharmaceutical agent present in the pharmaceutical composition is sufficient to provide a unit dose suitable for administration via injection. In some embodiments, the particles contain a single biopharmaceutical agent. In some embodiments, the particles contain a co-formulation of two or more biopharmaceutical agents.

[0022] In some embodiments, the particles are formulated in a weight percent of a volume of biopharmaceutical agent suitable for injection of a unit dose (e.g., SC or IM) into a patient in need thereof. In some embodiments, the biopharmaceutical agent has a high MW, e.g., a large biological agent. In some embodiments, a high MW biopharmaceutical agent is one having a MW of 20 kDa or greater, such as 30 kDa or greater, 40 kDa or greater, 50 kDa or greater, 60 kDa or greater, 70 kDa or greater, 80 kDa or greater, 90 kDa or greater, or 100 kDa or greater.

[0023] In some embodiments, the particles comprise about 20% or more by weight of the biopharmaceutical agent, e.g., about 25% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 65% or more, about 70% or more, about 75% or more, about 80% or more, about 85% or more, about 90% or more, or about 95% or more by weight of the biopharmaceutical agent. In some embodiments, the particles comprise about 90% to 95% by weight of the biopharmaceutical agent.

[0024] In some embodiments, the particles are formulated to contain a therapeutically effective amount of a biopharmaceutical agent, the biopharmaceutical agent having a MW of 10 kDa or less, such as 5 kDa or less. In some embodiments, the particles comprise 20 wt% or less of the biopharmaceutical agent, e.g., 15 wt% or less, 10 wt% or less, 9 wt% or less, 8 wt% or less, 7 wt% or less, 6 wt% or less, 5 wt% or less, 4 wt% or less, 3 wt% or less, 2 wt% or less, or 1 wt% or less of the biopharmaceutical agent. In some embodiments, the particles comprise about 1 wt% to 20 wt% of the biopharmaceutical agent, e.g., about 1 wt% to about 10 wt%. In some embodiments, the particles comprise 1 wt% to 20 wt% of the biopharmaceutical agent, e.g., 1 wt% to 10 wt%. In such cases, the remainder of the particle composition can be composed of one or more optional components, such as a polyacrylamide-based copolymer and / or a stabilizer (e.g., as described herein).

[0025] In some embodiments, the biopharmaceutical agent is a polypeptide. In some embodiments, the polypeptide is prone to aggregation in aqueous media. In some embodiments, the polypeptide is a protein. In some embodiments, the polypeptide is a peptide.

[0026] In some embodiments, the biopharmaceutical agent is selected from antibodies and fragments thereof, chimeric fusion proteins, cytokines, chemokines, hormones, vaccine antigens, cancer antigens, adjuvants, and combinations thereof. In some embodiments, the biopharmaceutical agent is a polypeptide.

[0027] In some embodiments, the biopharmaceutical agent is an antibody.

[0028] The term "antibody" is used herein in its broadest sense and includes a specific type of immunoglobulin molecule that contains one or more antigen-binding domains that specifically bind to an antigen or epitope. Antibodies specifically include, but are not limited to, full-length antibodies (e.g., intact immunoglobulins), antibody fragments, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), humanized fully human antibodies, chimeric antibodies, and single domain antibodies.

[0029] In some embodiments, the biopharmaceutical agent is a monoclonal antibody or a fragment thereof. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is monospecific, i.e., binds to a single antigen. In some embodiments, the monoclonal antibody is an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, an IgG4 antibody, an IgM antibody, an IgA antibody, or any hybrid thereof.

[0030] In some embodiments, the antibody is a chimeric antibody. The term "chimeric antibody" refers to an antibody that contains one or more regions from one antibody and one or more regions from one or more other antibodies.

[0031] In some embodiments, the antibody is multispecific, i.e., binds to multiple antigens, e.g., a bispecific antibody.

[0032] In some embodiments, the antibody is an antibody fragment. An "antibody fragment" includes a portion of an intact antibody, such as the antigen-binding or variable region of the intact antibody. Antibody fragments suitable for use in the present compositions include, for example, Fv fragments, Fab fragments, F(ab')2 fragments, Fab' fragments, scFv (sFv) fragments, and scFv-Fc fragments.

[0033] In some embodiments, the biopharmaceutical agent is a chimeric protein. In some embodiments, the chimeric protein is a recombinant fusion protein. A chimeric protein can include two or more domains connected via an optional linker or spacer. In some embodiments, the chimeric protein comprises an antibody fragment, such as Fc or a fragment or variant thereof. In some embodiments, the chimeric protein comprises an antibody fragment fused to a protein domain, e.g., a protein domain that specifically binds to a therapeutic target of the biopharmaceutical agent. In some embodiments, the chimeric protein is an Fc fusion protein.

[0034] In some embodiments, the antibody is a single domain antibody, e.g., a camelid antibody, or a nanobody.

[0035] In some embodiments, the antibody is an antibody-drug conjugate, e.g., an antibody conjugated to one or more heterologous molecules. The heterologous molecule can be a small molecule (e.g., an organic compound having a molecular weight of less than 1000, 900, 800, 700, 600, or 500 daltons). In some embodiments, the heterologous molecule is a cytotoxic agent, a chemotherapeutic agent, or a cytostatic agent.

[0036] In some embodiments, the antibody is a bispecific antibody immunoconjugate.

[0037] In some embodiments, the antibody is an antibody fragment. In some embodiments, the antibody comprises one or more single-chain variable fragments (scFv) of a monoclonal antibody. In some embodiments, the monoclonal antibody is a humanized antibody, a human antibody, a murine antibody, or a chimeric (murine / human) antibody.

[0038] The antibodies can be targeted to a variety of target proteins, for example, therapeutic target proteins.

[0039] In some embodiments, the antibody has a molecular weight of about 100 kDa to about 200 kDa, e.g., about 120 kDa to about 180 kDa. In some embodiments, the antibody has a molecular weight of about 150 kDa. In some embodiments, the antibody has a molecular weight of 100 kDa to 200 kDa, e.g., 120 kDa to 180 kDa. In some embodiments, the antibody has a molecular weight of about 150 kDa. In some embodiments, the antibody has a molecular weight of 100 kDa or less, such as 40 kDa to 80 kDa, e.g., about 50 kDa.

[0040] In some embodiments, the particles comprise 5% or more antibody by weight, e.g., 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 95% or more antibody by weight. In some embodiments, the composition comprises 95% or less, 90% or less, or 80% or less antibody by weight. In some embodiments, the antibody is a monoclonal antibody or a fragment thereof.

[0041] In some embodiments, the particles comprise about 60% to about 99% by weight of antibody, e.g., about 65% to about 95% by weight, about 70% to about 95% by weight, about 75% to about 95% by weight, about 80% to about 95% by weight, about 85% to about 95% by weight, or about 90% to about 95% by weight of antibody. In some embodiments, the particles comprise about 90% or about 95% by weight of antibody. In some embodiments, the particles comprise 60% to 99% by weight of antibody, e.g., 65% to 95% by weight, 70% to 95% by weight, 75% to 95% by weight, 80% to 95% by weight, 85% to 95% by weight, or 90% to 95% by weight of antibody. In some embodiments, the antibody is a monoclonal antibody or a fragment thereof.

[0042] In some embodiments, the biopharmaceutical agent is insulin.

[0043] The term "insulin" refers to a hormone produced by beta cells in pancreatic islets that regulates the amount of glucose in the blood. Many eukaryotic organisms, including humans, primates, pigs, cattle, cats, dogs, and rodents, produce insulin. Therefore, as used herein, "insulin" includes insulin and analogs thereof produced by humans, as well as insulin and analogs thereof produced by other eukaryotic organisms, including, but not limited to, primates, pigs, cattle, cats, dogs, and rodents, and also includes recombinant, purified, or synthetic insulin or insulin analogs with similar function and structure, unless otherwise specified. Human insulin protein consists of 51 amino acids and has a molecular weight of approximately 5.8 kilodaltons (kDa). Human insulin is a heterodimer of an A chain and a B chain connected by a disulfide bond.

[0044] Insulin can be isolated from pancreatic islet extracts of insulin-producing animals or can be recombinantly expressed in suitable expression systems such as E. coli, yeast, insect cells, and mammalian cells (e.g., Chinese hamster ovary (CHO) cells). Their specific pharmacokinetic and pharmacodynamic (PK / PD) properties (e.g., duration of action, maximum observed concentration (C max Depending on the dosage, time to onset, and area under the curve (AUC), insulin can be further characterized as rapid-acting insulin, short-acting insulin, intermediate-acting insulin, long-acting insulin, and premixed insulin.

[0045] Insulin also includes monomeric and oligomeric forms, such as dimeric and hexameric forms. Insulin can exist as a monomer as it circulates in the plasma and binds to its receptor while in monomeric form. Insulin preparations (or insulin analog preparations) containing a preponderance of monomeric and dimeric protein molecules typically have a strong tendency to aggregate and form inactive fibrils. Because insulin hexamers are too large to be absorbed, hexameric insulin preparations must be broken down into dimers or monomers before the insulin can be absorbed and function in the body. The active form of insulin in the bloodstream is the monomeric form.

[0046] In some embodiments, the particles comprise about 0.5% to about 20%, about 1% to about 20%, about 1% to about 15%, about 1% to about 10%, about 1% to about 9%, or about 1% to about 8% insulin or analogue thereof by weight. In some embodiments, the particles comprise about 7%, about 8%, about 9%, or about 10% insulin or analogue thereof by weight. In some embodiments, the particles comprise 1% to 20%, 1% to 15%, 1% to 10%, 1% to 9%, or 1% to 8% insulin or analogue thereof by weight. In some embodiments, the particles comprise 7%, 8%, 9%, or 10% insulin or analogue thereof by weight.

[0047] In some embodiments, the insulin or analog thereof is selected from the group consisting of insulin lispro, HUMALOG® (rapid-acting insulin lispro), insulin glargine, LANTUS® (insulin glargine), insulin detemir, LEVEMIR® (insulin detemir), ACTRAPID® (rapid-acting human insulin), modern insulin, NOVORAPID® (insulin aspart), VELOSULIN® (human insulin), HUMULIN® M3 (a mixture of soluble insulin and isophane insulin called biphasic isophane insulin), HYPURIN® (neutral bovine insulin), INSUMAN® (recombinant human insulin), INSULATARD® (long-acting isophane human insulin), MIXTARD® 30 (a mixture of 30% soluble insulin and 70% isophane insulin), MIXTARD® 40 (a mixture of 40% soluble insulin and 60% isophane insulin), MIXTARD (R) 50 (a mixture of 50% soluble insulin and 50% isophane insulin), insulin aspart, insulin glulisine, insulin isophane, insulin degludec, insulin icodec, long-acting zinc insulin, NOVOLIN® R (human insulin), HUMULIN® R (human insulin), HUMULIN® Regular U-500 (concentrated regular insulin), NOVOLIN® N (intermediate-acting human insulin), HUMULIN® N ( intermediate-acting human insulin), RELION® (commercial brands of NOVOLIN® R, NOVOLIN® N, and NOVOLIN® 70 / 30), AFREZZA® (rapid-acting inhaled insulin), HUMULIN® 70 / 30 (a mixture of 70% human insulin isophane suspension and 30% human insulin injection), NOVOLIN® 70 / 30 (a mixture of 70% NPH, human insulin isophane suspension, and 30% regular human insulin injection),The insulin or analog thereof is selected from NOVOLOG® 70 / 30 (a mixture of 70% insulin aspart protamine suspension and 30% insulin aspart injection), HUMULIN® 50 / 50 (a mixture of 50% human insulin isophane suspension and 50% human insulin injection), HUMALOG® Mix 75 / 25 (a mixture of 75% insulin lispro protamine suspension and 25% insulin lispro injection), insulin aspart protamine-insulin aspart, insulin lispro protamine-insulin lispro, human insulin NPH-human insulin regular, insulin degludec-insulin aspart, and combinations thereof. In some embodiments, the insulin or analog thereof is human insulin or recombinant human insulin. In some embodiments, the insulin or analog thereof is non-human (e.g., primate, porcine, bovine, feline, canine, or rodent) insulin or recombinant non-human insulin. In some embodiments, the insulin or analog thereof is purified or synthetic insulin. In some embodiments, the insulin or analog thereof is selected from rapid-acting insulin, short-acting insulin, intermediate-acting insulin, long-acting insulin, and premixed insulin. In some embodiments, the insulin or analog thereof is insulin lispro. In some embodiments, the insulin or analog thereof is insulin aspart. In some embodiments, the insulin or analog thereof is recombinant human insulin.

[0048] In some embodiments, the biopharmaceutical agent is a peptide or peptide analog. The formulation techniques described herein should be particularly useful for preparing shelf-stable, injectable pharmaceutical compositions of various therapeutic peptides, including, but not limited to, glucagon, glucagon-like peptide-1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP), GLP-1 receptor agonist, GLP-2, adrenocorticotropic hormone (ACTH), leuprolide, hirudin, insulin, pramlintide, exendin, exenatide, gastric inhibitory peptide, calcitonin, calcitonin gene-related peptide, amylin, adrenomedrin, angiotensin, immunogenic peptides (e.g., peptides or peptide complexes derived from viruses, bacteria, or any eukaryotic organism or cell thereof), and the like, and analogs thereof.

[0049] In some embodiments, the biopharmaceutical agent is a glucagon peptide, glucagon analog, glucagon mimetic, or a salt thereof.

[0050] 4.1.1.1 Additional activators The particles of the present disclosure can include a combination of a biopharmaceutical agent and one or more additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents are useful for treating a disease or condition targeted by the biopharmaceutical agent. In some embodiments, administration of the agents from a pharmaceutical composition can provide delivery to cells in a subject's body so that they simultaneously exert their biological or therapeutic effects. In some embodiments, the agents within the particles are co-formulated to provide substantially similar pharmacokinetic profiles.

[0051] In some embodiments, the one or more additional therapeutic agents are active agents that provide a synergistic effect with the biopharmaceutical agent, hi some embodiments, the additional therapeutic agent is a small molecule drug.

[0052] In some embodiments, the stable formulations and particles used in accordance with the present disclosure comprise co-formulations or mixtures of biopharmaceutical agents of the type described herein, such as at least one peptide, at least one small molecule, and combinations thereof.

[0053] In some embodiments, the particles are co-formulated to contain a first biopharmaceutical agent that is a protein and a second biopharmaceutical agent that is a peptide. In some embodiments, the particles are co-formulated to contain a first and a second biopharmaceutical agent that are peptides. In some embodiments, the particles are co-formulated to contain a first biopharmaceutical agent that is a peptide hormone or its analog, and a second therapeutic agent that is a small molecule. In some embodiments, the second therapeutic agent is a steroid. In some embodiments, the co-formulation is a mixture of a first batch of particles containing a biopharmaceutical that is a protein and a second batch of particles containing a biopharmaceutical that is a peptide.

[0054] In some embodiments, the particles are co-formulated to contain a first biopharmaceutical agent that is insulin and a second biopharmaceutical agent that is a peptide. In some embodiments, the co-formulation is a mixture of a first batch of particles containing insulin and a second batch of particles containing a peptide. In some embodiments, the peptide is pramlintide.

[0055] The amounts of active agent(s) that can be combined with each other and with the particulate carrier material to produce a dosage form will vary depending on the subject and the particular mode of administration required.

[0056] 4.1.2. Polyacrylamide copolymers The particles provided in this disclosure can include a polyacrylamide-based copolymer formulated with a biopharmaceutical agent of interest.

[0057] Polyacrylamide-based copolymers of interest and that can be used in the particles and particle suspensions described herein include those described in WO 2021 / 211976 and WO 2023 / 230046, the disclosures of which are incorporated herein by reference.

[0058] In some embodiments, the polyacrylamide-based copolymer contains a water-soluble carrier monomer and a functional dopant monomer. In some embodiments, the polyacrylamide-based copolymer is amphiphilic.

[0059] In some embodiments, the polyacrylamide-based copolymer comprises a non-ionic water-soluble acrylamide monomer and a functional dopant monomer, hi some embodiments, the polyacrylamide-based copolymer further comprises a functional acrylamide dopant monomer selected from a hydrophobic functional acrylamide dopant monomer, an aromatic functional acrylamide dopant monomer, a hydrogen-bonding functional acrylamide dopant monomer, and an ionic functional acrylamide dopant monomer.

[0060] The polyacrylamide-based copolymers of the present disclosure contain a water-soluble carrier monomer. In some embodiments, the water-soluble carrier monomer is non-ionic. In some embodiments, the water-soluble carrier monomer is selected from N-(3-methoxypropoyl)acrylamide (MPAM), 4-acryloylmorpholine (MORPH), N,N-dimethylacrylamide (DMA), N-hydroxyethylacrylamide (HEAM), and acrylamide (AM), or a combination thereof. In some embodiments, the water-soluble carrier monomer is selected from MPAM and MORPH. In some embodiments, the water-soluble carrier monomer is N-(3-methoxypropoyl)acrylamide (MPAM). In some embodiments, the water-soluble carrier monomer is 4-acryloylmorpholine (MORPH). In some embodiments, the water-soluble carrier monomer is N,N-dimethylacrylamide (DMA). In some embodiments, the water-soluble carrier monomer is N-hydroxyethylacrylamide (HEAM). In some embodiments, the water-soluble carrier monomer is acrylamide (AM). In some embodiments, the copolymer comprises a water-soluble carrier monomer selected from N-(3-methoxypropoyl)acrylamide (MPAM) and 4-acryloylmorpholine (MORPH).

[0061] The polyacrylamide-based copolymers of the present disclosure also include 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-N-diethylacrylamide (DEA), N-tert-butylacrylamide (TBA), N-phenylacrylamide (PHE), and combinations thereof. In some embodiments, the polyacrylamide-based copolymer includes a 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 comprises 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 comprises a hydrogen-bonding functional acrylamide dopant monomer. In some embodiments, the hydrogen-bonding functional acrylamide dopant monomer is N-[tris(hydroxymethyl)-methyl]acrylamide (TRI). In some embodiments, the polyacrylamide-based copolymer 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 ionically 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). In some embodiments, the functional dopant monomer is N,N-diethylacrylamide (DEA).

[0062] In some embodiments, the polyacrylamide-based copolymer comprises a water-soluble carrier monomer selected from N-(3-methoxypropoyl)acrylamide (MPAM), 4-acryloylmorpholine (MORPH), N,N-dimethylacrylamide (DMA), N-hydroxyethylacrylamide (HEAM), and acrylamide (AM), and 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-N-diethylacrylamide (DEA), N-tert-butylacrylamide (TBA), and N-phenylacrylamide (PHE).

[0063] In some embodiments, the water soluble carrier monomer is N-(3-methoxypropoyl)acrylamide (MPAM). In some embodiments, the water soluble carrier monomer is N-(3-methoxypropoyl)acrylamide (MPAM) and the functional dopant monomer is N-phenylacrylamide (PHE).

[0064] 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-isopropylacrylamide (NIP) or N-phenylacrylamide (PHE).

[0065] In some embodiments, the water-soluble carrier monomer is N,N-dimethylacrylamide (DMA). In some embodiments, the water-soluble carrier monomer comprises N,N-dimethylacrylamide (DMA) and 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-N-diethylacrylamide (DEA), N-tert-butylacrylamide (TBA), N-phenylacrylamide (PHE), and combinations thereof.

[0066] In some embodiments, the water-soluble carrier monomer is N-hydroxyethylacrylamide (HEAM). In some embodiments, the water-soluble carrier monomer is N-hydroxyethylacrylamide (HEAM) and the functional dopant monomer is selected from 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), N-phenylacrylamide (PHE), and combinations thereof.

[0067] 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 selected from 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), N-phenylacrylamide (PHE), and combinations thereof.

[0068] In some embodiments, the polyacrylamide-based copolymer comprises N-(3-methoxypropoyl)acrylamide (MPAM) or 4-acryloylmorpholine (MORPH) as the water-soluble carrier monomer, and the functional dopant monomer comprises one or more of 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).

[0069] In some embodiments, the polyacrylamide-based copolymer comprises N,N-dimethylacrylamide (DMA), N-hydroxyethylacrylamide (HEAM), or acrylamide (AM) as the water-soluble carrier monomer, and the functional dopant monomer is selected from 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), N-phenylacrylamide (PHE), and combinations thereof.

[0070] In some embodiments, the amount of functional dopant monomer used in the copolymerization reaction is designed to maximize dopant loading while obtaining a functional copolymer with 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 contains about 2% to about 30% by weight of the functional dopant monomer, e.g., about 5% to about 30% by weight, about 10% to about 30% by weight, about 15% to about 30% by weight, about 20% to about 30% by weight, about 25% to about 30% by weight, about 2% to about 25% by weight, about 5% to about 25% by weight, about 10% to about 25% by weight, about 15% to about 25% by weight, about 15% to about 25% by weight, about 20% to about 30% by weight, about 25% to about 30% by weight, about 2% to about 25% by weight, about 5% to about 25% by weight, about 10% to about 25% by weight, about 15% to about 25% by weight, about 20% to about 30% by weight, about 25% to about 30% by weight, about 25% to about 25 ... In some embodiments, the polyacrylamide-based copolymer comprises about 2 wt%, about 5 wt%, about 8 wt%, about 10 wt%, about 12 wt%, about 15 wt%, about 18 wt%, about 20 wt%, about 22 wt%, about 25 wt%, about 20 wt%, about 25 wt%, about 2 wt% to about 25 wt%, about 2 wt% to about 20 wt%, about 5 wt% to about 20 wt%, about 10 wt% to about 20 wt%, about 15 wt%, about 2 wt% to about 15 wt%, about 5 wt% to about 15 wt%, about 10 wt%, about 2 wt% to about 10 wt%, or about 2 wt% to about 5 wt% of the functional dopant monomer. In some embodiments, the polyacrylamide-based copolymer comprises about 2 wt%, about 5 wt%, about 8 wt%, about 10 wt%, about 12 wt%, about 15 wt%, about 18 wt%, about 20 wt%, about 22 wt%, about 25 wt%, about 28 wt%, or about 30 wt% of the functional dopant monomer. In some embodiments, the polyacrylamide-based copolymer comprises 2% to 30% by weight of the functional dopant monomer, e.g., 5% to 30% by weight, 10% to 30% by weight, 15% to 30% by weight, 20% to 30% by weight, 25% to 30% by weight, 2% to 25% by weight, 5% to 25% by weight, 10% to 25% by weight, 15% to 25% by weight, 20% to 25% by weight, 2% to 20% by weight, 5% to 20% by weight, 10% to 20% by weight, 15% to 20% by weight, 2% to 15% by weight, 5% to 15% by weight, 10% to 15% by weight, 2% to 10% by weight, 5% to 10% by weight, or 2% to 5% 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 a functional dopant monomer.

[0071] In some embodiments, the polyacrylamide copolymer comprises about 70% to about 98% by weight of a water-soluble carrier monomer, e.g., about 75% to about 98% by weight, about 80% to about 98% by weight, about 85% to about 98% by weight, about 90% to about 98% by weight, about 95% to about 98% by weight, about 70% to about 95% by weight, about 75% to about 95% by weight, about 80% to about 95% by weight, about 85% to about 98% by weight The polyacrylamide copolymer comprises about 95%, about 90% to about 95%, about 70% to about 90%, about 75% to about 90%, about 80% to about 90%, about 85% to about 90%, about 70% to about 85%, about 75% to about 85%, about 80% to about 85%, about 70% to about 80%, about 75% to about 80%, or about 70% to about 75% by weight of the water-soluble carrier monomer. In some embodiments, the polyacrylamide 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 the water-soluble carrier monomer. In some embodiments, the polyacrylamide-based copolymer comprises 70% to 98% by weight of the water-soluble carrier monomer, e.g., 75% to 98% by weight, 80% to 98% by weight, 85% to 98% by weight, 90 to 98% by weight, 95 to 98% by weight, 70% to 95% by weight, 75% to 95% by weight, 80% to 95% by weight, 85% to 95% by weight, 90% to 95% by weight, 70% to 90% by weight, 75% to 90% by weight, 80% to 90% by weight, 85% to 90% by weight, 70% to 85% by weight, 75% to 85% by weight, 80% to 85% by weight, 70% to 80% by weight, 75% to 80% by weight, or 70% to 75% by weight of the 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.

[0072] In some embodiments, the polyacrylamide-based copolymer comprises about 70% to about 98% by weight of the water-soluble carrier monomer and about 2% to about 30% by weight of the functional dopant monomer. For example, the polyacrylamide-based copolymer may comprise about 70% to about 98%, about 70% to about 95%, about 70% to about 80%, about 80% to about 95%, about 90% to about 98%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 98% by weight of the water-soluble carrier monomer and about 2% to about 30%, about 5% to about 25%, about 5% to about 20%, about 2% The functional dopant monomer may be contained in an amount of about 2% by weight, about 5% by weight to about 17% by weight, about 5% by weight to about 10% by weight, about 10% by weight to about 15% by weight, about 15% by weight to about 20% by weight, about 20% by weight to about 25% by weight, or about 25% by weight to about 30% by weight, about 2% by weight, about 5% by weight, about 8% by weight, about 10% by weight, about 12% by weight, about 15% by weight, about 18% by weight, about 20% by weight, about 22% by weight, about 25% by weight, about 28% by weight, or about 30% by weight.

[0073] 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 comprise 70% to 98%, 70% to 95%, 70% to 80%, 80% to 95%, 90% to 98%, 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 20%, ... % to 5 wt%, 2 wt% to 17 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 a functional dopant monomer.

[0074] In some embodiments, the polyacrylamide-based copolymer comprises about 2% to about 30% by weight of the functional dopant monomer NIP. In some embodiments, the polyacrylamide-based copolymer comprises about 5% to about 30% by weight of the functional dopant monomer NIP. In some embodiments, the polyacrylamide-based copolymer comprises about 10% to about 28% by weight of the functional dopant monomer NIP. In some embodiments, the polyacrylamide-based copolymer comprises about 5% to about 26% by weight of the functional dopant monomer NIP. In some embodiments, the polyacrylamide-based copolymer comprises about 5% to about 10% by weight of the functional dopant monomer NIP. In some embodiments, the polyacrylamide-based copolymer comprises about 10% to about 15% by weight of the functional dopant monomer NIP. In some embodiments, the polyacrylamide-based copolymer comprises about 15% to about 20% by weight of the functional dopant monomer NIP. In some embodiments, the polyacrylamide-based copolymer comprises about 20% to about 26% by weight of the functional dopant monomer NIP.

[0075] In some embodiments, the polyacrylamide-based copolymer comprises 2% to 30% by weight of a functional dopant monomer NIP, such as 5% to 30% by weight, 10% to 28% by weight, 5% to 26% by weight, 5% to 10% by weight, 10% to 15% by weight, 15% to 20% by weight, or 20% to 26% by weight of a functional dopant monomer NIP.

[0076] In some embodiments, the polyacrylamide-based copolymer comprises from about 2% to about 30% by weight of a functional dopant monomer PHE. In some embodiments, the polyacrylamide-based copolymer comprises from about 5% to about 30% by weight of a functional dopant monomer PHE. In some embodiments, the polyacrylamide-based copolymer comprises from about 10% to about 28% by weight of a functional dopant monomer PHE. In some embodiments, the polyacrylamide-based copolymer comprises from about 5% to about 26% by weight of a functional dopant monomer PHE. In some embodiments, the polyacrylamide-based copolymer comprises from about 5% to about 10% by weight of a functional dopant monomer PHE. In some embodiments, the polyacrylamide-based copolymer comprises from about 10% to about 15% by weight of a functional dopant monomer PHE. In some embodiments, the polyacrylamide-based copolymer comprises from about 15% to about 20% by weight of a functional dopant monomer PHE. In some embodiments, the polyacrylamide-based copolymer comprises about 20% to about 26% by weight of the functional dopant monomer PHE.

[0077] In some embodiments, the polyacrylamide-based copolymer comprises 2% to 30% by weight of a functional dopant monomer PHE, such as 5% to 30% by weight, 10% to 28% by weight, 5% to 26% by weight, 5% to 10% by weight, 10% to 15% by weight, 15% to 20% by weight, or 20% to 26% by weight of a functional dopant monomer PHE.

[0078] In some embodiments, the polyacrylamide-based copolymer comprises about 2% to about 30% by weight of the functional dopant monomer DEA. In some embodiments, the polyacrylamide-based copolymer comprises about 5% to about 30% by weight of the functional dopant monomer DEA. In some embodiments, the polyacrylamide-based copolymer comprises about 10% to about 28% by weight of the functional dopant monomer DEA. In some embodiments, the polyacrylamide-based copolymer comprises about 5% to about 26% by weight of the functional dopant monomer DEA. In some embodiments, the polyacrylamide-based copolymer comprises about 5% to about 10% by weight of the functional dopant monomer DEA. In some embodiments, the polyacrylamide-based copolymer comprises about 10% to about 15% by weight of the functional dopant monomer DEA. In some embodiments, the polyacrylamide-based copolymer comprises about 15% to about 20% by weight of the functional dopant monomer DEA. In some embodiments, the polyacrylamide-based copolymer comprises about 20% to about 26% by weight of the functional dopant monomer DEA.

[0079] In some embodiments, the polyacrylamide-based copolymer comprises 2% to 30% by weight of the functional dopant monomer DEA, such as 5% to 30% by weight, 10% to 28% by weight, 5% to 26% by weight, 5% to 10% by weight, 10% to 15% by weight, 15% to 20% by weight, or 20% to 26% by weight of the functional dopant monomer DEA.

[0080] In some embodiments, the polyacrylamide-based copolymer comprises MORPH as a water-soluble carrier monomer and about 2% to about 30% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the copolymer comprises MORPH and about 5% to about 30% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the copolymer comprises MORPH and about 10% to about 28% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the copolymer comprises MORPH and about 5% to about 26% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the copolymer comprises MORPH and about 5% to about 10% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the copolymer comprises MORPH and about 10% to about 15% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the copolymer comprises MORPH and about 15% to about 20% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the copolymer comprises MORPH and about 20% to about 25% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the copolymer comprises MORPH and about 25% to about 30% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the copolymer comprises MORPH and about 20% to about 28% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the copolymer comprises MORPH and about 21 wt% of a functional dopant monomer selected from NIP, PHE, and DEA, hi some embodiments, the copolymer comprises MORPH and about 22 wt% of a functional dopant monomer selected from NIP, PHE, and DEA.In some embodiments, the copolymer comprises MORPH and about 23 wt% of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the copolymer comprises MORPH and about 24 wt% of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the copolymer comprises MORPH and about 25 wt% of a functional dopant monomer selected from NIP, PHE, and DEA.

[0081] In some embodiments, the polyacrylamide copolymer comprises MORPH as a water-soluble carrier monomer and 2 wt% to 30 wt% of a functional dopant monomer selected from NIP, PHE, and DEA, such as 5 wt% to 30 wt%, 10 wt% to 28 wt%, 5 wt% to 26 wt%, 5 wt% to 10 wt%, 10 wt% to 15 wt%, 15 wt% to 20 wt%, 20 wt% to 25 wt%, 25 wt% to 30 wt%, or 20 wt% to 28 wt% of the functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the copolymer comprises 21 wt% of a functional dopant monomer selected from NIP, PHE, and DEA, such as 22, 23, 24, or 25 wt% of the functional dopant monomer selected from NIP, PHE, and DEA.

[0082] In some embodiments, the polyacrylamide-based copolymer comprises MPAM as a water-soluble carrier monomer and about 2% to about 16% by weight of a functional dopant monomer selected from NIP, PHE, or DEA. In some embodiments, the polyacrylamide-based copolymer comprises MPAM as a water-soluble carrier monomer and about 5% to about 15% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the polyacrylamide-based copolymer comprises MPAM as a water-soluble carrier monomer and about 6% to about 10% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the polyacrylamide-based copolymer comprises MPAM as a water-soluble carrier monomer and about 7% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the polyacrylamide-based copolymer comprises MPAM as a water-soluble carrier monomer and about 8 wt.% of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the polyacrylamide-based copolymer comprises MPAM as a water-soluble carrier monomer and about 9 wt.% of a functional dopant monomer selected from NIP, PHE, and DEA.

[0083] In some embodiments, the polyacrylamide-based copolymer comprises MPAM as a water-soluble carrier monomer and 2% to 16% by weight of a functional dopant monomer selected from NIP, PHE, and DEA, such as 5% to 15% by weight, or 6% to 10% by weight of a functional dopant monomer selected from NIP, PHE, and DEA. In some embodiments, the polyacrylamide-based copolymer comprises MPAM as a water-soluble carrier monomer and 7% by weight of a functional dopant monomer selected from NIP, PHE, and DEA, such as 8% or 9% by weight of a functional dopant monomer selected from NIP, PHE, and DEA.

[0084] In some embodiments, the polyacrylamide-based copolymer further comprises TRI, AMP, TMA, or TBA as a functional dopant monomer. In some embodiments, the AMP, TMA, or TBA functional dopant monomer is present at about 2% to about 16% by weight of the copolymer. In some embodiments, the TRI, AMP, TMA, or TBA functional dopant monomer is present at about 5% to about 15% by weight of the copolymer. In some embodiments, the TRI, AMP, TMA, or TBA functional dopant monomer is present at about 6% to about 14% by weight of the copolymer. In some embodiments, the TRI, AMP, TMA, or TBA functional dopant monomer is present at about 12% to about 15% by weight of the copolymer. In some embodiments, the TRI, AMP, TMA, or TBA functional dopant monomer is present at about 2% to about 5% by weight of the copolymer. In some embodiments, the TRI, AMP, TMA, or TBA functional dopant monomer is present at about 5% to about 10% by weight of the copolymer.

[0085] In some embodiments, the polyacrylamide-based copolymer further comprises AMP, TMA, or TBA as a functional dopant monomer in an amount of 2% to 16%, such as 5% to 15%, 6% to 14%, or 12% to 15% by weight of the copolymer. In some embodiments, the TRI, AMP, TMA, or TBA functional dopant monomer is present at about 2% to about 5% by weight of the copolymer. In some embodiments, the TRI, AMP, TMA, or TBA functional dopant monomer is present at about 5% to about 10% by weight of the copolymer.

[0086] In some embodiments, the polyacrylamide-based copolymer comprises 70% to 85% by weight of MORPH as the water-soluble carrier monomer and 15% to 30% by weight of NIP as the functional dopant monomer. In some embodiments, the polyacrylamide-based copolymer comprises 74% to 80% by weight of MORPH as the water-soluble carrier monomer and 20% to 26% by weight of NIP as the functional dopant monomer. In some embodiments, the polyacrylamide-based copolymer comprises 70% by weight of MORPH and 30% by weight of NIP. In some embodiments, the polyacrylamide-based copolymer comprises 71% by weight of MORPH and 29% by weight of NIP. In some embodiments, the polyacrylamide-based copolymer comprises 72% by weight of MORPH and 28% by weight of NIP. In some embodiments, the polyacrylamide-based copolymer comprises 73% by weight of MORPH and 27% by weight of NIP. In some embodiments, the polyacrylamide-based copolymer comprises 74 wt% MORPH and 26 wt% NIP. In some embodiments, the polyacrylamide-based copolymer comprises 75 wt% MORPH and 25 wt% NIP. In some embodiments, the polyacrylamide-based copolymer comprises 76 wt% MORPH and 24 wt% NIP. In some embodiments, the polyacrylamide-based copolymer comprises 77 wt% MORPH and 23 wt% NIP. In some embodiments, the polyacrylamide-based copolymer comprises 78 wt% MORPH and 22 wt% NIP. In some embodiments, the polyacrylamide-based copolymer comprises 79 wt% MORPH and 21 wt% NIP. In some embodiments, the polyacrylamide-based copolymer comprises 80 wt% MORPH and 20 wt% NIP. In some embodiments, the polyacrylamide-based copolymer comprises 81 wt% MORPH and 19 wt% NIP. In some embodiments, the polyacrylamide-based copolymer comprises 82% by weight MORPH and 18% by weight NIP, hi some embodiments, the polyacrylamide-based copolymer comprises 83% by weight MORPH and 17% by weight NIP.In some embodiments, the polyacrylamide-based copolymer comprises 84% ​​by weight MORPH and 16% by weight NIP, hi some embodiments, the polyacrylamide-based copolymer comprises 85% by weight MORPH and 15% by weight NIP.

[0087] In some embodiments, the degree of polymerization (DP) of the polyacrylamide 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 DP of the copolymer is about 100.

[0088] 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, 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.

[0089] In some embodiments, the molecular weight of the polyacrylamide copolymer is from about 1,000 g / mol to about 40,000 g / mol, e.g., 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, or from about 1,000 g / mol to about 1,000 g / mol. 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 2,000 g / mol to about 10,000 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, about 3,0 00 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 4,000 g / mol to about 10,000 g / mol, about 4, 5,000 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 5,000 g / mol to about 10,000 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 40,0 00 g / mol, about 7,000 g / mol to about 35,000 g / mol, about 7,000 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 10,000 g / mol to about 30,000 g / mol, about 1 0,000 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 to about 40,000 g / mol, about 20, In some embodiments, the molecular weight of the copolymer is about 1,000 to about 30,000 g / mol. In some embodiments, the molecular weight of the copolymer is from about 10,000 to about 20,000 g / mol.In some embodiments, the molecular weight of the copolymer is about 20,000 to about 25,000 g / mol. In some embodiments, the molecular weight of the copolymer is about 25,000 to about 30,000 g / mol. In some embodiments, the molecular weight of the copolymer is about 30,000 to about 40,000 g / mol. In some embodiments, the molecular weight of the copolymer is about 2,000 to about 10,000 g / mol. In some embodiments, the molecular weight of the copolymer is about 3,000 to about 7,000 g / mol. In some embodiments, the molecular weight of the copolymer is about 4,000 to about 6,000 g / mol.

[0090] In some embodiments, the molecular weight of the polyacrylamide-based copolymer is from 1,000 g / mol to 35,000 g / mol, from 1,000 g / mol to 30,000 g / mol, from 1,000 g / mol to 25,000 g / mol, from 1,000 g / mol to 20,000 g / mol, from 1,000 g / mol to 15,000 g / mol, from 1,000 g / mol to 10,000 g / mol, from 1,000 g / mol to 7,000 g / mol, from 1,000 g / mol to 6,000 g / mol, from 1,000 g / mol to 5,000 g / mol, from 1,000 g / mol to 4,000 g / mol, from 1,000 g / mol to 3,000 g / mol, 2,000 g / mol to 10,000 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, 3,000 g / mol to 5,000 g / mol, 3,000 g / mol to 4, 000g / mol, 4,000g / mol to 40,000g / mol, 4,000g / mol to 35,000g / mol, 4,000g / mol to 30,000g / mol, 4,000g / mol to 25,000g / mol, 4,000g / mol to 20,000g / mol, 4,000g / mol to 15,000g / mol, 4,000g / mol to 10,000g / mol, 4,000g / mol to 7,000g / mol, 4,000g / mol to 6,000g / mol, 4,000g / mol to 5,000g / mol, 5,000g / mol to 40,000g / mol, 5,000g / mol to 35, 000g / mol, 5,000g / mol to 30,000g / mol, 5,000g / mol to 25,000g / mol, 5,000g / mol to 20,000g / mol, 5,000g / mol to 15,000g / mol, 5,000g / mol to 10,000g / mol, 5,000g / mol to 7,000g / mol, 5,000g / mol to 6,000g / mol, 6,000g / mol to 40,000g / mol, 6,000g / mol to 35,000g / mol, 6,000g / mol to 30,000g / mol, 6,000g / mol to 25,000g / mol, 6,000g / mol to 20,000g / mol, 6,000g / mol to 15,000g / mol, 6,000g / mol to 10,000g / mol, 6,000g / mol to 7,000g / mol, 7,000g / mol to 40,000g / mol, 7,000g / mol to 35,000g / mol, 7,000g / mol to 30,000g / mol, 7,000g / mol to 25,000g / mol, 7,000g / mol to 20,000g / mol, 7, 000g / mol to 15,000g / mol, 7,000g / mol to 10,000g / mol, 10,000g / mol to 40,000g / mol, 10,000g / mol to 35,000g / mol, 10,000g / mol to 30,000g / mol, 10,000g / mol to 25,000g / mol, 10,000g / mol to 20,000g / mol, 10,000g / mol to 15,000g / mol, 15,00 0 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, 20,000 g / mol to 30,000 g / mol, 20,000 The copolymer may have a molecular weight of 1,000 to 40,000 g / mol, such as 25,000 g / mol, 25,000 g / mol, 40,000 g / mol, 35,000 g / mol, 30,000 g / mol, 40,000 g / mol, 30,000 g / mol, 35,000 g / mol, or 40,000 g / mol. In some embodiments, the copolymer has a molecular weight of 1,000 to 30,000 g / mol. In some embodiments, the copolymer has a molecular weight of 10,000 to 20,000 g / mol. In some embodiments, the copolymer has a molecular weight of 15,000 to 20,000 g / mol. In some embodiments, the molecular weight of the copolymer is 20,000 to 25,000 g / mol. In some embodiments, the molecular weight of the copolymer is 25,000 to 30,000 g / mol. In some embodiments, the molecular weight of the copolymer is 30,000 to 40,000 g / mol. In some embodiments, the molecular weight of the copolymer is 2,In some embodiments, the molecular weight of the copolymer is 3,000 to 7,000 g / mol. In some embodiments, the molecular weight of the copolymer is 4,000 to 6,000 g / mol.

[0091] In some embodiments, the polyacrylamide-based copolymer contains a water-soluble carrier monomer having an acrylamide-reactive moiety and a functional dopant monomer (as described herein). In some embodiments, the polyacrylamide-based copolymer contains about 70% to about 98% of the water-soluble carrier monomer having an acrylamide-reactive moiety and about 2% to about 30% of the functional dopant monomer. 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 polyacrylamide-based copolymer contains 70% to 98% of the water-soluble carrier monomer having an acrylamide-reactive moiety and about 2% to about 30% of the functional dopant monomer. 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 non-ionic. In some embodiments, the copolymer is amphiphilic.

[0092] In some embodiments, the polyacrylamide-based copolymer comprises about 70% to about 95% by weight of the water-soluble carrier monomer MORPH and about 5% to about 30% by weight of the functional dopant monomer NIP, the number average molecular weight (Mn) of the copolymer is about 1,000 g / mol to about 10,000 g / mol, and the degree of polymerization is about 10 to about 100. In some embodiments, the polyacrylamide-based copolymer comprises about 74% to about 80% by weight of the water-soluble carrier monomer MORPH and about 20% to about 26% by weight of the functional dopant monomer NIP, the number average molecular weight (Mn) of the copolymer is about 1,000 g / mol to about 5,000 g / mol, and the degree of polymerization is about 10 to about 50. In some embodiments, the polyacrylamide-based copolymer comprises about 77% by weight of the water-soluble carrier monomer MORPH and about 23% by weight of the functional dopant monomer NIP, wherein the number average molecular weight (Mn) of the copolymer is about 3,200 g / mol and the degree of polymerization is about 26.

[0093] In some embodiments, the polyacrylamide-based copolymer comprises 70% to 95% by weight of the water-soluble carrier monomer MORPH and 5% to 30% by weight of the functional dopant monomer NIP, the copolymer having a number average molecular weight (Mn) of 1,000 g / mol to 10,000 g / mol and a degree of polymerization of 10 to 100. In some embodiments, the polyacrylamide-based copolymer comprises 74% to 80% by weight of the water-soluble carrier monomer MORPH and 20% to 26% by weight of the functional dopant monomer NIP, the copolymer having a number average molecular weight (Mn) of 1,000 g / mol to 5,000 g / mol and a degree of polymerization of 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 NIP, and the number average molecular weight (Mn) of the copolymer is 3,200 g / mol and the degree of polymerization is 26.

[0094] In some embodiments, the particles comprise from about 0.01% to about 25% by weight of polyacrylamide-based copolymer, e.g., from about 0.01% to about 20% by weight, from about 0.01% to about 10% by weight, from about 0.01% to about 5% by weight, from about 0.1% to about 5% by weight, from about 0.2% to about 5% by weight, from about 0.3% to about 5% by weight, from about 0.4% to about 5% by weight, from about 0.5% to about 5% by weight, from about 0.6% to about 5% by weight, from about 0.7% to about 5% by weight, from about 0.8% to about 5% by weight, from about 0.9% to about 5% by weight, from about 1% to about 5% by weight, from about 0.1% to about 4% by weight, from about 0.1% to about 3% by weight, from about 0.1% to about 2% by weight, or from about 0.1% to about 1% by weight of polyacrylamide-based copolymer. In some embodiments, the particles comprise about 5% by weight of a polyacrylamide-based copolymer.

[0095] In some embodiments, the particles comprise 0.1% to 25% by weight of polyacrylamide-based copolymer, e.g., 0.1% to 20% by weight, 0.1% to 10% by weight, 0.1% to 5% by weight, 0.2% to 5% by weight, 0.3% to 5% by weight, 0.4% to 5% by weight, 0.5% to 5% by weight, 0.6% to 5% by weight, 0.7% to 5% by weight, 0.8% to 5% by weight, 0.9% to 5% by weight, 1% to 5% by weight, 0.1% to 4% by weight, 0.1% to 3% by weight, 0.1% to 2% by weight, or 0.1% to 1% by weight of polyacrylamide-based copolymer. In some embodiments, the particles comprise 5% to 25% by weight of polyacrylamide-based copolymer, e.g., about 5% to about 10% by weight. In some embodiments, the particles comprise 5% by weight of a polyacrylamide-based copolymer.

[0096] 4.1.3. Optional Particle Components The pharmaceutical compositions of the present disclosure can further comprise one or more additional ingredients, such as excipients. In some embodiments, such additional ingredients or excipients are formulated into particles of the composition.

[0097] The term "excipient" refers to a natural or synthetic substance formulated alongside the active biopharmaceutical agent of a composition, included for purposes of stabilizing or bulking up the biopharmaceutical agent in the final dosage form, and / or to impart therapeutic enhancements, such as facilitating drug absorption, reducing viscosity, increasing or decreasing aqueous or non-aqueous solubility, adjusting tonicity, easing injection site discomfort, lowering the freezing point, or improving stability. Excipients can also be useful in the manufacturing process to aid in handling of the biopharmaceutical agent of interest, such as by promoting powder flow or non-stick properties, in addition to aiding stability, such as preventing denaturation or aggregation, over the expected shelf life.

[0098] The term "pharmaceutically acceptable" ingredient, excipient, or component is one that is suitable for use in humans and / or animals without undue adverse side effects (such as toxicity, irritation, and allergic response) commensurate with a reasonable benefit / risk ratio.

[0099] Additional ingredients or excipients of interest include, for example, various materials for modifying, maintaining, or preserving the pH, osmolality, viscosity, clarity, color, tonicity, odor, sterility, stability, dissolution or release rate, adsorption, or penetration of the composition. Suitable formulation materials include, but are not limited to, amino acids, antimicrobial agents, antioxidants, buffers (e.g., phosphate buffer), chelating agents, complexing agents, sugars (monosaccharides, disaccharides, polysaccharides, other carbohydrates (e.g., mannitol, sorbitol, sucrose, trehalose, lactose, melibiose, cyclodextrin, stachyose, lactosucrose, melezidose, raffinose, inulin, chitosan, alginate, hyaluronate celluloses, dextran, alginate, etc.)), synthetic polymers (e.g., poloxamers, polyvinyl alcohol, polyvinylpyrrolidone, pluronics, etc.), emulsifiers (e.g., polysorbates), salt formation counteractants, preservatives, solvents, sugar alcohols, suspending agents, surfactants or wetting agents, stability enhancers, tonicity enhancers, delivery agents, diluents, other excipients, and / or pharmaceutical adjuvants. Preservatives may be added in accordance with appropriate industry standards. The composition may be formulated as a lyophilizate using appropriate excipient solutions as diluents. Suitable ingredients are non-toxic to recipients at the dosages and concentrations used. Further examples of ingredients that may be used in pharmaceutical formulations are found in Remington's Pharmaceutical Sciences, 16 th Ed.(1980)and 20 th Ed. (2000), Mack Publishing Company, Easton, PA.

[0100] In some embodiments, the composition (eg, particles) further comprises one or more of a stabilizer, a preservative, a filler, a bulking agent, a sugar, a polysaccharide, or a viscosity modifier.

[0101] In certain embodiments, the particles comprise a stabilizer, hi some embodiments, the stabilizer is selected from surfactants, poloxamers, povidone, polyvinylpyrrolidone (PVP) polymers, polyvinyl alcohol (PVA) polymers, polysaccharides (e.g., dextran, alginates), celluloses (e.g., hydroxypropylmethylcellulose (HPMC), methylcellulose (MC)), amphoteric compounds, sugars, salts, and combinations thereof.

[0102] 4.2. Injectable Compositions of Particles Also provided is an injectable pharmaceutical composition comprising the suspension of particles described in the present disclosure in liquid carrier.Described herein are exemplary components of particles that are used in pharmaceutical compositions suitable for injection, and methods for making particles.Also described are components of injectable pharmaceutical compositions other than particles, such as liquid carrier.It is understood that the components of composition can be described in terms of their content in solid particles, and / or in terms of their content in the final injectable pharmaceutical composition, including both the amount of solid particles and the amount of liquid carrier.

[0103] In some embodiments, compositions containing suspended particles of the present disclosure have a copolymer concentration of about 0.01% to about 10% by weight of the composition, such as about 0.01% to about 5% by weight of the composition, in some embodiments, the copolymer concentration is about 0.1% to about 5% by weight of the composition.

[0104] In some embodiments, a composition containing suspended particles of the present disclosure has about 20% by weight or more, e.g., about 25% by weight or more, about 30% by weight or more, about 40% by weight or more, about 50% by weight or more, about 60% by weight or more, about 65% by weight or more, about 70% by weight or more, about 75% by weight or more, about 80% by weight or more, about 85% by weight or more, about 90% by weight or more, or about 95% by weight or more of a biopharmaceutical agent. In some embodiments, the composition comprises between about 20% and 65% by weight of the biopharmaceutical agent, e.g., between about 40% and 65% by weight, or between 35% and 50% by weight of the biopharmaceutical agent.

[0105] In some embodiments, a composition containing suspended particles of the present disclosure has 20% or more by weight of a biopharmaceutical agent, e.g., 25% or more, 30% or more, 40% or more, 50% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 95% or more by weight. In some embodiments, the composition comprises 20% to 80% by weight of a biopharmaceutical agent, e.g., 35% to 80% by weight of a biopharmaceutical agent, such as 40% to 65% by weight, or 35% to 50% by weight.

[0106] In some embodiments, the biopharmaceutical agent is an antibody.

[0107] In some embodiments, the suspension particles contain an antibody. In some embodiments, the antibody is a monoclonal antibody or a fragment thereof. In some embodiments, the composition has an antibody concentration of about 20% by weight or more, e.g., about 25% by weight or more, about 30% by weight or more, about 40% by weight or more, about 50% by weight or more, about 60% by weight or more, about 65% by weight or more, about 70% by weight or more, about 75% by weight or more, about 80% by weight or more, about 85% by weight or more, about 90% by weight or more, or about 95% by weight or more of antibody. In some embodiments, the composition comprises about 20% to 65% by weight of antibody, e.g., about 40% to 65% by weight, or 35% to 50% by weight of antibody.

[0108] In some embodiments, the composition comprises 20% or more by weight of an antibody or fragment thereof, e.g., 25% or more, 30% or more, 40% or more, 50% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 95% or more by weight of the antibody or fragment thereof. In some embodiments, the composition comprises 20% to 80% by weight of the biopharmaceutical agent, e.g., 35% to 80%, 40% to 65%, or 35% to 50% by weight of the antibody.

[0109] In some embodiments, compositions containing suspended particles of the present disclosure have 20% or less by weight of the biopharmaceutical agent, e.g., 15% or less, 10% or less, 9% or less, 8% or less, 7% or less, 6% or less, 5% or less, 4% or less, 3% or less, 2% or less, or 1% or less by weight of the biopharmaceutical agent. In some embodiments, the composition contains 0.1% to 20% by weight of the biopharmaceutical agent, e.g., 0.1% to about 10% by weight, or 0.1% to about 5% by weight. In some embodiments, the composition contains 1% to 20% by weight of the biopharmaceutical agent, e.g., 1% to 10% by weight.

[0110] In some embodiments, the biopharmaceutical agent is a peptide or peptide analog.

[0111] In some embodiments, the suspended particles contain insulin or an analog thereof. In some embodiments, the composition comprises about 1% to about 20%, about 1% to about 15%, about 1% to about 10%, about 1% to about 9%, or about 1% to about 8% insulin or an analog thereof by weight. In some embodiments, the composition comprises about 7%, about 8%, about 9%, or about 10% insulin or an analog thereof by weight. In some embodiments, the composition comprises 1% to 20%, 1% to 15%, 1% to 10%, 1% to 9%, or 1% to 8% insulin or an analog thereof by weight. In some embodiments, the composition comprises 7%, 8%, 9%, or 10% insulin or an analog thereof by weight.

[0112] In some embodiments, the concentration of insulin or insulin analog in the composition is about U50 (i.e., 50 U / mL) to about U1000, about U50 to about U500, about U50 to about U200, about U50 to about U100, about U100 to about U500, or about U100 to about U200. In some embodiments, the concentration of insulin or insulin analog in the composition is about U50, about U100, about U200, about U500, or about U1000. In some embodiments, the composition comprises about 1.7 mg / mL to about 17.5 mg / mL, about 1.7 mg / mL to about 7 mg / mL, about 1.7 mg / mL to about 3.5 mg / mL, about 3.5% by weight to about 17.5 mg / mL, or about 3.5 mg / mL to about 7 mg / mL of insulin or analog thereof. In some embodiments, the composition comprises about 1.7 mg / mL, about 3.5 mg / mL, about 7 mg / mL, or about 17.5 mg / mL of insulin or an analog thereof. In some embodiments, the concentration of insulin or an insulin analog in the composition is between U50 (i.e., 50 U / mL) and U500, between U50 and U200, between U50 and U100, between U100 and U500, or between U100 and U200. In some embodiments, the concentration of insulin or an insulin analog in the composition is between U50, U100, U200, or U500. In some embodiments, the composition comprises between 1.7 mg / mL and 17.5 mg / mL, between 1.7 mg / mL and 7 mg / mL, between 1.7 mg / mL and 3.5 mg / mL, between 3.5 mg / mL and 17.5 mg / mL, or between 3.5 mg / mL and 7 mg / mL of insulin or an analog thereof. In some embodiments, the composition comprises 1.7 mg / mL, 3.5 mg / mL, 7 mg / mL, or 17.5 mg / mL of insulin or an analog thereof.

[0113] In some embodiments, the suspended particles comprise a co-formulation of particles comprising a biopharmaceutical agent. In some embodiments, the suspended particles comprise a co-formulation of a first batch of particles comprising a biopharmaceutical agent that is insulin or an insulin analog, and a second batch of particles comprising a biopharmaceutical agent that is a peptide or peptide analog. In some embodiments, the suspended particles comprise a co-formulation of particles comprising insulin or an insulin analog, and particles comprising pramlintide. In some embodiments, the composition comprises 1.0 mg / mL to 35 mg / mL, 1.0 mg / mL to 17.5 mL, 1.0 mg / mL to 7.0 mg / mL, 1.5 mg / mL to 5.0 mg / mL, 2.0 mg / mL to 5.0 mg / mL, 2.5 mg / mL to 4.5 mg / mL, or 3.0 mg / mL to 4.0 mg / mL of insulin or an analog thereof and 0.2 mg / mL to 2.0 mg / mL, 0.2 mg / mL to 1.5 mg / mL, 0.2 mg / mL to 1.0 mg / mL, 0.2 mg / mL to 1.2 mg / mL, 0.5 mg / mL to 1.0 mg / mL, or 0.6 mg / mL to 0.9 mg / mL of pramlintide. In some embodiments, the composition comprises 1.5 mg / mL, 3.5 mg / mL, 5 mg / mL, 7 mg / mL, 17.5 mg / mL, or 35 mg / mL of insulin or an analog thereof and 0.2, 0.5, 0.6, 0.8, 1.0, or 1.2 mg / mL of pramlintide.

[0114] In some embodiments, the composition comprises a weight to weight ratio of polyacrylamide-based copolymer to biopharmaceutical agent of 1:500 to 1:1.

[0115] In some embodiments, the composition comprises a weight to weight ratio of polyacrylamide-based copolymer to biopharmaceutical agent of 1:10 to 1:2 (e.g., a 1:5 weight to weight ratio).

[0116] In some embodiments, the composition comprises a weight to weight ratio of polyacrylamide-based copolymer to biopharmaceutical agent of 1:25 to 1:15 (e.g., a 1:20 weight to weight ratio).

[0117] 4.2.1. Liquid Carrier As summarized above, the injectable compositions described herein comprise particles suspended in a liquid carrier. In some embodiments, the liquid carrier is non-aqueous. In some embodiments, the liquid carrier is an organic liquid. In some embodiments, the liquid carrier comprises an organic liquid and an aqueous solution. In some embodiments, the liquid carrier is referred to as a pharmaceutically acceptable liquid carrier.

[0118] The term "pharmaceutically acceptable liquid carrier" means a pharmaceutically acceptable solvent, liquid suspending agent, or liquid vehicle for delivering a biopharmaceutical agent of the present disclosure to an animal or human.

[0119] In some embodiments, the organic liquid comprises an organic solvent, an oil, or a combination thereof. In some embodiments, the liquid carrier is an organic solvent. It is understood that, in this disclosure, organic "solvent" does not necessarily dissolve the biopharmaceutical agent and suspended particles of polyacrylamide-based copolymer, but rather is a term used by those skilled in the art to refer to organic liquids of interest that can be utilized in the present injectable pharmaceutical compositions.

[0120] In some embodiments, the liquid carrier comprises an organic solvent and an aqueous solution such as water or a buffer. In some embodiments, the liquid carrier is a mixture of a water-miscible organic liquid that does not dissolve the particles and / or biopharmaceutical agent. In some embodiments, an amount of water is included in the liquid carrier as a viscosity-reducing agent. In some embodiments, the liquid carrier comprises 10% by weight or less of an aqueous solution (e.g., water) in a miscible organic solvent, such that the resulting liquid carrier mixture does not dissolve the particles and / or biopharmaceutical agent.

[0121] In some embodiments, the organic liquid comprises an organic solvent. An organic solvent refers to a low-MW carbon-based substance that can dissolve or disperse one or more other substances. In the pharmaceutical composition of the present disclosure, the organic solvent can be a liquid that can disperse or suspend, but not dissolve, suspended particles (e.g., as described herein). It should be understood that one or more selected excipients can be dissolved in the organic liquid of the composition.

[0122] In some embodiments, the organic liquid is selected from triacylglycerides, diacylglycerides, monoacylglycerides, acetamides, alkyl alcohols, aryl alcohols, aralkyl alcohols, fatty acids or fatty acid esters, oils, alkanes, perfluoroalkanes, propylene glycol monoesters, propylene glycol diesters, butylene glycol monoesters, butylene glycol diesters, polyethylene glycol diesters, and combinations thereof.

[0123] In some embodiments, the organic liquid is a monoacylglyceride. Monoacylglycerides are derived from glycerol and one fatty acid connected to the glycerol via an ester bond. The monoacylglyceride can be 1-monoacylglycerol or 2-monoacylglycerol. In some embodiments, the monoacylglyceride comprises a fatty acid chain (saturated or unsaturated, branched or straight-chain) having 4 to 28 carbon atoms, such as 8 to 20 carbon atoms. In some embodiments, the fatty acid chain has 8 to 12 carbon atoms, or 8 to 10 carbon atoms. In some embodiments, the fatty acid ester group of the propylene glycol diester is selected from oleic acid, myristic acid, caprylic acid, and capric acid. In some embodiments, the fatty acid is a saturated fatty acid. In some embodiments, the fatty acid is an unsaturated fatty acid, such as linoleic acid, myristoleic acid, palmitoleic acid, or oleic acid. In some embodiments, the fatty acids are selected from hexanoic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, myristoleic acid, palmitoleic acid, and oleic acid.

[0124] In some embodiments, the organic liquid is a diacylglyceride. Diacylglycerides are derived from glycerol and two fatty acids connected to glycerol via an ester bond. The diacylglyceride can be 1,2-diacylglycerol or 1,3-diacylglycerol. In some embodiments, the diacylglyceride comprises two fatty acid chains (saturated or unsaturated, branched or straight chain), each independently having 4 to 28 carbon atoms, such as 8 to 20 carbon atoms. In some embodiments, the fatty acid chains each have 8 to 12 carbon atoms or 8 to 10 carbon atoms. In some embodiments, the fatty acid ester group of the propylene glycol diester is selected from oleic acid, myristic acid, caprylic acid, capric acid, or any combination thereof. In some embodiments, the fatty acid is a saturated fatty acid. In some embodiments, the fatty acid is an unsaturated fatty acid, such as linoleic acid, myristoleic acid, palmitoleic acid, or oleic acid. In some embodiments, the fatty acid comprises both saturated and unsaturated fatty acids. In some embodiments, the fatty acids are selected from hexanoic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, myristoleic acid, palmitoleic acid, and oleic acid.

[0125] In some embodiments, the organic liquid comprises a triacylglyceride. In some embodiments, the triacylglyceride is triacetin.

[0126] In some embodiments, the organic liquid comprises a triacylglyceride having three fatty acid chains. In some embodiments, the organic liquid comprises a triacylglyceride having two fatty acid chains. In some embodiments, the organic liquid comprises a triacylglyceride having one fatty acid chain. In some embodiments, the fatty acid chains are each independently a hydrocarbon chain (saturated or unsaturated, branched or straight-chain) containing 4 to 28 carbon atoms, such as 8 to 20, 8 to 12, or 8 to 10 carbon atoms. In some embodiments, the fatty acid chain is selected from caprylic acid and capric acid. In some embodiments, the fatty acid is a saturated fatty acid. In some embodiments, the fatty acid is an unsaturated fatty acid, such as linoleic acid, myristoleic acid, palmitoleic acid, or oleic acid. In some embodiments, the fatty acid comprises both saturated and unsaturated fatty acids. In some embodiments, the fatty acid is selected from hexanoic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, myristoleic acid, palmitoleic acid, and oleic acid. Triacylglyceride liquids of interest include, but are not limited to, Miglyol® 829.

[0127] In some embodiments, the organic liquid comprises acetamide. Acetamide liquids of interest include, but are not limited to, acetamide (CH3CONH2), N-alkyl-acetamides, and N,N-dialkyl-acetamides. In some embodiments, the acetamide is selected from acetamide (CH3CONH2), N-methylacetamide, N-ethylacetamide, N,N-diethylacetamide, and N,N-dimethylacetamide. In some embodiments, the acetamide is N,N-dimethylacetamide (DMAc).

[0128] In some embodiments, the organic liquid comprises an alkyl alcohol, an aryl alcohol, an aralkyl alcohol, or a combination thereof. In some embodiments, the organic liquid comprises octanol. In some embodiments, the organic liquid comprises isopropyl alcohol. In some embodiments, the organic liquid comprises ethyl alcohol. In some embodiments, the organic liquid comprises benzyl alcohol.

[0129] In some embodiments, the organic liquid comprises an alkyl benzoate, an aryl benzoate, an aralkyl benzoate, or any combination thereof, hi some embodiments, the organic liquid comprises benzyl benzoate.

[0130] In some embodiments, the organic liquid comprises a polar aprotic solvent, hi some embodiments, the polar aprotic solvent comprises dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), or a mixture thereof.

[0131] In some embodiments, the organic liquid comprises a propylene glycol (PG) solvent.

[0132] In some embodiments, the organic liquid comprises a propylene glycol diester. In some embodiments, the propylene glycol diester is a propylene glycol diester of a fatty acid(s). In some embodiments, the fatty acid ester group of the propylene glycol diester comprises a hydrocarbon chain (saturated or unsaturated, branched or straight chain) containing 4 to 28 carbon atoms, such as 8 to 20 carbon atoms. In some embodiments, the fatty acid ester group of the propylene glycol diester is selected from oleic acid, myristic acid, caprylic acid, capric acid, or any combination thereof.

[0133] In some embodiments, the propylene glycol diester comprises a propylene glycol diester of caprylic acid, a propylene glycol diester of capric acid, or a combination thereof. In some embodiments, the propylene glycol diester comprises a diester of caprylic acid and / or capric acid. In some embodiments, the propylene glycol diester is propylene glycol dicaprylate. In some embodiments, the propylene glycol diester is propylene glycol dicaprate. Propylene glycol diesters of interest include, but are not limited to, Miglyol® 840.

[0134] In some embodiments, the organic liquid is a propylene glycol monoester. In some embodiments, the propylene glycol monoester comprises one fatty acid ester having a chain (saturated or unsaturated, branched or straight) of 4 to 28 carbon atoms, such as 8 to 20 carbon atoms. In some embodiments, the fatty acid chain has 8 to 10 carbon atoms. In some embodiments, the fatty acid ester group of the propylene glycol monoester is selected from oleic acid, myristic acid, caprylic acid, and capric acid. In some embodiments, the fatty acid is a saturated fatty acid. In some embodiments, the fatty acid is an unsaturated fatty acid, such as linoleic acid, myristoleic acid, palmitoleic acid, or oleic acid. In some embodiments, the fatty acid is selected from hexanoic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, myristoleic acid, palmitoleic acid, and oleic acid.

[0135] In some embodiments, the organic liquid comprises a butylene glycol diester. In some embodiments, the butylene glycol diester is a butylene glycol diester of a fatty acid(s). In some embodiments, the fatty acid ester group of the butylene glycol diester comprises a hydrocarbon chain (saturated or unsaturated, branched or straight-chain) containing 4 to 28 carbon atoms, such as 8 to 20 carbon atoms, 8 to 12 carbon atoms, or 8 to 10 carbon atoms. In some embodiments, the fatty acid ester group of the butylene glycol diester is selected from oleic acid, myristic acid, caprylic acid, capric acid, or any combination thereof. In some embodiments, the butylene glycol solvent comprises a propylene glycol diester of caprylic acid, a propylene glycol diester of capric acid, or a combination thereof. In some embodiments, the butylene glycol diester comprises a diester of caprylic acid and capric acid (e.g., a butylene glycol ester of caprylic / capric acid). In some embodiments, the fatty acid is a saturated fatty acid. In some embodiments, the fatty acid is an unsaturated fatty acid such as linoleic acid, myristoleic acid, palmitoleic acid, or oleic acid. In some embodiments, the fatty acid comprises both saturated and unsaturated fatty acids. In some embodiments, the fatty acid is selected from hexanoic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, myristoleic acid, palmitoleic acid, and oleic acid. Butylene glycol diesters of interest include, but are not limited to, Miglyol® 8810.

[0136] In some embodiments, the organic liquid is a butylene glycol monoester. In some embodiments, the butylene glycol monoester comprises one fatty acid ester having a chain (saturated or unsaturated, branched or straight) of 4 to 28 carbon atoms, such as 8 to 20 carbon atoms. In some embodiments, the fatty acid chain has 8 to 10 carbon atoms. In some embodiments, the fatty acid ester group of the butylene glycol monoester is selected from oleic acid, myristic acid, caprylic acid, and capric acid. In some embodiments, the fatty acid is a saturated fatty acid. In some embodiments, the fatty acid is an unsaturated fatty acid, such as linoleic acid, myristoleic acid, palmitoleic acid, or oleic acid. In some embodiments, the fatty acid is selected from hexanoic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, myristoleic acid, palmitoleic acid, and oleic acid.

[0137] In some embodiments, the organic liquid comprises a polyethylene glycol (PEG) solvent. In some embodiments, the PEG solvent is PEG200, PEG400, or a combination thereof.

[0138] In some embodiments, the organic liquid is a polyethylene glycol diester. In some embodiments, the polyethylene glycol portion of the polyethylene glycol diester has an average MW of 500 or less, such as 400 or less, 200-400, or 300-400. In some embodiments, the polyethylene glycol diester has two fatty acid ester groups attached to a linear polyethylene glycol. In some embodiments, the polyethylene glycol diester comprises a fatty acid ester having a chain (saturated or unsaturated, branched or linear) of 4 to 28 carbon atoms, such as 8 to 20 carbon atoms. In some embodiments, the fatty acid chain has 8 to 10 carbon atoms. In some embodiments, the fatty acid ester group of the polyethylene glycol diester is selected from oleic acid, myristic acid, caprylic acid, and capric acid. In some embodiments, the fatty acid is a saturated fatty acid. In some embodiments, the fatty acid is an unsaturated fatty acid, such as linoleic acid, myristoleic acid, palmitoleic acid, or oleic acid. In some embodiments, the fatty acid comprises both saturated and unsaturated fatty acids. In some embodiments, the fatty acids are selected from hexanoic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, myristoleic acid, palmitoleic acid, and oleic acid.

[0139] In some embodiments, the organic liquid comprises a fatty acid or fatty acid ester. In some embodiments, the fatty acid or fatty acid ester comprises a hydrocarbon chain (saturated or unsaturated, branched or straight-chain) containing 4 to 28 carbon atoms, such as 8 to 20 carbon atoms, 8 to 16 carbon atoms, 8 to 12 carbon atoms, or 8 to 10 carbon atoms. In some embodiments, the fatty acid is a saturated fatty acid. In some embodiments, the fatty acid is an unsaturated fatty acid, such as linoleic acid, myristoleic acid, palmitoleic acid, or oleic acid. In some embodiments, the fatty acid is selected from hexanoic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitoleic acid, and oleic acid. In some embodiments, the fatty acid is oleic acid, myristic acid, caprylic acid, capric acid, or any combination thereof. In some embodiments, the fatty acid ester is derived from oleic acid, myristic acid, caprylic acid, or capric acid. In some embodiments, the fatty acid ester is derived from hexanoic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, myristoleic acid, palmitoleic acid, or oleic acid. In some embodiments, the fatty acid ester is a (C1-C6)-alkyl or substituted (C1-C6)-alkyl ester of a fatty acid (e.g., as described herein). In some embodiments, the fatty acid ester comprises ethyl oleate, isopropyl myristate, or a combination thereof.

[0140] In some embodiments, the organic liquid comprises an oil (e.g., a vegetable oil or an animal oil). In some embodiments, the oil comprises coconut oil, cottonseed oil, fish oil, grapeseed oil, hazelnut oil, hydrogenated vegetable oil, lime oil, olive oil, palm seed oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, walnut oil, or any combination thereof. In some embodiments, the oil comprises sesame oil. In some embodiments, the oil comprises safflower oil.

[0141] In some embodiments, the organic solvent comprises an alkane or a perfluoroalkane, hi some embodiments, the organic solvent comprises perfluorohexyl octane, perfluorooctane, octane, perfluorodecalin, perfluorobutyl pentane, tetradecane, and any combination thereof.

[0142] In some embodiments, the organic solvent comprises ethyl lactate, ethyl acetate, propyl acetate, or any combination thereof.

[0143] Organic liquids of interest include, but are not limited to, benzyl benzoate, ethyl oleate, triacetin, dimethylacetamide (DMAc), Miglyol 840, Miglyol 829, Miglyol 8810, Miglyol 812, Miglyol 812, and the like.

[0144] In some embodiments, the organic liquid comprises a blend of organic liquids (e.g., as described herein), e.g., a blend of organic solvents. In some embodiments, the organic liquid is a blend comprising an organic selected from triacylglycerides, diacylglycerides, monoacylglycerides, acetamides, alkyl alcohols, aryl alcohols, aralkyl alcohols, fatty acids or fatty acid esters, oils, alkanes, perfluoroalkanes, propylene glycol monoesters, propylene glycol diesters, butylene glycol monoesters, butylene glycol diesters, and polyethylene glycol diesters. In some embodiments, the organic liquid blend comprises an additional organic solvent. In some embodiments, the organic liquid is a blend of two or more organic liquids independently selected from triacylglycerides, diacylglycerides, monoacylglycerides, acetamides, alkyl alcohols, aryl alcohols, aralkyl alcohols, fatty acids or fatty acid esters, oils, alkanes, perfluoroalkanes, propylene glycol monoesters, propylene glycol diesters, butylene glycol monoesters, butylene glycol diesters, and polyethylene glycol diesters.

[0145] In some embodiments, the organic liquid blend has a lower viscosity than one or more of the individual liquids or solvents in the blend alone. In some embodiments, the viscosity of the organic liquid blend is 25 cp or less at 25° C., such as 23 cp or less, 20 cp or less, 18 cp or less, 15 cp or less, 12 cp or less, 10 cp or less, 8 cp or less, 5 cp or less, or even less. In some embodiments, the organic liquid blend comprises a blend of triacylglycerides and one or more organic solvents having a lower viscosity than the triacylglycerides. In some embodiments, the triacylglyceride is triacetin. In some embodiments, the organic liquid blend comprises a blend of propylene glycol and one or more organic solvents having a lower viscosity than propylene glycol. In some embodiments, the propylene glycol is a propylene glycol diester or propylene glycol monoester. In some embodiments, the propylene glycol diester or monoester comprises an ester derived from oleic acid, myristic acid, caprylic acid, capric acid, or any combination thereof. In some embodiments, the ester is a (C4-C6) ester of a fatty acid. 20 )-alkyl or substituted (C4-C 20 )-alkyl esters. In some embodiments, the propylene glycol diesters or monoesters include diesters of caprylic and capric acid (e.g., propylene glycol, dicaprylate, dicaprate).

[0146] In some embodiments, the organic liquid blend comprises 50-90% v / v triacetin and 10-50% v / v of one or more organic solvents having a lower viscosity than triacetin. In some embodiments, the organic liquid blend comprises 50-90% v / v triacetin, such as 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or 90% v / v triacetin. In some embodiments, the organic liquid blend comprises 10%-50% of one or more organic solvents having a lower viscosity than triacetin, such as 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% v / v of one or more organic solvents having a lower viscosity than triacetin. In some embodiments, the organic liquid blend comprises 50% v / v triacetin and 50% v / v of one or more organic solvents having a lower viscosity than triacetin. In some embodiments, the organic liquid blend comprises 55% v / v triacetin and 45% v / v of one or more organic solvents having a lower viscosity than triacetin. In some embodiments, the organic liquid blend comprises 60% v / v triacetin and 40% v / v of one or more organic solvents having a lower viscosity than triacetin. In some embodiments, the organic liquid blend comprises 65% v / v triacetin and 35% v / v of one or more organic solvents having a lower viscosity than triacetin. In some embodiments, the organic liquid blend comprises 70% v / v triacetin and 30% v / v of one or more organic solvents having a lower viscosity than triacetin. In some embodiments, the organic liquid blend comprises 75% v / v triacetin and 25% v / v of one or more organic solvents having a lower viscosity than triacetin. In some embodiments, the organic liquid blend comprises 80% v / v triacetin and 20% v / v of one or more organic solvents having a lower viscosity than triacetin. In some embodiments, the organic liquid blend comprises 85% v / v triacetin and 15% v / v of one or more organic solvents having a lower viscosity than triacetin, hi some embodiments, the organic liquid blend comprises 90% v / v triacetin and 10% v / v of one or more organic solvents having a lower viscosity than triacetin.

[0147] In some embodiments, the organic liquid blend comprises 50-90% v / v propylene glycol and 10-50% v / v of one or more organic solvents having a lower viscosity than propylene glycol. In some embodiments, the organic liquid blend comprises 50-90% v / v propylene glycol, such as 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or 90% v / v propylene glycol. In some embodiments, the organic liquid blend comprises 10-50% v / v of one or more organic solvents having a lower viscosity than triacetin, such as 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% v / v of one or more organic solvents having a lower viscosity than propylene glycol. In some embodiments, the organic liquid blend comprises 50% v / v propylene glycol and 50% v / v of one or more organic solvents having a lower viscosity than propylene glycol. In some embodiments, the organic liquid blend comprises 55% v / v propylene glycol and 45% v / v of one or more organic solvents with a lower viscosity than propylene glycol. In some embodiments, the organic liquid blend comprises 60% v / v propylene glycol and 40% v / v of one or more organic solvents with a lower viscosity than propylene glycol. In some embodiments, the organic liquid blend comprises 65% v / v propylene glycol and 35% v / v of one or more organic solvents with a lower viscosity than propylene glycol. In some embodiments, the organic liquid blend comprises 70% v / v propylene glycol and 30% v / v of one or more organic solvents with a lower viscosity than propylene glycol. In some embodiments, the organic liquid blend comprises 75% v / v propylene glycol and 25% v / v of one or more organic solvents with a lower viscosity than propylene glycol. In some embodiments, the organic liquid blend comprises 80% v / v propylene glycol and 20% v / v of one or more organic solvents with a lower viscosity than propylene glycol. In some embodiments, the organic liquid blend comprises 85% v / v propylene glycol and 15% v / v of one or more organic solvents that have a lower viscosity than propylene glycol.In some embodiments, the organic liquid blend comprises 90% v / v propylene glycol and 10% v / v of one or more organic solvents that have a lower viscosity than propylene glycol.

[0148] In some embodiments, the organic liquid blend comprises triacetin and an organic solvent having a lower viscosity than triacetin. In some embodiments, the organic liquid comprises triacetin and two or more organic solvents having a lower viscosity than triacetin. In some embodiments, the one or more solvents having a lower viscosity than triacetin are selected from DMAc, benzyl benzoate, benzyl alcohol, ethanol, isopropyl alcohol, ethyl lactate, perfluorohexyl octane, perfluorooctane, octane, perfluorodecalin, perfluorobutyl pentane, methoxyflurane, octanol, ethyl acetate, propyl acetate, ethyl oleate, and isopropyl myristate. In some embodiments, the one or more solvents having a lower viscosity than triacetin are selected from DMAc, benzyl benzoate, and benzyl alcohol. In some embodiments, the solvent having a lower viscosity than triacetin is DMAc. In some embodiments, the solvent having a lower viscosity than triacetin is benzyl benzoate. In some embodiments, the solvent having a lower viscosity than triacetin is benzyl alcohol. In some embodiments, the solvent having a lower viscosity than triacetin comprises a combination of DMAc and benzyl alcohol. Thus, in some embodiments, the organic liquid comprises a blend of triacetin, DMAc, and benzyl alcohol. In some embodiments, the solvent having a lower viscosity than triacetin comprises a v / v ratio of DMAc to benzyl alcohol of 2:1 to 3:1. In some embodiments, the solvent having a lower viscosity than triacetin comprises a combination of DMAc and benzyl benzoate. In some embodiments, the solvent having a lower viscosity than triacetin comprises a v / v ratio of DMAc to benzyl benzoate of 2:1 to 3:1. In some embodiments, the benzyl alcohol is present in an amount of 10% v / v or less of the total volume of the organic liquid.

[0149] In some embodiments, the organic liquid comprises a blend of triacylglyceride and acetamide. In some embodiments, the triacylglyceride is triacetin. In some embodiments, the acetamide is DMAc. In some embodiments, the organic liquid comprises a volume-to-volume (v / v) ratio of triacetin to DMAc of 1:1 to 9:1, such as a 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, 5:1, 5.5:1, 6:1, 6.5:1, 7:1, 7.5:1, 8:1, 8.5:1, or 9:1 ratio of triacetin to DMAc. In some embodiments, the organic liquid comprises a v / v ratio of triacetin to DMAc of 1:1 to 3:1, such as a v / v ratio of 1:1, 1.2:1, 1.4:1, 1.6:1, 1.8:1, 2:1, 2.2:1, 2.4:1, 2.6:1, 2.8:1, or 3:1. In some embodiments, the organic liquid comprises a v / v ratio of triacetin to DMAc of 3:1 to 5:1, such as a v / v ratio of 3:1, 3.2:1, 3.4:1, 3.6:1, 3.8:1, 4:1, 4.2:1, 4.4:1, 4.6:1, 4.8:1, or 5:1. In some embodiments, the organic liquid comprises a v / v ratio of triacetin to DMAc of 5:1 to 7:1, such as a v / v ratio of 5:1, 5.2:1, 5.4:1, 5.6:1, 5.8:1, 6:1, 6.2:1, 6.4:1, 6.6:1, 6.8:1, or 7:1. In some embodiments, the organic liquid comprises a v / v ratio of triacetin to DMAc of 7:1 to 9:1, such as a v / v ratio of 7:1, 7.2:1, 7.4:1, 7.6:1, 7.8:1, 8:1, 8.2:1, 8.4:1, 8.6:1, 8.8:1, or 9:1.

[0150] In some embodiments, the organic liquid comprises a 1:1 v / v ratio of triacetin to DMAc, in some embodiments, a 3:1 v / v ratio of triacetin to DMAc, in some embodiments, a 9:1 v / v ratio of triacetin to DMAc.

[0151] In some embodiments, the organic liquid comprises a blend of a triacylglyceride and an aralkyl alcohol. In some embodiments, the triacylglyceride is triacetin. In some embodiments, the aralkyl alcohol is benzyl alcohol. In some embodiments, the organic liquid comprises a volume-to-volume (v / v) ratio of triacetin to benzyl alcohol of 1:1 to 9:1, such as a v / v ratio of 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, 5:1, 5.5:1, 6:1, 6.5:1, 7:1, 7.5:1, 8:1, 8.5:1, or 9:1. In some embodiments, the organic liquid comprises a v / v ratio of triacetin to benzyl alcohol of 1:1 to 3:1, such as a v / v ratio of 1:1, 1.2:1, 1.4:1, 1.6:1, 1.8:1, 2:1, 2.2:1, 2.4:1, 2.6:1, 2.8:1, or 3:1. In some embodiments, the organic liquid comprises a v / v ratio of triacetin to benzyl alcohol of 3:1 to 5:1, such as a v / v ratio of 3:1, 3.2:1, 3.4:1, 3.6:1, 3.8:1, 4:1, 4.2:1, 4.4:1, 4.6:1, 4.8:1, or 5:1. In some embodiments, the organic liquid comprises a v / v ratio of triacetin to benzyl alcohol of 5:1 to 7:1, such as a v / v ratio of 5:1, 5.2:1, 5.4:1, 5.6:1, 5.8:1, 6:1, 6.2:1, 6.4:1, 6.6:1, 6.8:1, or 7:1. In some embodiments, the organic liquid comprises a v / v ratio of triacetin to benzyl alcohol of 7:1 to 9:1, such as a v / v ratio of 7:1, 7.2:1, 7.4:1, 7.6:1, 7.8:1, 8:1, 8.2:1, 8.4:1, 8.6:1, 8.8:1, or 9:1.

[0152] In some embodiments, the organic liquid comprises a blend of a triacylglyceride and an aralkyl benzoate. In some embodiments, the triacylglyceride is triacetin. In some embodiments, the aralkyl benzoate is benzyl benzoate. In some embodiments, the organic liquid comprises a volume-to-volume (v / v) ratio of triacetin to benzyl benzoate of 1:1 to 9:1, such as a v / v ratio of 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, 5:1, 5.5:1, 6:1, 6.5:1, 7:1, 7.5:1, 8:1, 8.5:1, or 9:1. In some embodiments, the organic liquid comprises a v / v ratio of triacetin to benzyl benzoate of 1:1 to 3:1, such as a v / v ratio of 1:1, 1.2:1, 1.4:1, 1.6:1, 1.8:1, 2:1, 2.2:1, 2.4:1, 2.6:1, 2.8:1, or 3:1. In some embodiments, the organic liquid comprises a v / v ratio of triacetin to benzyl benzoate of 3:1 to 5:1, such as a v / v ratio of 3:1, 3.2:1, 3.4:1, 3.6:1, 3.8:1, 4:1, 4.2:1, 4.4:1, 4.6:1, 4.8:1, or 5:1. In some embodiments, the organic liquid comprises a v / v ratio of triacetin to benzyl benzoate of 5:1 to 7:1, such as a v / v ratio of 5:1, 5.2:1, 5.4:1, 5.6:1, 5.8:1, 6:1, 6.2:1, 6.4:1, 6.6:1, 6.8:1, or 7:1. In some embodiments, the organic liquid comprises a v / v ratio of triacetin to benzyl benzoate of 7:1 to 9:1, such as a v / v ratio of 7:1, 7.2:1, 7.4:1, 7.6:1, 7.8:1, 8:1, 8.2:1, 8.4:1, 8.6:1, 8.8:1, or 9:1.

[0153] In some embodiments, the composition comprises a weight to weight ratio of liquid carrier to particles of 1:5 to 4:1. In some embodiments, the composition comprises a weight to weight ratio of liquid carrier to particles of 1:1 to 2:1. In some embodiments, the composition comprises a weight to weight ratio of liquid carrier to particles of 1:1 to 1.5:1.

[0154] In some embodiments, the composition comprises 200 mg / mL to 850 mg / mL of particles in a liquid carrier. In some embodiments, the composition comprises 300 mg / mL to 500 mg / mL of particles in a liquid carrier. In some embodiments, the composition comprises 400 mg / mL to 600 mg / mL of particles, such as 450 mg / mL to 550 mg / mL of particles in a liquid carrier.

[0155] 4.3. Loaded Syringe The present disclosure also provides a syringe loaded with an injectable pharmaceutical composition (e.g., as described herein), e.g., a syringe pre-loaded with an injectable pharmaceutical composition described herein prior to expulsion of the composition from the syringe.

[0156] In some embodiments, the loaded syringe is configured to dispense the composition at a flow rate of 0.1 mL / min or greater, such as 0.5 mL / min or greater, 1 mL / min or greater, 2 mL / min or greater, 5 mL / min or greater, 10 mL / min or greater, 15 mL / min or greater, or 20 mL / min or greater, in response to a force applied to the syringe of 70 N or less. In some embodiments, the syringe is configured to dispense the composition at a flow rate of 0.1 mL / min or greater, such as 0.5 mL / min or greater, 1 mL / min or greater, 2 mL / min or greater, 5 mL / min or greater, 10 mL / min or greater, 15 mL / min or greater, or 20 mL / min or greater, in response to a force applied to the syringe of 50 N or less.

[0157] In some embodiments, the loaded syringe has a needle having a size of 18 to 32 gauge (G). In some embodiments, the needle is an 18G needle. In some embodiments, the needle is a 21G needle. In some embodiments, the needle is a 22G needle. In some embodiments, the needle is a 24G needle. In some embodiments, the needle is a 25G needle. In some embodiments, the needle is a 26G needle. In some embodiments, the needle is a 27G needle. In some embodiments, the needle is a 30G needle. In some embodiments, the needle is a 32G needle.

[0158] In some embodiments of any of the needles described herein, the needle is an ultra-thin wall needle (UTW). It is understood that a UTW needle can have a larger inner diameter than a conventional gauge needle.

[0159] 4.4.Administration Method Also provided in this disclosure are methods of administering a biopharmaceutical to a subject by injection. In some embodiments, the method comprises injecting a composition (e.g., as described herein) into a subject in need thereof, thereby administering a therapeutically effective dose of the biopharmaceutical agent to the subject.

[0160] In some embodiments of the method, the injecting is performed using a loaded syringe, such as a pre-loaded syringe, containing the composition.

[0161] 4.5. Preparation method As summarized herein, methods of preparing an injectable pharmaceutical composition (e.g., as described herein) are also provided. In some embodiments, the method of preparing an injectable pharmaceutical composition comprises: a) a biopharmaceutical agent; providing a mixture comprising: a polyacrylamide-based copolymer in water; b) removing water from the mixture to obtain particles comprising the biopharmaceutical agent and the polyacrylamide-based copolymer; c) contacting a liquid carrier with the particles to form a suspension of the particles in the organic liquid.

[0162] In some embodiments of the method, in step b), water is removed via spray drying. In some embodiments of the method, in step b), water is removed via freeze drying. In some embodiments of the method, in step b), water is removed via electrospray drying.

[0163] In some embodiments, the method further comprises a milling step after lyophilization but before step c).

[0164] In some embodiments, the particles obtained in step b) have an average diameter as described herein, for example, less than or equal to 100 microns.

[0165] In some embodiments, step c) of the method comprises sonicating the particles in the liquid carrier.

[0166] 4.6.Definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Methods and materials for use in this application are described herein; other suitable methods and materials known in the art are also used in some embodiments of this disclosure. The materials, methods, and examples are merely illustrative and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. When trade names are used herein, they include the product formulations, over-the-counter drugs, and active pharmaceutical ingredient(s) of the trade name product unless the context dictates otherwise.

[0167] The terms "subject" and "patient" are used interchangeably. A subject can be a mammal, e.g., a human, such as a non-primate (e.g., cows, pigs, horses, cats, dogs, goats, rabbits, rats, mice, etc.) or a primate (e.g., monkeys, apes, and humans). In certain embodiments, the subject is a mammal, e.g., a human, diagnosed with a disease or disorder provided herein. In another embodiment, the subject is a mammal, e.g., a human, at risk of developing a disease or disorder provided herein. In a specific embodiment, the subject is a human.

[0168] The terms "therapies" and "therapy" are used in their broadest sense as understood in the clinical field.

[0169] In this disclosure, the terms "a," "an," and "the" are used to include one or more than one unless the context clearly dictates otherwise. The term "or" is used to refer to a non-exclusive "or" unless otherwise indicated. The phrase "at least one of A and B" has the same meaning as "A, B, or A and B." Additionally, it should be understood that terms or terminology used in this disclosure, unless otherwise defined, are for descriptive purposes only and not limiting. The use of section headings is intended to aid in the reading of the document and is not to be construed as limiting. Information associated with a section heading may be found within or outside of that particular section.

[0170] Values ​​expressed in range format should be interpreted flexibly to include not only the numerical values ​​explicitly recited as the limits of the range, but also all individual numerical values ​​or subranges subsumed within that range, as if each numerical value and subrange were explicitly recited. For example, a range of "about 0.1% to about 5%" or "about 0.1% to 5%" should be interpreted to include not 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 recited range. Unless otherwise indicated, the phrase "about X to Y" has the same meaning as "about X to about Y." Similarly, unless otherwise indicated, the phrase "about X, Y, or about Z" has the same meaning as "about X, about Y, or about Z."

[0171] The term "about" as used in this disclosure allows for a degree of variation of a stated value or range that is within 5% of the stated limits of the stated value or range.

[0172] In the methods described herein, acts may be performed in any order unless a temporal or operational sequence is explicitly recited. Furthermore, specified acts may be performed simultaneously unless explicit claim language recites them being performed separately. For example, a claimed act of doing X and a claimed act of doing Y may be performed simultaneously in a single operation, and the resulting process would fall within the literal scope of the claimed process.

[0173] The term "polymer" refers to a substance or material made up of repeating monomeric subunits.

[0174] As used herein, "acrylamide monomer" refers to a monomeric species having an acrylamide functional group. The term "acrylamide monomer" includes not only monomeric acrylamide but also derivatives of monomeric acrylamide. Examples of acrylamide monomers include, but are not limited to, acrylamide (AM), N-(3-methoxypropionyl)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).

[0175] The term "polyacrylamide-based copolymer" refers to a polymer formed from the polymerization of two or more monomeric species, at least one of which has an acrylamide functional group (acrylamide monomer), and in which the monomers are structurally distinct. In some embodiments, a polyacrylamide-based copolymer is formed from the polymerization of two structurally distinct acrylamide monomers (two structurally distinct monomers, each with an acrylamide functional group). The resulting copolymer can be an alternating copolymer, in which the monomeric species are linked alternately; a random copolymer, in which the monomeric species are linked to each other within the polymer chain without a defined pattern; a block copolymer, in which polymer blocks of one monomeric species are linked to polymer blocks composed of another monomeric species; or a graft copolymer, in which the main polymer chain is composed of one monomeric species and polymer blocks of another monomeric species are linked to the main polymer chain as side branches. In some embodiments, the polyacrylamide-based copolymers of the present disclosure are formed from the polymerization of a water-soluble carrier monomer and a functional dopant monomer. In some embodiments, the polyacrylamide-based copolymers of the present disclosure are random copolymers.

[0176] As defined herein, the term "water-soluble carrier monomer" refers to an acrylamide monomer species that is a water-soluble species within a polyacrylamide-based copolymer. In some embodiments, the water-soluble carrier monomer is the predominant species within the polyacrylamide-based copolymer. In some embodiments, the water-soluble carrier monomer confers water solubility to the copolymer. In some embodiments, the water-soluble carrier monomer within the polyacrylamide-based copolymer provides an inert barrier at an interface of an aqueous formulation to prevent protein-protein interactions. In some embodiments, the interface is an air-water interface. In some embodiments, the interface is an inclusion-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 tubing. In some embodiments, the interface is an interface between a liquid and a catheter. In some embodiments, the inclusion-water interface is within a pump system. In some embodiments, the inclusion-water interface is within 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-methoxypropoyl)acrylamide (MPAM), 4-acryloylmorpholine (MORPH), N,N-dimethylacrylamide (DMA), and N-hydroxyethylacrylamide (HEAM).

[0177] As used herein, the term "functional dopant monomer" refers to an acrylamide monomer species that has physicochemical properties (e.g., hydrophobicity, charge) that differ from those of the water-soluble carrier monomer. In some embodiments, functional dopant monomers in polyacrylamide-based copolymers promote the association of the polymer to interfaces, which 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 function as stabilizing moieties to promote interactions with biomolecules, such as proteins, peptides, antibodies, antibody-drug conjugates, nucleic acids, lipid particles, and combinations thereof (e.g., to prevent biomolecular aggregation). Functional dopant monomers can be further classified based on their chemical composition into hydrogen-bonding monomers, ionic monomers, hydrophobic monomers, and aromatic monomers. Typically, the functional dopant monomer is copolymerized at a lower weight percentage compared to the water-soluble carrier monomer.

[0178] The term "polymerization" refers to a process in which monomer molecules undergo a chemical reaction 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.

[0179] The term "degree of polymerization" (DP) refers to the number of monomer units in a polymer. It is calculated by dividing the average molecular weight of a polymer sample by the molecular weight of the monomer. As defined herein, the average molecular weight of a polymer can be represented by the number average molecular weight (Mn), weight average molecular weight (Mw), Z-average molecular weight (Mz), or the molecular weight at the peak maximum of the molecular weight distribution curve (Mp). The average molecular weight of a polymer can be determined by various analytical characterization techniques known to those skilled in the art, such as size exclusion chromatography (SEC), static light scattering (SLS) analysis, multi-angle laser light scattering (MALLS) analysis, nuclear magnetic resonance spectroscopy (NMR), intrinsic viscosity measurement (IV), melt flow index (MFI), and matrix-assisted laser desorption / ionization mass spectrometry (MALDI-MS), and combinations thereof. The degree of polymerization can also be determined experimentally using suitable analytical methods known in the art, such as nuclear magnetic spectroscopy (NMR), Fourier transform infrared spectroscopy (FT-IR), and Raman spectroscopy.

[0180] The term "amphiphilic" refers to a chemical substance that has both hydrophilic (water-loving, polar) and lipophilic or hydrophobic (lipid-loving, non-polar) properties. Examples of common amphiphilic compounds include detergents, soaps, surfactants, lipoproteins, and phospholipids. In some embodiments, the amphiphile is a charged species. In some embodiments, the amphiphile is a neutral species. In some embodiments, the copolymers incorporated into the particles of the present disclosure are amphiphilic because they contain both hydrophilic comonomers and lipophilic or hydrophobic comonomers.

[0181] As used herein, " lipid-based vehicle " refers to a structure that has a protective outer layer of lipid and can be used as a drug delivery vehicle.For example, lipid-based vehicle can be used to encapsulate and transport cargo (e.g., therapeutic agent) to biological target.Examples of lipid-based vehicle include, but are not limited to, liposome, micelle, polymerosome, and lipid nanoparticle.

[0182] As used herein, "biological molecule" refers to molecules such as proteins, nucleic acids, polysaccharides, and lipids.

[0183] The term "protein" is defined as a class of macromolecules containing long chains of one or more amino acids. A wide variety of proteins can be considered to belong to a protein family based on their similar structural features, specific biological functions, and / or association with specific microorganisms, particularly disease-causing microorganisms. Such proteins include, for example, antibodies, cytokines, chemokines, enzymes, hormones, vaccine antigens, cancer antigens, adjuvants, nutritional markers, and tissue-specific antigens.

[0184] As used herein, the term "nucleic acid" includes deoxyribonucleic acid (DNA), ribonucleic acid (RNA), messenger RNA (mRNA), small interfering RNA (siRNA), short hairpin RNA (shRNA), and microRNA (miRNA).

[0185] The term "antibody" refers to a large immunoglobulin protein produced by the immune system to recognize and neutralize foreign substances, such as pathogenic bacteria and viruses. The term "antibody" includes monoclonal antibodies (e.g., full-length or intact monoclonal antibodies), polyclonal antibodies, multivalent antibodies, multispecific antibodies (e.g., bispecific or trispecific antibodies, so long as they exhibit the desired biological activity), and can also include certain antibody fragments. Antibodies can be human, humanized, and / or affinity matured. An "antibody fragment" comprises only a portion of an intact antibody, which in certain embodiments retains at least one, and typically most or all, of the functions normally associated with that portion when present in an intact antibody. In one embodiment, an antibody fragment comprises the antigen-binding site of an intact antibody and thus retains the ability to bind antigen. In another embodiment, an antibody fragment, e.g., an antibody fragment comprising the Fc region, retains at least one of the biological functions normally associated with the Fc region when present in an intact antibody, such as FcRn binding, antibody half-life regulation, ADCC function, and complement fixation. In one embodiment, the antibody fragment is a monovalent antibody that has an in vivo half-life substantially similar to that of an intact antibody. For example, such an antibody fragment may comprise an antigen-binding arm linked to an Fe sequence, which can confer in vivo stability to the fragment.

[0186] The term "aggregation" refers to the formation of higher molecular weight amorphous species due to the non-covalent attachment ("clumping") of smaller species. The aggregation process can be irreversible or reversible. Many biological and synthetic molecules can undergo aggregation, including proteins, peptides, lipid particles, nucleic acids, inorganic nanoparticles, and organic nanoparticles (e.g., micelles, lipid nanoparticles, liposomes, polymerosomes) that may further contain encapsulated species.

[0187] In the case of protein aggregation, the formation of protein aggregates can be attributed to the inherent disordered nature of proteins or to misfolding of protein molecules, which results in the exposure of hydrophobic residues and surfaces that are normally buried within the interior of the protein's three-dimensional structure. Due to the hydrophobic effect, the exposed hydrophobic portions of misfolded proteins tend to interact with other misfolded protein molecules, shielding the exposed hydrophobic surfaces, which can lead to protein aggregation.

[0188] Some biological molecules are more "aggregation-prone" than others. For example, a protein's amino acid sequence and overall three-dimensional structure are related to its susceptibility to aggregation. For example, transmembrane proteins are more prone to aggregation (or aggregation-prone) than non-membrane proteins, especially when recombinantly expressed without stabilizers. Proteins exposed to conditions exceeding physiological conditions (37°C, near-neutral pH, isotonicity) may also be more prone to aggregation than in their native environment. Stress conditions such as temperature fluctuations, light, mechanical perturbations (e.g., rocking), surfaces, ultrasonic vibrations, pH changes, and changes in ionic strength can affect protein stability and induce aggregation. Protein aggregation can result in the formation of particles (i.e., precipitation) that may or may not be visible to the naked eye. The extent of protein aggregation, which is not visible to the naked eye, can be measured by various analytical methods known in the art, such as size exclusion chromatography (SEC), gel electrophoresis, asymmetric field-flow fractionation (AF4), analytical ultracentrifugation (AUC), mass spectrometry (MS), light microscopy, fluorescence microscopy, dynamic light scattering (DLS), multi-angle laser light scattering (MALLS), flow imaging, turbidity / nephelometry, and transmittance measurements.

[0189] As used herein, the term "reduced aggregation" of a biological molecule or lipid-based vehicle includes the reduction of all forms of aggregation. The degree or amount of aggregation observed (e.g., in a composition) can be reduced compared to the same biological molecule or lipid-based vehicle composition in the absence of the polyacrylamide-based copolymer of the present disclosure. Thus, "reduced aggregation" includes invisible aggregation or small amounts of aggregation (e.g., low levels of aggregated protein). Thus, the amount of aggregates present in a composition 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 biological molecule or lipid-based vehicle in the absence of the polyacrylamide-based copolymer. Aggregation can be measured by any method known in the art, including, but not limited to, size exclusion chromatography (SEC), gel electrophoresis, asymmetric field-flow fractionation (AF4), analytical ultracentrifugation (AUC), mass spectrometry (MS), light microscopy, fluorescence microscopy, dynamic light scattering (DLS), multi-angle laser light scattering (MALLS), flow imaging, turbidity / nephelometry, and transmittance measurements.

[0190] As used herein, the term "increased stability," when referring to a formulation containing a biological molecule or lipid-based vehicle, refers to a measurable decrease in the amount of aggregation over a period of time under test or fixed storage conditions compared to the amount of aggregate of the same biological molecule or lipid-based vehicle in the absence of the polyacrylamide-based copolymer.

[0191] As used herein, the term "aggregated protein" or "protein aggregate" refers to a collection of proteins that are disordered or misfolded and clump 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, films, filaments, pre-fibrils, amyloid deposits, amyloid fibrils, plaques, and dispersed non-natural intracellular oligomers. In some embodiments, proteins in protein aggregates are soluble precursors prior to their aggregation. Protein aggregation can be prevented in compositions containing polyacrylamide-based copolymers of the present disclosure. Protein aggregation can also be reduced in compositions containing polyacrylamide-based copolymers of the present disclosure compared to compositions containing the same protein without the polyacrylamide-based copolymers of the present disclosure. Thus, polyacrylamide-based copolymers can reduce or prevent protein aggregation.

[0192] 5. Additional Embodiments The present disclosure is further described by the following non-limiting clauses. 1. An injectable pharmaceutical composition comprising: A particle, Biopharmaceutical agents, and a particle comprising a polyacrylamide-based copolymer; and a liquid carrier in which the particles are suspended. 2. The injectable pharmaceutical composition of clause 1, wherein the particles have an average diameter of 100 microns or less. 3. The injectable pharmaceutical composition of clause 2, wherein the particles have an average diameter of 0.01 to 100 microns. 4. The injectable pharmaceutical composition of clause 3, wherein the particles have an average diameter of 0.1 to 100 microns. 5. The injectable pharmaceutical composition of clause 4, wherein the particles have an average diameter of 0.2 to 20 microns. 6. The injectable pharmaceutical composition of clause 5, wherein the particles have an average diameter of 0.2 to 10 microns. 7. The injectable pharmaceutical composition of clause 1, further comprising one or more of a stabilizer, a preservative, a filler, a bulking agent, a sugar, a polysaccharide, or a viscosity modifier. 8. The injectable pharmaceutical composition according to clause 7, further comprising a stabilizer. 9. The injectable pharmaceutical composition according to clause 8, wherein the stabilizer is selected from surfactants, poloxamers, povidone, polyvinylpyrrolidone (PVP) polymers, polyvinyl alcohol (PVA) polymers, polysaccharides, celluloses, amphoteric compounds, sugars, salts, and combinations thereof. 10. The injectable pharmaceutical composition of any one of clauses 1 to 9, wherein the biopharmaceutical agent is a polypeptide. 11. An injectable pharmaceutical composition according to clause 10, wherein the polypeptide is prone to aggregation in aqueous media. 12. The injectable pharmaceutical composition according to clause 10 or 11, wherein the polypeptide is selected from antibodies and fragments thereof, cytokines, chemokines, hormones, vaccine antigens, cancer antigens, adjuvants, and combinations thereof. 13. An injectable pharmaceutical composition according to any one of clauses 10 to 12, wherein the polypeptide is a protein. 14. The injectable pharmaceutical composition according to clause 13, wherein the protein is an antibody or a fragment thereof. 15. The injectable pharmaceutical composition of clause 14, wherein the protein is a monoclonal antibody, a polyclonal antibody, an immunoglobulin G (IgG) antibody, an IgA antibody, an IgM antibody, an Fc fusion protein, or a fragment thereof. 16. An injectable pharmaceutical composition according to any one of clauses 10 to 12, wherein the polypeptide is a hormone or an analog thereof. 17. The injectable pharmaceutical composition according to clause 16, wherein the polypeptide is insulin or an analog thereof. 18. The injectable pharmaceutical composition according to clause 16, wherein the polypeptide is selected from glucagon, a GLP-1 receptor agonist, amylin, and analogs thereof. 19. The injectable pharmaceutical composition according to any one of clauses 1 to 18, wherein the composition comprises 0.1 to 20% by weight of the biopharmaceutical agent. 20. The injectable pharmaceutical composition of clause 19, wherein the biopharmaceutical agent is insulin or an analog thereof. 21. The injectable pharmaceutical composition of clause 19, wherein the biopharmaceutical agent is a peptide. 22. The injectable pharmaceutical composition according to any one of clauses 19 to 21, wherein the particles comprise no more than 20% by weight of the biopharmaceutical agent. 23. The injectable pharmaceutical composition according to any one of clauses 1 to 18, wherein the composition comprises 20% by weight or more of the biopharmaceutical agent. 24. The injectable pharmaceutical composition of clause 23, wherein the composition comprises 20% to 80% by weight of the biopharmaceutical agent. 25. The injectable pharmaceutical composition of clause 23 or 24, wherein the biopharmaceutical agent is an antibody or fragment thereof or an Fc fusion protein. 26. An injectable pharmaceutical composition according to any one of clauses 23 to 25, wherein the particles comprise 75% by weight or more of the biopharmaceutical agent. 27. An injectable pharmaceutical composition according to any one of clauses 1 to 26, wherein the composition comprises 0.01 to 20% by weight of a polyacrylamide-based copolymer. 28. The injectable pharmaceutical composition according to clause 27, wherein the composition comprises 0.1 to 10% by weight of a polyacrylamide-based copolymer. 29. The injectable pharmaceutical composition according to clause 27, wherein the composition comprises 0.5 to 5% by weight of a polyacrylamide-based copolymer. 30. An injectable pharmaceutical composition according to any one of clauses 1 to 29, wherein the particles comprise 0.01 to 25% by weight of a polyacrylamide-based copolymer. 31. The injectable pharmaceutical composition according to clause 30, wherein the particles comprise 0.1 to 10% by weight of a polyacrylamide-based copolymer. 32. The injectable pharmaceutical composition according to clause 30, wherein the particles comprise 1 to 5% by weight of a polyacrylamide-based copolymer. 33. The injectable pharmaceutical composition of any one of clauses 1 to 32, wherein the composition comprises a weight to weight ratio of polyacrylamide-based copolymer to biopharmaceutical agent of 1:500 to 2:1. 34. The injectable pharmaceutical composition of clause 33, wherein the composition comprises a weight-to-weight ratio of polyacrylamide-based copolymer to biopharmaceutical agent of 1:50 to 1:1. 35. The injectable pharmaceutical composition of clause 33, wherein the composition comprises a weight-to-weight ratio of polyacrylamide-based copolymer to biopharmaceutical agent of 1:25 to 1:10. 36. A polyacrylamide copolymer is a water-soluble carrier monomer selected from N-(3-methoxypropyl)acrylamide (MPAM), 4-acryloylmorpholine (MORPH), N,N-dimethylacrylamide (DMA), N-hydroxyethylacrylamide (HEAM), acrylamide (AM), and combinations thereof; 36. The injectable pharmaceutical composition of any one of clauses 1 to 35, comprising 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-N-diethylacrylamide (DEA), N-tert-butylacrylamide (TBA), N-phenylacrylamide (PHE), and combinations thereof. 37. The injectable pharmaceutical composition according to clause 33, wherein the water-soluble carrier monomer is selected from MORPH, MPAM, and combinations thereof. 38. The injectable pharmaceutical composition according to clause 34, wherein the water-soluble carrier monomer comprises MORPH. 39. The injectable pharmaceutical composition according to clause 34, wherein the water-soluble carrier monomer comprises MPAM. 40. The injectable pharmaceutical composition according to any one of clauses 33 to 36, wherein the functional dopant monomer is selected from AMP, TMA, TBA, PHE, and combinations thereof. 41. The injectable pharmaceutical composition according to any one of clauses 33 to 36, wherein the functional dopant monomer is selected from DEA, PHE, NIP, and combinations thereof. 42. The injectable pharmaceutical composition according to any one of clauses 33 to 36, wherein the functional dopant monomer comprises a TRI. 43. The injectable pharmaceutical composition according to any one of clauses 33 to 36, wherein the functional dopant monomer comprises PHE. 44. The injectable pharmaceutical composition according to any one of clauses 33 to 36, wherein the functional dopant monomer comprises a NIP. 45. An injectable pharmaceutical composition according to any one of clauses 33 to 36, wherein the functional dopant monomer comprises DEA. 46. ​​The water-soluble carrier monomer is selected from MPAM, MORPH, and combinations thereof; 34. The injectable pharmaceutical composition of clause 33, wherein the functional dopant monomer is selected from NIP, PHE, and combinations thereof. 47. The water-soluble carrier monomer is selected from MPAM, MORPH, and combinations thereof; 34. The injectable pharmaceutical composition of clause 33, wherein the functional dopant monomer is selected from AMP, TMA, TBA, PHE, and combinations thereof. 48. The injectable pharmaceutical composition according to clause 33, wherein the water-soluble carrier monomer is MPAM and the functional dopant monomer is PHE. 49. The injectable pharmaceutical composition according to clause 33, wherein the water-soluble carrier monomer is MORPH and the functional dopant monomer is PHE. 50. The injectable pharmaceutical composition according to clause 33, wherein the water-soluble carrier monomer is MORPH and the functional dopant monomer is NIP. 51. A polyacrylamide copolymer is 70% to 98% by weight of a water-soluble carrier monomer; 51. The injectable pharmaceutical composition of any one of clauses 33 to 50, comprising 2% to 30% by weight of the functional dopant monomer. 52. A polyacrylamide copolymer is 80% to 95% by weight of a water-soluble carrier monomer; 51. The injectable pharmaceutical composition of any one of clauses 33 to 50, comprising 5% to 20% by weight of the functional dopant monomer. 53. A polyacrylamide copolymer is 83% to 98% by weight of a water-soluble carrier monomer; 51. The injectable pharmaceutical composition of any one of clauses 33 to 50, comprising 2% to 17% by weight of the functional dopant monomer. 54. A polyacrylamide copolymer is 70% to 85% by weight of MORPH; 34. The injectable pharmaceutical composition of clause 33, comprising 15% to 30% by weight of NIP. 55. A polyacrylamide copolymer is 74% to 80% by weight of MORPH; 20% to 26% by weight of NIP. 56. A polyacrylamide copolymer is 77% by weight of MORPH; 23% by weight of NIP. 57. An injectable pharmaceutical composition according to any one of clauses 33 to 56, wherein the degree of polymerization of the polyacrylamide copolymer is 10 to 500. 58. The injectable pharmaceutical composition according to clause 57, wherein the degree of polymerization of the polyacrylamide copolymer is 20-200. 59. An injectable pharmaceutical composition according to clause 58, wherein the degree of polymerization of the polyacrylamide copolymer is 50. 60. The injectable pharmaceutical composition according to any one of clauses 22 to 59, wherein the number average molecular weight of the polyacrylamide-based copolymer is between 1,000 g / mol and 40,000 g / mol. 61. The injectable pharmaceutical composition according to 60, wherein the number average molecular weight of the polyacrylamide-based copolymer is 2,000 g / mol to 10,000 g / mol. 62. The injectable pharmaceutical composition according to 61, wherein the number average molecular weight of the polyacrylamide copolymer is 4,000 g / mol to 6,000 g / mol. 63. An injectable pharmaceutical composition according to any one of clauses 1 to 62, wherein the polyacrylamide-based copolymer is amphiphilic. 64. An injectable pharmaceutical composition according to any one of clauses 1 to 63, wherein the liquid carrier comprises a mixture of an organic solvent and an aqueous solution. 65. The injectable pharmaceutical composition of any one of clauses 1 to 63, wherein the liquid carrier comprises an organic solvent, an oil, or a combination thereof. 66. The injectable pharmaceutical composition of any one of clauses 1 to 65, wherein the liquid carrier comprises one or more liquids selected from triacylglycerides, diacylglycerides, monoacylglycerides, acetamides, alkyl alcohols, aryl alcohols, aralkyl alcohols, fatty acids, fatty acid esters, oils, alkanes, perfluoroalkanes, propylene glycol monoesters, propylene glycol diesters, butylene glycol monoesters, butylene glycol diesters, polyethylene glycol diesters, and combinations thereof. 67. The injectable pharmaceutical composition of clause 66, wherein the liquid carrier comprises an aralkyl benzoate. 68. The injectable pharmaceutical composition according to clause 67, wherein the aralkyl benzoate is benzyl benzoate. 69. The injectable pharmaceutical composition of clause 66, wherein the liquid carrier comprises a triacylglyceride, a diacylglyceride, a monoacylglyceride, or a combination thereof. 70. The injectable pharmaceutical composition of clause 69, wherein the liquid carrier comprises a triacylglyceride which is triacetin. 71. The injectable pharmaceutical composition according to clause 66, wherein the liquid carrier comprises N,N-dimethylacetamide (DMAc). 72. The injectable pharmaceutical composition of clause 66, wherein the liquid carrier comprises a fatty acid ester. 73. The injectable pharmaceutical composition according to clause 72, wherein the fatty acid ester is ethyl oleate. 74. The injectable pharmaceutical composition according to clause 66, wherein the liquid carrier comprises a propylene glycol diester and / or a butylene glycol diester. 75. The injectable pharmaceutical composition according to clause 74, wherein the liquid carrier comprises Miglyol 840. 76. The injectable pharmaceutical composition of clause 65 or 66, wherein the liquid carrier comprises a blend of a triacylglyceride and one or more organic solvents having a lower viscosity than the triacylglyceride. 77. The injectable pharmaceutical composition according to clause 76, wherein the triacylglyceride is triacetin. 78. The injectable pharmaceutical composition according to clause 77, wherein the liquid carrier comprises a volume-to-volume (v / v) ratio of triacetin to one or more organic solvents having a lower viscosity than triacetin of 1:1 to 9:1. 79. The injectable pharmaceutical composition according to clause 76, wherein the liquid carrier comprises a 1:1 v / v ratio of triacetin to one or more organic solvents having a lower viscosity than triacetin. 80. The injectable pharmaceutical composition according to clause 76, wherein the liquid carrier comprises a 3:1 v / v ratio of triacetin to one or more organic solvents having a lower viscosity than triacetin. 81. The injectable pharmaceutical composition according to clause 76, wherein the liquid carrier comprises a 9:1 v / v ratio of triacetin to one or more organic solvents having a lower viscosity than triacetin. 82. The injectable pharmaceutical composition according to any one of clauses 76 to 81, wherein the organic solvent having a lower viscosity than triacetin is selected from acetamide, alkyl benzoate, aryl benzoate, aralkyl benzoate, aryl alcohol, aralkyl alcohol, or any combination thereof. 83. The injectable pharmaceutical composition according to clause 82, wherein the organic solvent having a lower viscosity than triacetin is acetamide. 84. The injectable pharmaceutical composition according to clause 83, wherein the acetamide is DMAc. 85. The injectable pharmaceutical composition according to clause 82, wherein the organic solvent having a lower viscosity than triacetin is an aralkyl alcohol. 86. The injectable pharmaceutical composition according to clause 85, wherein the aralkyl alcohol is benzyl alcohol. 87. The injectable pharmaceutical composition according to clause 82, wherein the organic solvent having a lower viscosity than triacetin is an aralkyl benzoate. 88. The injectable pharmaceutical composition according to clause 87, wherein the aralkyl benzoate is benzyl benzoate. 89. The injectable pharmaceutical composition according to clause 82, wherein the organic solvent having a lower viscosity than triacetin comprises a combination of acetamide and an aralkyl alcohol. 90. The injectable pharmaceutical composition according to clause 89, wherein the acetamide is DMAc and the aralkyl alcohol is benzyl alcohol. 91. The injectable pharmaceutical composition according to clause 90, wherein the low viscosity organic solvent comprises a v / v ratio of DMAc to benzyl alcohol of 3:1 to 2:1. 92. The injectable pharmaceutical composition according to clause 65 or 66, wherein the organic solvent is an alkyl benzoate, an aryl benzoate, an aralkyl benzoate, or any combination thereof. 93. The injectable pharmaceutical composition according to clause 92, wherein the organic solvent is benzyl benzoate. 94. An injectable pharmaceutical composition according to clause 64 or 65, wherein the organic solvent is a polar aprotic solvent. 95. The injectable pharmaceutical composition according to clause 94, wherein the polar aprotic solvent is dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), or a mixture thereof. 96. The injectable pharmaceutical composition according to clause 65, wherein the liquid carrier comprises an oil (e.g., a vegetable oil or an animal oil). 97. The injectable pharmaceutical composition according to clause 96, wherein the oil is coconut oil, cottonseed oil, fish oil, grapeseed oil, hazelnut oil, hydrogenated vegetable oil, lime oil, olive oil, palm seed oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, walnut oil, or any combination thereof. 98. The injectable pharmaceutical composition according to clause 97, wherein the oil is sesame oil. 99. An injectable pharmaceutical composition according to any one of clauses 1 to 98, wherein the composition comprises a weight to weight ratio of liquid carrier to particles of 1:5 to 4:1. 100. The injectable pharmaceutical composition according to clause 99, wherein the composition comprises a weight to weight ratio of liquid carrier to particles of 1:1 to 2:1. 101. An injectable pharmaceutical composition comprising: A particle, the particle comprising: Biopharmaceutical agents, and a polyacrylamide-based copolymer, wherein 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), acrylamide (AM), and combinations thereof; and particles comprising 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-N-diethylacrylamide (DEA), N-tert-butylacrylamide (TBA), N-phenylacrylamide (PHE), and combinations thereof; an injectable pharmaceutical composition comprising: a liquid carrier in which the particles are suspended, the liquid carrier being selected from a triacylglyceride, a diacylglyceride, a monoacylglyceride, an acetamide, an alkyl alcohol, an aryl alcohol, an aralkyl alcohol, a fatty acid, a fatty acid ester, an oil, an alkane, a perfluoroalkane, a propylene glycol monoester, a propylene glycol diester, a butylene glycol monoester, a butylene glycol diester, a polyethylene glycol diester, and combinations thereof. 102. The injectable pharmaceutical composition according to clause 101, wherein the water-soluble carrier monomer is selected from MORPH, MPAM, and combinations thereof. 103. The injectable pharmaceutical composition according to any one of clauses 101-102, wherein the functional dopant monomer is selected from AMP, TMA, TBA, PHE, and combinations thereof. 104. The injectable pharmaceutical composition according to any one of clauses 101-102, wherein the functional dopant monomer is selected from DEA, PHE, NIP, and combinations thereof. 105. The water-soluble carrier monomer is selected from MPAM, MORPH, and combinations thereof; 105. The injectable pharmaceutical composition of clause 104, wherein the functional dopant monomer is selected from NIP, PHE, and combinations thereof. 106. The injectable pharmaceutical composition according to clause 104, wherein the water-soluble carrier monomer is MORPH and the functional dopant monomer is NIP. 107. An injectable pharmaceutical composition according to any one of clauses 101 to 106, wherein the liquid carrier comprises an aralkyl benzoate. 108. The injectable pharmaceutical composition according to clause 107, wherein the aralkyl benzoate is benzyl benzoate. 109. An injectable pharmaceutical composition according to any one of clauses 101 to 106, wherein the liquid carrier comprises a triacylglyceride. 110. The injectable pharmaceutical composition according to clause 109, wherein the triacylglyceride is triacetin. 111. An injectable pharmaceutical composition according to any one of clauses 101 to 106, wherein the liquid carrier comprises N,N-dimethylacetamide (DMAc). 112. An injectable pharmaceutical composition according to any one of clauses 101 to 106, wherein the liquid carrier comprises a fatty acid ester. 113. The injectable pharmaceutical composition according to clause 112, wherein the fatty acid ester comprises ethyl oleate. 114. An injectable pharmaceutical composition according to any one of clauses 101 to 106, wherein the liquid carrier comprises a propylene glycol diester and / or a butylene glycol diester. 115. The propylene glycol diester is Miglyol 840; 115. The injectable pharmaceutical composition according to clause 114, wherein the butylene glycol diester is Miglyol 8810. 116. A syringe loaded with a composition according to any one of clauses 1 to 115. 117. The syringe of clause 116, configured to dispense the composition at a flow rate of 0.1 mL / min or greater in response to a force applied to the syringe of 70 N or less. 118. The syringe of clause 116, configured to dispense the composition at a flow rate of 0.1 mL / min or greater in response to a force applied to the syringe of 50 N or less. 119. A syringe according to any one of clauses 116 to 118, wherein the syringe is equipped with a needle having a size of 18 to 32 gauge (G). 120. A syringe according to clause 119, wherein the needle is a 22G needle. 121. A syringe according to clause 119, wherein the needle is a 24G needle. 122. A syringe according to clause 119, wherein the needle is a 25G needle. 123. A syringe according to clause 119, wherein the needle is a 26G needle. 124. A syringe according to clause 119, wherein the needle is a 27G needle. 125. A syringe according to clause 119, wherein the needle is a 30G needle. 126. A syringe according to clause 119, wherein the needle is a 32G needle. 127. A syringe according to any one of clauses 119 to 126, wherein the needle is an ultra-thin-walled needle. 128. A method for administering a biopharmaceutical to a subject by injection, comprising: 116. A method comprising injecting a composition according to any one of clauses 1 to 115 to administer a therapeutically effective dose of a biopharmaceutical to a subject in need thereof. 129. The method of clause 128, wherein the injecting is performed using a loaded syringe according to any one of clauses 116 to 127. 130. A method for preparing an injectable pharmaceutical composition, comprising: a) a biopharmaceutical agent; providing a mixture comprising: a polyacrylamide-based copolymer in water; b) removing water from the mixture to obtain particles comprising the biopharmaceutical agent and the polyacrylamide-based copolymer; c) contacting the liquid carrier with the particles to form a suspension of the particles in the liquid carrier. 131. The method of clause 130, wherein in step b), water is removed via spray drying. 132. The method of clause 130, wherein in step b), water is removed via electrospray drying. 133. The method according to clause 130, wherein in step b) water is removed via freeze-drying. 134. The method according to clause 130, wherein the method further comprises a milling step after freeze-drying but before step c). 135. The method of any one of clauses 130 to 134, wherein the particles obtained in step b) have an average diameter of 100 microns or less. 136. The method of any one of clauses 130-135, wherein step c) comprises sonicating the particles in the liquid carrier. [Example]

[0193] 6. Working Example The examples in this section are provided by way of illustration, not limitation. The examples may represent only some embodiments, and it should be understood that the following examples are illustrative and not limiting. Unless otherwise specified, all substituents are as previously defined. Reagents and starting materials are readily available to those skilled in the art. The specific synthetic steps for each of the routes described can be combined in different ways or with steps from different schemes to prepare the compounds and compositions described herein.

[0194] General method All reagent-grade materials and solvents were purchased from Sigma-Aldrich or Fisher and used as received. Alexa-647-NHS was purchased from Lumiprobe. Slide-A-Lyzer dialysis cassettes (2 kDa MWCO) from Thermo Fisher were used for polymer purification. BSA (A2153-50G, CAS-No. 9048-46-8) was purchased as a lyophilized powder from Sigma-Aldrich. Human IgG (Cat. No. 340-21, Lot: 07J4627) was purchased as a lyophilized powder from Medix Biochemica. HyPure™ cell culture-grade water was purchased from Cytiva. Phosphate-buffered saline (10010-023) was purchased from Gibco. The syringe used for injection force measurements was a Fisherbrand 1 mL plastic Luer-Lok syringe (Cat. No. 14955464). The syringe used to formulate the protein suspension with the non-solvent was a Thermo Scientific 5 mL leur slip plastic syringe (catalog number: S7510-5). The needle used for injection force measurements was a BD PrecisionGlide™ needle (26 G, ½ inch, reference: 305111). In vivo protein suspension delivery was performed using a BD insulin syringe with a BD Micro-Fine™ IV needle (28 G, 12.7 mm).

[0195] statistical analysis Injection force data are reported as means with standard deviations. For in vivo experiments, animals were cage-blocked, and Mead's resource equation was used to determine the above sample size, where additional subjects have little effect on power. Normalized fluorescence intensity and subcutaneous absorption half-life from in vivo experiments are reported as means with standard errors. Comparisons between groups were performed using Tukey's HSD test in JMP. Results were considered significant when p<0.05.

[0196] 6.1.1. Example 1 - Synthesis of Polyacrylamide-Based Copolymer General method Polyacrylamide-Based Copolymer Synthesis: Polyacrylamide-based copolymers can be synthesized using any convenient method. Methods that can be used or adapted for use in preparing the copolymers of the present disclosure include the exemplary synthetic method described herein in Example 1.1, as well as those described by Appel et al. in PCT Application PCT / US2021 / 027693, filed April 16, 2021, and Mann et al., Sci. Transl. Med. 12, eaba6676 (2020), the disclosures of which are incorporated herein by reference in their entirety.

[0197] Copolymer molecular weight characterization: Mn, Mw, and dispersity for copolymers with HEAM, DMA, MPAM, and MORPH carrier monomers are determined via SEC implementing poly(ethylene glycol) standards (American Polymer Standards Corporation) after passing through two size exclusion chromatography columns.

[0198] The Mn, Mw, and dispersity of the copolymer with AM are determined via SEC-MALLS after passing through a size-exclusion chromatography column in a mobile phase of phosphate-buffered saline containing 300 ppm sodium azide. Detection is performed with an Optilab T-rEX (Wyatt Technology Corporation) refractive index detector operating at 658 nm and a TREOS II light scattering detector (Wyatt Technology Corporation) operating at 659 nm. The dn / dc value for the AM copolymer is assumed to be 0.185 in this medium.

[0199] Example 1.1 - Synthesis of MoNi Amphiphilic acrylamide copolymer excipient 4-acryloylmorpholine 77% -N-Isopropylacrylamide 23%(MoNi77:23, also referred to herein as "MoNi") was prepared according to the method described by Mann et al., Sci. Transl. Med. 12, eaba6676 (2020). MORPH (645 mg, 4.57 mmol, 41.5 equiv.), NIP (105 mg, 0.93 mmol, 8.5 equiv.), RAFT CTA 2-cyano-2-propyldodecyltrithiocarbonate (2-CPDT) (38 mg, 0.11 mmol, 1 equiv.), and initiator 2,2-azobis(2-methylpropionitrile)AIBN (3.6 mg, 0.02 mmol, 0.2 equiv.) were combined and diluted with N,N-dimethylformamide (DMF) to a total volume of 2.25 mL (vinyl monomer concentration of 33.3 w / v) in an 8 mL scintillation vial equipped with a PTFE septum. The reaction mixture was sparged with nitrogen gas for 10 min and then heated at 65 °C for 12 h. To remove the CTA Z-terminus of the resulting polymer, AIBN (360 mg, 2.2 mmol, 20 eq.) and lauroyl peroxide (LPO) (88 mg, 0.22 mmol, 2 eq.) were added to the reaction mixture, which was then sparged with nitrogen gas for 10 minutes and heated at 90°C for 12 hours. CTA Z-group removal was confirmed by the ratio of refractive index to ultraviolet (310 nm) intensity in size exclusion chromatography (SEC) analysis. The resulting polymer was precipitated from ether three times and dried under vacuum overnight. The resulting composition and molecular weight were determined by: 1 Determined via 1 H NMR spectroscopy (e.g., in d6-DMSO) and SEC using PEG standards.

[0200] Example 1.2 - Characterization of Copolymer Molecular Weight by SEC M about MoNi n , M w , and D are at 35°C and 1.0 mL min -1 Two SEC columns [7.8 mm i.d., Mw range 200–600,000 g mol] were run in a mobile phase of DMF with 0.1 M LiBr at a flow rate of 100 s.p.m. -1, Resolve Mixed Bed Low divinylbenzene (DVB) (Jordi Labs)] and then determined via SEC implementing PEG standards (American Polymer Standards Corporation) [Dionex UltiMate3000 pump, degasser, and autosampler (Thermo Fisher Scientific)].

[0201] The MoNi polymer was also characterized for molecular weight (M n SEC The chromatographic analysis was characterized by determining the chromatographic density and dispersity. The running solvent was N,N-dimethylformamide (DMF) with 1 g / L LiBr (flow rate, 1 mL / min) heated to 50 °C, and samples were prepared at 5 mg / mL. Separation was performed sequentially through two Jordi Labs Resolve Mixed Bed Low Divinylbenzene (DVB) columns, and data were collected using a Dionex Ultimate 3000 Variable Wavelength detector and a RefractoMax521 RI detector. RI traces were normalized, and the area under the curve of the 310 nm absorbance signal was calculated using Prism 10.

[0202] Figure 1, panels a-c, illustrate the characterization of MoNi. Panel a depicts the SEC trace of MoNi. Panel b depicts the SEC trace of MoNi. 1 Panel c depicts the H NMR. Panel c depicts the DSC of MoNi at a temperature ramp and cooling rate of 10 °C / min, showing a glass transition temperature of 130–140 °C (top line = cooling, bottom line = heating).

[0203] 6.1.2. Example 2 - Preparation of Particles and Particle Suspensions Particles and particle suspensions comprising the subject polyacrylamide-based copolymers and protein agents were prepared by one of the following methods.

[0204] Example 2.1 - General Method A - Freeze-Drying and Ball Milling The protein agent in water is optionally combined with the subject polyacrylamide copolymer and buffer solution.Then, water is removed by freeze-drying, and the resulting mixture is milled to obtain particles.Then, a liquid carrier (for example, as described herein) is added to the particles by mixing, to obtain a suspension of particles in the liquid carrier.

[0205] Example 2.2 - General Method B - Spray Drying The protein agent in water or aqueous organic solvent mixture is optionally combined with the subject polyacrylamide copolymer.Then, the water or aqueous organic solvent mixture is removed by spray drying to provide particles.Then, a liquid carrier (for example, as described herein) is added to the particles by mixing, to provide a suspension of particles in the liquid carrier.

[0206] FIG. 2 illustrates a schematic of the spray drying process and the formulation of an exemplary particle suspension.

[0207] Method B1 - Spray-drying BSA particles: A BSA feed solution was prepared by dissolving lyophilized BSA in 2% by weight (20 mg / mL) cell-grade water. The BSA solution was allowed to dissolve at room temperature for 1 hour and then sterile-filtered using a 0.2 μm sterile filter. After sterile filtration, 7 kDa MoNi was added to the feed solution at a concentration of 0.1% by weight (1 mg / mL). The feed solution was stored on ice and then spray-dried.

[0208] The samples were spray dried using a Buchi B-290 Mini Spray Dryer equipped with a high-efficiency cyclone. Samples were spray dried using an inlet temperature of 150°C (outlet approximately 67°C), an aspirator pressure of 40 mm, and a pump speed of 20% (6 mL / min). The collected particles were transferred to a 50 mL Falcon tube and stored at 4°C with desiccant.

[0209] Method B2 - Spray-drying BSA particles with polysorbate 80 (Tween 80): A BSA feed solution was prepared by dissolving lyophilized BSA in 2% by weight (20 mg / mL) cell-grade water. The BSA solution was allowed to dissolve at room temperature for 1 hour before sterile filtering using a 0.2 μm sterile filter. After sterile filtration, polysorbate 80 was added to the feed solution at a concentration of 0.0176% by weight (0.176 mg / mL). The polysorbate 80 concentration was selected so that equimolar amounts of MoNi and polysorbate 80 were added to the spray-dried feed solution. The feed solution was stored on ice before being spray-dried.

[0210] The samples were spray dried using a Buchi B-290 Mini Spray Dryer equipped with a high-efficiency cyclone. Samples were spray dried using an inlet temperature of 150°C (outlet approximately 67°C), an aspirator pressure of 40 mm, and a pump speed of 20% (6 mL / min). The collected particles were transferred to a 50 mL Falcon tube and stored at 4°C with desiccant.

[0211] Method B3 - Spray drying of human immunoglobulin G (hIgG) particles: The hIgG feed solution was prepared by dissolving lyophilized hIgG in cell-grade water at 10% by weight (100 mg / mL). The hIgG solution was dissolved at 4°C for 4 hours and then sterile filtered using a 0.2 μm sterile filter. Protein deposition on the sterile filter was minimized by filtering 2% by weight of BSA through the sterile filter, followed by five water rinses. Nanodrop™ confirmed that no BSA was detectable in the filtrate. After sterile filtering the hIgG, 7 kDa MoNi was added to the feed solution at a concentration of 0.5% by weight (5 mg / mL). The feed solution was stored on ice and then spray-dried.

[0212] The samples were spray dried using a Buchi B-290 Mini Spray Dryer equipped with a high-efficiency cyclone. Samples were spray dried using an inlet temperature of 130°C (outlet approximately 77°C), an aspirator pressure of 40 mm, and a pump speed of 5% (2 mL / min). The collected particles were transferred to a 50 mL Falcon tube and stored at 4°C with desiccant.

[0213] Particle characterization: Particle morphology was characterized by scanning electron microscopy (SEM). Samples were grounded to aluminum pin stubs using double-sided conductive copper tape. A 5.0 nm thick layer of pure gold was deposited on the samples using a Leica ACE600 Vacuum system. SEM analysis was performed using an FEI Magellan 400XHR scanning electron microscope at high vacuum, 5.00 kV, and in field-free mode.

[0214] Particle density was measured using an AccuPyc 1330. A known sample mass of 200-300 mg of spray-dried particles was measured using a 1 cm 3 The particle volume was measured over 999 cycles. The particle density was calculated using the known mass measured by the analytical balance and the average sample volume.

[0215] Formulating suspensions: Suspensions were formulated by combining a known mass of spray-dried particles with a known volume of non-solvent. Protein concentrations in mg / mL were determined by assuming the total volume included the non-solvent volume and the spray-dried particle volume. Unless otherwise specified, protein particles were concentrated at 1 g / cm. 3 It was assumed that the density of

[0216] To minimize nonsolvent evaporation during suspension preparation, the spray-dried particles were added to the barrel of a 6 mL Luer-slip syringe. The mass of the spray-dried particles was measured using an analytical balance. The desired volume of nonsolvent or nonsolvent combination was added to the syringe barrel through the syringe tip using a p200 pipette. After capping the syringe, the protein suspension was mixed inside the syringe using a vortex for 5 minutes or until all powder was completely dispersed. The protein suspension was transferred from the 6 mL Luer-slip syringe to the desired alternative syringe (1 mL Luer-lock syringe or insulin syringe) by backloading for injection force experiments or animal experiments, respectively.

[0217] Example 2.3 - Characterizing the flow properties of particle suspensions The flow properties of the particle suspension were characterized through rheology and injection force measurements.

[0218] Rheological characterization: Rheological testing was performed using a stress-controlled TA Instruments DHR-2 rheometer. Rheology of solid-like formulations (BSA without MoNi) was performed at 25°C using 20 mm diameter sawtooth parallel plates with a 500 μm gap. Rheology of liquid-like formulations (BSA with MoNi) was performed at 25°C using a 40 mm cone geometry with a 50 μm gap. Frequency sweeps were performed at 1% strain within the linear viscoelastic region. Flow sweeps were performed from high to low shear rates using steady-state detection.

[0219] Injection force measurement: Injection force was quantified by measuring the force required to inject a protein particle suspension through a known needle gauge at a known flow rate using a syringe with known barrel dimensions. A force sensor was constructed incorporating a load cell (FUTEK LLB300 50 lb Subminiature Load Button (Model Number: LLB300, Item Number: FSH03954, Serial Number: 705242)) attached to a syringe pump (KD Scientific Syringe Pump (Model Number: LEGATO100, Catalog Number: 788100, Serial Number: D103954)). Load cell resistance measurements were converted to force values ​​in kg using an Omega Engineering Platinum Series Meter (Model Number: DP8PT, Serial Number: 18110196). Prior to measuring injection force, the load cell was calibrated. The LabView program recorded the measured force throughout the duration of the injection experiment and displayed a graph of injection force over time.

[0220] Injection force experiments were performed as follows: A 1 mL Thermo Fisher Luer-Lock syringe with the desired needle gauge was loaded into a syringe pump. The syringe pump height was adjusted so that the load button on the force sensor was in contact with the end of the syringe plunger. The initial force was at or very close to 0 kg. The appropriate syringe barrel dimensions, as well as the desired flow rate and injection volume, were then selected. The syringe pump moved at the programmed speed and injected the protein suspension through the attached needle. A force sensor coupled to the Omega unit measured the force required to inject the protein suspension at the desired flow rate. The LabView program recorded the measured force throughout the duration of the injection experiment and displayed a graph of the injection force over time. The injection force was quantified by subtracting the average initial force (background) from the average plateau injection force. The injection force in kg was converted to injection force in Newtons by multiplying by 9.81.

[0221] Exemplary Formulations: Tables 1 and 2 below provide formulations obtained by General Method A and General Method B, respectively.

[0222] Table 1: Bovine serum albumin (BSA) injectable formulations prepared by general method A: [Table 1]

[0223] Formulations 2, 4, 5, and 7 contain the polyacrylamide-based copolymer MoNi. Formulations 1, 3, and 6, and comparable injectable formulations, do not contain the subject polyacrylamide-based copolymer.

[0224] Referring to Formulations 6 and 7, as seen in Table 1, Formulation 7, which contains the subject copolymer MoNi as the polymer additive in the particles and a total solids content of 460 mg / mL, is injectable through both 21G and 26G needles, whereas Formulation 6, which has a lower total solids content and no polymer additive, is injectable through a 21G needle but not a 26G needle.

[0225] Table 2: Bovine serum albumin (BSA) injectable formulations prepared by general method B: [Table 2]

[0226] Formulations 9 and 11 contain the polyacrylamide-based copolymer MoNi. Formulations 8 and 10, and comparable injectable formulations, do not contain the subject polyacrylamide-based copolymer.

[0227] With reference to formulations 8 and 9, as seen in Table 2, formulation 9, which contains the subject copolymer MoNi as a polymer additive and a total solids content of 442 mg / mL, is injectable through both 27G and 30G needles, whereas formulation 8, which has a lower total solids content and does not contain any polymer additive, is injectable through a 27G needle but not a 30G needle. Similarly, with reference to formulations 10 and 11, as seen in Table 2, formulation 11, which contains the subject copolymer MoNi as a polymer additive and a total solids content of 486 mg / mL, is injectable through a 27G needle, whereas formulation 10, which has a lower total solids content and does not contain any polymer additive, is not injectable through a 27G needle.

[0228] These results demonstrate that the addition of small amounts of the subject copolymers (e.g., MoNi) to protein formulations (e.g., BSA) provides improved injectable properties despite higher solids contents.

[0229] Figure 3, panels a-c, illustrate the injection behavior of various formulations. Panel a illustrates injection through a 21G needle and the depot formation behavior of Formulation 4 (336 mg / mL BSA in sesame oil with copolymer MoNi and trehalose, 497 mg / mL total solids). Panel b illustrates injection through a 21G needle and the lack of depot formation behavior of Formulation 3 (364 mg / mL BSA in sesame oil with trehalose, but no copolymer, 455 mg / mL total solids). Panel c illustrates injection through a 26G needle and the depot formation behavior of Formulation 7 (400 mg / mL BSA in triacetin with copolymer MoNi and trehalose, 460 mg / mL total solids).

[0230] As illustrated in Figure 1, panels a-c, the addition of the subject copolymer MoNi modifies the injection and rheological properties of the formulation.

[0231] Figure 4, panels a-d, illustrate angular frequency sweeps and flow sweeps of formulations 6 and 7. Panel a illustrates the angular frequency sweep of formulation 7 (400 mg / mL BSA in triacetin with MoNi). Panel b illustrates the flow sweep of formulation 7 (400 mg / mL BSA in triacetin with MoNi). Panel c illustrates the angular frequency sweep of formulation 6 (400 mg / mL BSA in triacetin without copolymer). Panel d illustrates the flow sweep of formulation 6 (400 mg / mL BSA in triacetin without copolymer).

[0232] Figure 5, panels a-c, illustrate the rheological and stability characterization of ball-milled BSA particles. Panel a illustrates the angular frequency sweep of i) 400 mg / mL BSA particles resuspended in triacetin and ii) 400 mg / mL BSA particles containing 5 wt% MoNi resuspended in triacetin. Panel a.iii) Comparative storage modulus of the suspensions at 10 rad / s. Panel b illustrates microscopy images of BSA with 5 wt% MoNi particles formed by i) 15 min of ball-milling or ii) spray-drying. The particles are resuspended in sesame oil for improved imaging. Panel c depicts SEC traces of a fresh BSA control, 15 min of ball-milled BSA without MoNi, and 15 min of ball-milled BSA with 5 wt% MoNi. PBS with sodium azide was used as the eluent.

[0233] Figure 6, panels a-d, illustrate angular frequency sweeps and flow sweeps of formulations 10 and 11. Panel a illustrates the angular frequency sweep of formulation 11 (460 mg / mL BSA in triacetin with MoNi). Panel b illustrates the flow sweep of formulation 11 (460 mg / mL BSA in triacetin with MoNi). Panel c illustrates the angular frequency sweep of formulation 10 (460 mg / mL BSA in triacetin with no copolymer). Panel d illustrates the flow sweep of formulation 10 (460 mg / mL BSA in triacetin with no copolymer).

[0234] Figure 7, panels a-d, illustrate the rheological and stability characterization of spray-dried particles. Panel a illustrates the angular frequency sweep of i) 460 mg / mL BSA particles resuspended in triacetin and ii) 460 mg / mL BSA particles containing 5 wt% MoNi resuspended in triacetin. Panel a.iii) Comparative storage modulus of the suspensions at 10 rad / s. Panel b illustrates the flow sweep of 460 mg / mL BSA particles resuspended in triacetin and 460 mg / mL BSA particles containing 5 wt% MoNi resuspended in triacetin. Panel c shows SEM images (scale bar 5 μm) of the particles and the resulting suspension in 460 mg / mL triacetin for particles formulated i) without and ii) with 5 wt% MoNi. Panel d depicts the SEC traces of fresh BSA control, spray-dried BSA without MoNi, and spray-dried BSA with 5 wt% MoNi. PBS with sodium azide is used as the eluent.

[0235] Rheological characterization of protein particle suspensions in triacetin demonstrates that the addition of 5 wt% MoNi to the protein particles improves the flow properties of the protein suspension. As shown in Figure 7, protein suspensions were prepared at 460 mg / mL BSA using particles with and without 5 wt% MoNi. Without the addition of MoNi, the protein suspension had the consistency of a thick, solid paste and did not flow; however, with the addition of MoNi, the protein suspension instead flowed like a liquid. Angular frequency sweeps demonstrate that the addition of MoNi reduced the storage and loss moduli of the suspension by two orders of magnitude. Additionally, the tan delta of the suspension with MoNi exceeded 1, indicating liquid-like behavior, while the suspension without MoNi had a tan delta below 1, indicating solid-like behavior (Figure 7, panel a). Flow sweeps further demonstrate that the protein suspension shear thins and that MoNi results in a reduction in suspension viscosity (Figure 7, panel b). SEM images of spray-dried microparticles illustrate that while microparticles spray-dried with and without MoNi have similar average diameters (5-10 μm), the particles with MoNi have a smooth spherical morphology, while the particles without MoNi exhibit a wrinkled surface morphology (Figure 7, panel c). This difference in spray-dried particle morphology in the presence of MoNi surfactant may contribute to a reduction in the yield stress of the protein suspension, improving its flow properties. In addition to flow, MoNi also stabilizes BSA throughout the high-temperature spray-drying process. SEC characterization of fresh BSA and spray-dried BSA with and without MoNi shows that spray-drying BSA with MoNi results in a higher monomer peak fraction and a smaller high-molecular-weight aggregate peak (Figure 7, panel d). These improvements in BSA stability and suspension flow with MoNi were also observed when protein microparticles were instead formed by freeze-drying followed by ball milling (Figure 5, panels a-c).

[0236] 6.1.3. Example 3 - Characterization of protein stability in injectable formulations formed by Method A. To investigate protein stability in the subject injectable formulations, the formulations in Table 3 were prepared by general method A. [Table 3]

[0237] Figure 8, panels a-b, illustrates the effect of trehalose and copolymer MoNi on BSA stability after lyophilization (panel a) and milling (panel b). Panel a demonstrates that trehalose and MoNi content during lyophilization have little effect on the high molecular weight shoulder. Panel b further demonstrates that MoNi, regardless of trehalose content, reduces the size of the high molecular weight shoulder after 15 minutes of ball milling.

[0238] 6.1.4. Example 4 - Characterization of protein stability after spray drying and comparison with ball milling. Protein stability before and after spray drying was characterized by SEC. Protein suspension stability was characterized by SEC and comparative injection force.

[0239] Example 4.1 - Characterization of Protein Stability SEC characterization: SEC traces of protein samples were determined with the ASTRA software package (Wyatt Technology Corporation) after passing 5 mg / mL protein samples through a size-exclusion chromatography column (Superose6 Increase 10 / 300GL) in a mobile phase of PBS with sodium azide at 25 °C and a flow rate of 0.5 mL / min. Detection consisted of 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. A dn / dc value of 0.185 was used for BSA and IgG samples. To directly compare the stability of spray-dried protein samples, SEC traces were normalized to the height of the monomer peak.

[0240] Stress-aging methodology: BSA protein suspensions were prepared at 460 mg / mL in triacetin using spray-dried BSA particles with 5 wt% MoNi and BSA particles without 5 wt% MoNi. A BSA aqueous control was prepared by dissolving fresh lyophilized BSA in PBS at 20 mg / mL. All samples were stored in parafilmed 8 mL scintillation vials. A 500 mL beaker of water was heated to 60 °C using a temperature-controlled hot plate. The sample file was immersed in a 60 °C water bath so that the protein sample volume was completely below the water line. The sample was heated at 60 °C for 30 minutes to promote proteolysis. After stress-aging, the sample was redissolved in PBS at 5 mg / mL, and protein stability was evaluated via SEC.

[0241] Protein Suspension Storage and Injection Force Measurement: A BSA protein suspension with 5 wt% MoNi was prepared at 460 mg / mL in triacetin. The protein suspension was loaded into a 1 mL Thermo Fisher Luer slip syringe and capped with a BD26G ½ inch needle wrapped in parafilm to limit solvent evaporation. The injection force of the 460 mg / mL protein suspension was measured on day 0 and again on day 35. The syringe was stored horizontally at 23°C.

[0242] Example 4.2 - Comparison of spray dried and ball milled particles Exemplary BSA protein particle compositions prepared via spray drying or ball milling were evaluated for protein stability using size exclusion chromatography (SEC).

[0243] Figure 9, panels a(i)-a(iii) and b, show the results of evaluating exemplary particle compositions for protein stability using SEC. The improved resolution of the SEC column allows visualization of the BSA monomer and dimer peaks. Panel a(i): Full SEC traces of fresh, spray-dried, and ball-milled BSA. Panel a(ii): Dimer peaks of fresh, spray-dried, and ball-milled BSA. Panel a(iii): Fresh, spray-dried particles with a 100:5 weight ratio of BSA, and the high molecular weight peak of ball-milled BSA. Panel b: BSA monomer fraction after spray drying and ball-milling.

[0244] Fresh BSA and spray-dried BSA with and without MoNi show nearly identical SEC traces, indicating that BSA is stable throughout the spray-drying process. Spray-dried BSA with MoNi has a slightly higher monomer fraction, suggesting improved stability. Ball-milled BSA exhibits a higher dimer peak than spray-dried BSA, and the dimer peak of ball-milled BSA with MoNi is smaller than that of ball-milled BSA alone. These findings suggest that spray-drying is a gentler process that forms particles than ball-milling, but that MoNi is a useful protectant throughout the ball-milling process. SEC traces were obtained using a Superose 6 column. PBS with sodium azide was used as the eluent.

[0245] 6.1.5. Example 5 - Injectability of suspension formulations prepared by general method B To investigate the injectability of the subject formulations, injection force measurements were performed on Formulation 11 (containing spray-dried particles with a 100:5 ratio of BSA to copolymer MoNi in triacetin at a total protein concentration of 460 mg / mL).

[0246] Figure 10, panels a-c and d(i)-d(ii). Panel a: A force sensor connected to a syringe pump was used to quantify the injection force through a standard gauge needle using a 1 mL syringe. Panel b: The injection force curves illustrate the injection force required to inject a composition containing 460 mg / mL BSA and 23 mg / mL MoNi composition (i.e., 5% by weight of particle solids) through a 27G ½ inch needle at various flow rates. The BSA concentration assumes a particle density of 1 g / cm^3. For BSA particle density, see Example 11 below. The BSA concentration is estimated to be close to 519.5 mg / mL. Panel c: The injection force is linear with flow rate and varies with needle gauge. Panel d(i): Injection force curves illustrate the injection force required to inject a composition containing 460 mg / mL BSA and 23 mg / mL MoNi (i.e., 5% by weight of suspended particle solids) through a 27G ½ inch needle and a 26G ½ inch needle at 1 mL / min. Panel d(ii): Injection force decreases as needle gauge increases. An injection force comparison with a BSA formulation without MoNi is not included because the formulation without MoNi is not injectable through a 26 or 27G needle.

[0247] Figure 11, panels a-c. (Panel a) Further illustrating that i) injection force is linear with flow rate and ii) varies with needle gauge. (Panel b) Illustration of 460 mg / mL BSA with 5% MoNi in triacetin after 120 hours of vertical storage in a syringe. (Panel c) Injection force of 460 mg / mL BSA with 5% MoNi in triacetin on days 0 and 35 using a 26G ½ inch needle and a 1 mL / min flow rate.

[0248] For therapeutic applications, protein suspensions should be injectable through clinically relevant needle gauges with clinically relevant injection forces. To evaluate the injectability of a suspension of BSA with MoNi in triacetin, the injection force required to inject a 460 mg / mL BSA suspension through a 27-gauge, 1 / 2-inch needle at a given flow rate was quantified using a force sensor attached to a syringe pump (Figure 10, panel a). The injection force increases linearly with the injection flow rate and decreases with the use of either a smaller-gauge needle (larger inner diameter) or a thinner-walled needle (Figure 10, panels b-d, and Figure 11, panel a). Using a clinically relevant 27-gauge UTW needle and a 1 mL / min flow rate, the 460 mg / mL BSA suspension had an injection force of only 14 N (Figure 11, panel a), well within the injection force range associated with pen-type autoinjectors (see, e.g., Hill, R. Lett. et al. Med Devices-Evid Res 9, 257-266 (2016)). Long-term storage experiments further demonstrate that the suspension exhibits minimal particle settling after 35 days of storage in the triacetin nonsolvent (Figure 11, panel b) and at comparable injection forces (21.4 N vs. 22.4 N) (Figure 11, panel c).

[0249] 6.1.6. Example 6 - Evaluation of stability after storage and stress aging. Exemplary BSA protein particle compositions were evaluated for storage stability.

[0250] Figure 12, panels a-b. (Panel a) Further illustrating the SEC traces of 20 mg / mL BSA in PBS, 460 mg / mL spray-dried BSA without MoNi in triacetin, and 460 mg / mL spray-dried BSA with 5 wt% MoNi in triacetin after stress aging at 60°C for 30 minutes. (Panel b) The corresponding monomer fraction of each formulation after stress aging.

[0251] As described above, the comparative stability of BSA in solution and in triacetin suspension was evaluated via a stress aging assay. BSA protein suspensions were prepared at 460 mg / mL in triacetin using spray-dried BSA particles with 5 wt% MoNi and BSA particles without 5 wt% MoNi. Aqueous BSA controls were prepared by dissolving fresh lyophilized BSA in PBS at 20 mg / mL. The samples were heated at 60°C for 30 minutes, and their stability was then evaluated by SEC (Figure 12, panel a). BSA dissolved in PBS showed significant aggregation and a reduced monomer peak fraction (37%), whereas solid BSA particles dispersed in a nonsolvent remained stable (>80% monomer peak fraction) throughout the elevated temperature conditions (Figure 12, panel b). To further compare the stability of BSA suspensions with and without MoNi, 460 mg / mL suspensions in triacetin were stored at 4° C., 25° C., and 37° C. for 48 and 120 hours, and stability was assessed by SEC. The suspensions with MoNi showed a smaller high molecular weight aggregate peak than the suspensions with BSA alone (see Example 7).

[0252] 6.1.7. Example 7 - Protein stability during storage in suspension form over time. To investigate the stability of the protein in the injectable formulation during storage over time, Formulation 11 (containing spray-dried particles with BSA and copolymer MoNi in triacetin at a total protein concentration of 460 mg / mL) was stored at various temperatures (4°C, 25°C, and 37°C) and samples were evaluated after 48 and 120 hours.

[0253] A BSA protein suspension with 5 wt% MoNi was prepared at 460 mg / mL in triacetin. The protein suspension was loaded into a 1 mL Thermo Fisher Luer slip syringe and capped with a BD26G 1 / 2 inch needle wrapped in parafilm to limit solvent evaporation. The injection force of the 460 mg / mL protein suspension was measured on day 0 and again on day 35. The syringe was stored horizontally at 23°C.

[0254] Figure 13, panels a(i)-a(iii). Stability of 460 mg / mL BSA formulations in triacetin with and without 5 wt% MoNi was evaluated by storage at 4°C, 25°C, and 37°C for 48 hours. Panel a(i): Full SEC traces of the formulations after 48 hours. Panel a(ii): Dimer peaks for all formulations after 48 hours. Panel a(iii): High molecular weight peaks for all formulations after 48 hours. Overall, all formulations showed good stability after 48 hours, with the majority of the light scattering signal coming from the monomer peak. On average, samples containing MoNi showed smaller dimer and high molecular weight peaks than samples without MoNi, suggesting improved storage stability. SEC traces were obtained using a Superose 6 column. PBS with sodium azide was used as the eluent.

[0255] Figure 14, panels a(i)-a(iii). Stability of 460 mg / mL BSA formulations in triacetin with and without 5 wt% MoNi was evaluated by storage at 4°C, 25°C, and 37°C for 120 hours. Panel a(i): Full SEC traces of the formulations after 120 hours. Panel a(ii): Dimer peak for all formulations after 120 hours. Panel a(iii): High molecular weight peak for all formulations after 120 hours. BSA formulations containing MoNi stored in syringes for 120 hours show no particle settling. Overall, all formulations show good stability after 120 hours, with the majority of the light scattering signal coming from the monomer peak. On average, samples containing MoNi show a smaller high molecular weight peak than samples without MoNi, suggesting improved storage stability. SEC traces were obtained using a Superose 6 column. PBS with sodium azide was used as the eluent.

[0256] 6.1.8. Example 8 - Liquid Carrier Studies. To investigate the liquid carrier component of the subject composition, several solvents were tested for their suspension performance characteristics. The compositions were prepared by dispersing BSA / MoNi (5%) microparticles in each liquid carrier at 460 mg / mL. The criteria for selecting a suitable liquid carrier include: i) a viscosity of less than 25 cp at 25°C; ii) whether a particular solvent has previously been used in FDA-approved parenteral drug products at exposure levels above those targeted in this disclosure; iii) dispersion time upon administration, i.e., at a rate that ensures that the formulation does not form a depot; iv) compatibility with the contact surface, i.e., not dissolving the silicon glass treatment; v) insufficient protein solubilization, i.e., ensuring it is a non-solvent for the particles; vi) having a density that closely matches that of the particle density, i.e., maximizing suspension stability; and vii) Low volatility, noting that volatility concerns can be addressed by combining higher and lower volatility solvents and by using airtight storage conditions.

[0257] Table 4 illustrates the various liquid carriers that were tested along with a rating of their suspension performance. Suspension Performance Legend:

number

[0258] media density research As noted above, one of the criteria for selecting a suitable liquid carrier is that the density of the liquid carrier be as close as possible to the density of the particles in the composition so as to maximize suspension stability.

[0259] Particle sedimentation can be experimentally evaluated through centrifuge studies. In such studies, BSA is added at a low concentration (50 mg / mL) to various non-solvent mixtures. The resulting mixtures are then centrifuged for equal periods of time, after which the volume of BSA powder that settles to the base of each formulation is compared. If the non-solvent mixture and BSA powder are more similar in density, reduced sedimentation is observed.

[0260] FIG. 15 shows the results of a centrifuge study to evaluate suspension stability in the following vehicles, from left to right: triacetin alone, triacetin with 20% benzyl benzoate, triacetin with 20% benzyl alcohol, and triacetin with 20% safflower oil.

[0261] Media viscosity study As mentioned above, one of the criteria for selecting a suitable liquid carrier is that it have a low viscosity (ie, a viscosity of less than 25 cp at 25°C).

[0262] In order to improve the injectability of suspension, it may be advantageous to reduce the viscosity of the liquid carrier by using a combination of solvents in the liquid carrier.By adding a low-viscosity non-solvent to a higher-viscosity non-solvent, a decrease in viscosity has been observed.Because this viscosity decrease is non-linear, even adding a small amount of a low-viscosity non-solvent may improve viscosity and the injectability of suspension.

[0263] Figure 16 illustrates viscometer measurements of various liquid carriers. This graph illustrates how the viscosity of a higher viscosity non-solvent (propylene glycol (PG) or triacetin (T)) can be reduced by adding a lower viscosity additive (such as BA).

[0264] This experiment demonstrates that BA can significantly reduce the viscosity of a solvent mixture containing PG and T. This technique can be used to understand the viscosity of other non-solvent blends.

[0265] Injection force experiment Injection force experiments were performed on 450 mg / mL BSA suspensions in triacetin:DMAc and triacetin:DMAc:BA blends. Injections were performed at 1 mL / min through a 26G 1 / 2 inch needle.

[0266] Figure 17, panels a-d. Injection force measurements (1 mL / min through a 26 G, ½ inch needle) show a significant decrease in injection force when DMAc is used in combination with triacetin as a non-solvent. Panel a shows the plateau injection force of a triacetin suspension (100%) versus suspensions in various blends of triacetin:DMAc and triacetin:DMAc:BA. Panel b shows the injection force of a triacetin suspension versus DMAc content. Panel c shows the injection force over time of a triacetin suspension (100%) versus suspensions in various blends of triacetin:DMAc and triacetin:DMAc:BA. Panel d shows that the addition of polymeric MoNi affects the consistency of BSA powder dispersed in DMAc. The left image in panel d illustrates a formulation with 450 mg / mL BSA DMAc with MoNi. The right image in panel d illustrates a formulation with 450 mg / mL BSA in DMAc without the addition of MoNi.

[0267] This data shows that adding a lower viscosity non-solvent is also advantageous for reducing injection force. Here, a 2.5-fold reduction in injection force is observed when DMAc is used in a 75:25 v / v ratio compared to triacetin alone. When DMAc is used alone, a 7-fold reduction in injection force is observed compared to triacetin. This injection force measurement technique can be used to understand the injectability of other non-solvent blends. A blend containing triacetin:DMAc:BA (70:20:10) also demonstrates reduced injection force.

[0268] 6.1.9. Example 9 - Tolerance of Low Viscosity Non-Solvent Additives Tolerability screening was used to quantify the tolerability of injecting low-viscosity nonsolvent excipients including benzoyl alcohol (BA), NMP, and DMAc.

[0269] SKH1-E mice were subcutaneously administered 10 μL of vehicle or a vehicle blend with triacetin (protein-free). Over the next 24 hours, the extent of skin reaction to the subcutaneously injected vehicle was quantified. The volume of vehicle injected was twice the volume of vehicle administered to mice as a paste.

[0270] The grading scale in Table 5 was used to quantify the severity of adverse skin reactions. [Table 5]

[0271] Tolerability screening results are shown in Table 6. [Table 6]

[0272] Tolerance screening results demonstrated that triacetin was well tolerated. Mice administered triacetin alone did not exhibit skin swelling or redness. While DMAc or BA alone caused skin irritation, blends with less than 50:50 v / v DMAc or BA in triacetin were also well tolerated. These blends demonstrated an adverse skin reaction grade of 1 or less in the first 24 hours after subcutaneous injection. Subcutaneous injection of NMP caused greater skin irritation than DMAc or BA.

[0273] Tolerance studies demonstrate that triacetin, DMAc, and BA are promising nonsolvents for in vivo use. Initial in vivo work focused on triacetin and DMAc because of the improved slurry stability and lower injection force of this blend.

[0274] 6.1.10. Example 10 - In vivo experiments demonstrating subcutaneous administration of BSA protein slurry Fluorescently labeled BSA particles were obtained by spray-drying AF647-BSA with unlabeled BSA at a ratio of 1:500. MoNi was maintained at 5 wt% in the final particle formulation. The fluorescently labeled BSA particles were then further diluted with unlabeled BSA particles with MoNi at a ratio of 1:5, resulting in a final ratio of AF647-BSA:unlabeled BSA of 1:2500. Protein suspensions in triacetin and 70 triacetin:30 DMAc were formulated as previously described herein. The bolus control consisted of BSA dissolved in 20 mg / mL PBS at a ratio of 1:2500 AF647-BSA:unlabeled BSA.

[0275] A) Protein slurry in triacetin medium SKH1-E mice were subcutaneously administered either 200 μL of 20 mg / mL BSA dissolved in PBS (bolus injection) or 9 μL of 450 mg / mL BSA in triacetin slurry (high-concentration paste or high-concentration protein suspension) (4 mg total BSA in both groups). The BSA slurry was administered using a Hamilton syringe and a 27G 1 / 2-inch needle. The BSA administered to the animals was labeled with Alexa647 dye, which allows for fluorescent imaging of protein remaining in the subcutaneous space at an excitation wavelength of 600 nm and an emission wavelength of 670 nm. The total fluorescence intensity corresponding to each animal was plotted over time over a period ranging from 1 h to 24 h and fitted to a single-phase exponential decay model. The half-life of subcutaneous absorption was obtained and averaged using GraphPad Prism. ------------- 1 BSA concentrations assume a particle density of 1 g / cm^3. See Example 11 below for BSA particle density. BSA concentrations are estimated to be close to 506 mg / mL.

[0276] B) Protein slurry in 70:30 triacetin:DMAc medium SKH1-Elite mice were each subcutaneously injected with either 8.8 μL of a 450 mg / mL fluorescently labeled BSA protein suspension (in triacetin or 70% triacetin:30% DMAc) or 200 μL of 20 mg / mL fluorescently labeled BSA in PBS. Protein suspension injections were administered using a 50 μL Hamilton syringe with a 26G 1 / 2-inch needle. Bolus injections were administered using a 1 mL Luer-Lok syringe with a 26G 1 / 2-inch needle. The animals' subcutaneous injection sites were imaged using IVIS (Lago) over a series of time points over a 2-day period.

[0277] During imaging, mice were anesthetized with isoflurane gas and imaged at an exposure time of 2 seconds, excitation wavelength of 600 nm, and emission wavelength of 670 nm (medium binning; F / stop, 1.2). Total radiant efficiency [(photons / s) / (µW / cm2)] was quantified using an equally sized region of interest surrounding the injection site. Because early time points showed increasing, rather than decreasing, fluorescence in the region of interest due to quenching effects, the fluorescence intensity at each time point was normalized to the fluorescence intensity at the time of the first fluorescence decrease. Normalized fluorescence intensity values ​​for each mouse (n = 3–5) were fitted to a single exponential decay model, and half-lives were obtained and averaged using GraphPad Prism.

[0278] Panels a-c of Figure 18 and panels a-b of Figure 19 demonstrate that the subject high concentration BSA suspensions enable in vivo administration and subcutaneous absorption.

[0279] Figure 18, panels a-c, show the results of in vivo administration of 4 mg of BSA to mice via either a 200 μL injection of 20 mg / mL BSA in PBS (bolus injection) or a 9 μL injection of 450 mg / mL BSA in triacetin (high-concentration paste / high-concentration protein suspension) (N≧3 per group). (Panel a) Representative IVIS images demonstrating subcutaneous absorption of fluorescently labeled BSA administered via PBS bolus injection or high-concentration protein suspension. (Panel b) Comparative volumes of administration of bolus injection and high-concentration protein suspension. (Panel c) Half-life of subcutaneous absorption of high-concentration paste and bolus injection.

[0280] The results show that despite the large difference in injection volume required, paste and bolus injections exhibit comparable subcutaneous absorption kinetics with a mean absorption half-life of 4 hours (Figure 18, panel c, N > 3 per group). It is interesting to note that animals receiving PBS bolus injections appeared to have more residual fluorescence at later time points (Figure 18, panel a). This may indicate decreased overall absorption when larger injection volumes are given.

[0281] Figure 19, panels a-b, show the results of in vivo administration of 9 uL of 450 mg / mL BSA in a 70:30 triacetin:DMAc slurry. Panel a shows the fluorescence signal in the subcutaneous space, which fits a monophasic exponential decay mode, identifying the half-life of subcutaneous absorption. Panel b shows the comparative half-life of subcutaneous absorption of BSA administered via PBS bolus injection or a protein suspension formulated in triacetin or 70 triacetin:30 DMAc.

[0282] Results showed comparable subcutaneous absorption kinetics between the 70:30 triacetin:DMAc slurry and the 100% triacetin slurry, with a mean absorption half-life of 4 hours, similar to that of a PBS bolus injection (20 mg / mL BSA in PBS).

[0283] 6.1.11. Example 11 - Measurement of BSA Particle Density The particle density of spray-dried particles can be measured using a Micromeritics pycnometer. The powder mass is measured at the beginning of the run, and the pycnometer measures the volume of the cells. Pycnometer measurements of the BSA powder with MoNi yield a powder mass of 158.68 mg and a powder volume of 0.1208 cm^3 (SD: 0.002 cm^3). This is an estimated density of 1.31 g / cm^3. Density results are consistent across multiple powder batches and multiple runs.

[0284] All reported protein concentrations discussed herein assume a spray-dried particle density of 1. Because the powder incorporated into the suspension was measured by mass and assumed to take up a volume based on a density of 1 g / cm^3, a higher spray-dried particle density means that the disclosed pastes likely have higher protein concentrations than originally estimated. Table 7 shows the recalculation of protein concentrations. [Table 7]

[0285] Figure 24, panels a-b: Panel a depicts the comparative protein concentration predicted from density measurements and measured via Nanodrop from known slurry volumes. Panel b shows the injection force as a function of concentration of BSA particles (with mole % matched MoNi or Tween 80) in 70 triacetin:30 DMAc. The injection force as a function of particle concentration is fitted to a particle jamming model. The left column of Figure 24, panel a, is generated by adjusting the calculation formula based on a density of 1.31 g / cm^3, while the right column is generated by taking a known volume of slurry, dissolving it in water, and measuring the actual protein content with UV-vis spectroscopy.

[0286] BSA particles spray dried with an equimolar ratio of Tween 80 exhibit similar size and surface morphology (see Figure 23, panel c), but exhibit a higher injection force at equal BSA concentrations, as seen in Figure 24, panel b. Figure 24, panel b. Injection force measurements with BSA-MoNi in 70 triacetin:30 DMAc demonstrate injection of 600 mg / mL BSA with a clinically relevant injection force of 17 N through a 26 G ½ inch needle at 1 mL / min. The concentration versus injection force data were fitted to a particle jamming model (Mueller, S. et al., P Roy Soc a-Math Phys 466, 1201-1228 (2010); Krieger, I. M. et al. T Soc Rheol 3, 137-152 (1959)) and, using the sphericity coefficient, it was found that the BSA-MoNi particles achieved near the theoretical maximum packing density of spheres (volume fraction = 0.74) before jamming (Figure 24, panel b), and a lower maximum injectable concentration upon particle jamming (Figure 24, panel b, and Figure 23, panels a-b).

[0287] 6.1.12. Example 12 - Comparison with Polysorbate and Pluronic Controls Polysorbates and Pluronics are nonionic surfactants that can be added to the spray-drying process to reduce surface tension and improve protein stability. Polysorbate 80 (Tween 80) and Pluronic L-61 were selected as controls. These Pluronic and polysorbate controls were formulated to match the molar ratio of MoNi used in the current formulation. Table 8 summarizes the formulations containing each additive. [Table 8]

[0288] Without being bound by any particular theory, it is hypothesized that MoNi may be a preferred spray drying additive over Polysorbate 80 or Pluronic L-61 due to its higher glass transition temperature. MoNi has a glass transition temperature of 135°C, while Tween 80 and Pluronic L-61 have glass transition temperatures much lower than room temperature. The higher glass transition temperature may result in improved stability throughout the spray drying process and a less sticky product, as well as higher yields.

[0289] Powder quality and yield Figure 20, panels a-b, illustrate the products obtained after spray-drying formulations containing polysorbate 80 and Pluronic L-61. Panel a shows images of Falcon tubes containing comparative products recovered from spray-drying equal amounts of BSA with polysorbate 80 (left tube) and Pluronic L-61 (right tube). Panel b shows the spray-dryer glass components after spray-drying Pluronic L-61. As can be seen in the images, spray-drying Pluronic L-61 results in a sticky powder that adheres to the spray-dryer glass components, thus resulting in a low yield of spray-dried product (see panel a, right tube).

[0290] Powder stability throughout the spray drying process Figure 21, panels a-c, illustrate SEC traces of fresh BSA and BSA spray-dried with MoNi (BSA_MoNi), polysorbate 80 (BSA_Tw80), and without any additives (BSA_NoMoNi) (fresh BSA standard). (Samples were run on a Superose6 10 / 300GL column at 0.5 mL / min.) Panel a shows that for all spray-dried samples, the monomer and dimer peaks appear similar. Panel b shows the difference between the spray-dried samples for the high molecular weight aggregate peak. Panel c shows that the area fraction of the high molecular weight peak is smallest for the fresh BSA standard, followed by BSA spray-dried in MoNi, BSA spray-dried with polysorbate 80, and finally, BSA spray-dried without any additives.

[0291] The SEC traces suggest that MoNi is beneficial in preserving protein stability during the spray-drying process. Samples spray-dried with MoNi exhibited smaller high molecular weight peaks (likely corresponding to aggregates) than samples spray-dried either with polysorbate 80 or without the additive. While BSA is a very stable protein, a more pronounced effect would be expected if the stability of proteins that are more difficult to spray-dry, such as monoclonal antibodies, were examined.

[0292] Comparison of stress aging of powders formulated with MoNi and polysorbate 80 To understand the stability of protein pastes made with BSA, BSA made with MoNi, and BSA made with polysorbate 80, pastes of 450 mg / mL BSA in triacetin were aged for 60 minutes at 60°C. 20 mg / mL BSA in PBS aged under the same conditions was used as a positive control.

[0293] After stress aging, the samples were run on SEC to look for changes in the area fraction of the high molecular weight aggregate peak.

[0294] Figure 22, panels a-b, illustrate the full SEC trace and the high molecular weight aggregate peak trace for aged protein pastes made with BSA, BSA with MoNi, and BSA with polysorbate 80. BSA aged at 20 mg / mL in PBS was used as a control. All samples were aged at 60°C for 60 minutes. Samples were run on a Superose6 10 / 300GL column at 0.5 mL / min. Table 9 shows the sample legend for Figure 22. Panel a shows the difference between samples for the high molecular weight aggregate peak. Panel b shows the area percentage of the high molecular weight peak for each sample. [Table 9]

[0295] The SEC traces suggest that MoNi is beneficial in maintaining protein stability during the aging process. Aged samples containing MoNi exhibited smaller high molecular weight peaks than samples containing polysorbate 80 or no additive. The 20 mg / mL control in PBS exhibited significant aggregation during the stress aging experiment.

[0296] Comparison of the injectability of powders formulated with MoNi and polysorbate 80 Figure 23, panels a-c, show injection force experiments (1 mL / min through a 26 G ½ inch needle) using a 450 mg / mL BSA suspension in triacetin. BSA was spray-dried with either polysorbate 80 (Tween 80) or MoNi in equimolar amounts (7.14 μmol per gram of BSA). Panel a shows the injection force (N) over time (seconds). Panel b shows the plateau injection force for formulations containing MoNi versus polysorbate 80 (Tween 80). Panel c illustrates SEM of the particle morphology of i) BSA MoNi and ii) BSA polysorbate 80 (Tween 80) particles (scale bar 10 μm).

[0297] The results show that the paste formulated with polysorbate 80 exhibits a 43% increase in injectability when formulated at an equimolar ratio of MoNi and polysorbate 80 (7.14 μmol per gram of BSA). It is relevant to note that MoNi is much less toxic than polysorbate 80, allowing for a larger formulation design space.

[0298] 6.1.13. Example 13 - Bovine Serum Albumin (BSA) Microparticle Suspension Aqueous formulations of bovine serum albumin (BSA) were prepared and spray dried under the conditions in Table 10 to provide BSA microparticle formulations. [Table 10]

[0299] Microparticle formulations 004A and 004B were imaged by scanning electron microscopy (SEM) and both formulations were observed to exhibit smooth particles below 5 μm.

[0300] Figure 25, panels a-b, show SEM images of BSA microparticle formulations. Panel a shows an SEM image of formulation 004A (smooth particles < 5 μm). Panel b shows an SEM image of formulation 004B (smooth particles < 5 μm).

[0301] Formulations 004A and 004B were suspended in triacetin and loaded into 1 mL Schott syringes (27G thin-walled needles and V9519 coated plungers). Both formulations showed consistent suspension with no particle settling or phase separation observed.

[0302] Figure 26, panels a-b, show photographic images of BSA microparticle formulations suspended in triacetin and loaded into 1 mL Schott syringes. Panel a shows a photographic image of a syringe loaded with formulation 004A (480 mg / mL) suspended in triacetin. Panel b shows a photographic image of a syringe loaded with formulation 004B (458 mg / mL) suspended in triacetin.

[0303] To investigate the injectability of BSA microparticle formulations, injection force measurements were performed on suspensions of formulations 004A and 004B in triacetin injected at different flow rates from a 1 mL Schott syringe (27 G thin-walled needle and V9519 coated plunger).

[0304] Figure 27, panels a-b, are graphs of the injection force (N) of BSA microparticle suspensions over 5 seconds at various flow rates. Panel a shows the injection force (N) over 5 seconds of BSA microparticle suspension 004A (480 mg / mL BSA) in triacetin injected at 2 mL / min, 4 mL / min, and 6 mL / min from a 1 mL Schott syringe (27 G thin-wall needle and V9519-coated plunger). Panel b shows the injection force (N) over 5 seconds of BSA microparticle suspension 004B (458 mg / mL BSA) in triacetin injected at 2 mL / min, 4 mL / min, and 6 mL / min from a 1 mL Schott syringe (27 G thin-wall needle and V9519-coated plunger).

[0305] Figure 28 illustrates injection force measurements of 004A (480 mg / mL BSA) and 004B (458 mg / mL) microparticle suspensions in triacetin injected at different flow rates from a 1 mL Schott syringe (27G thin-walled needle and V9519 coated plunger). The graph shows that the injection force (N) of suspensions of 004A and 004B in triacetin is linear with flow rate.

[0306] 6.1.14. Example 14 - In vivo experiments demonstrating subcutaneous administration of human immunoglobulin G (hIgG) protein slurry Following the success with BSA, in vivo experiments were applied to human immunoglobulin G (hIgG), which is noteworthy because it is directly comparable to mAb therapeutics in both chemical structure, solubility, and size.

[0307] Human immunoglobulin G (hIgG) was fluorescently labeled with Alexa-647-NHS from lumiprobe at a 5:5 weight ratio. Briefly, 50 mg of dry hIgG was dissolved in 10 mL of PBS, and 500 μL of a 5 mg / mL DMSO stock solution of Alexa-647-NHS was added to the solution. The reaction proceeded for 24 hours at room temperature, and free dye was removed with a 10 kDa MWCO Amicon spin filter.

[0308] Fluorescently labeled hIgG particles were obtained by spray-drying Alexa Fluor 647 dye (AF647)-labeled hIgG with unlabeled BSA at a ratio of 1:20. MoNi was maintained at 5 wt% in the final particle formulation. The fluorescently labeled hIgG particles were then further diluted with unlabeled hIgG particles with MoNi at a ratio of 1:100, resulting in a final ratio of AF647-labeled hIgG:unlabeled hIgG of 1:2000. A protein suspension in triacetin was formulated as described above. The bolus control consisted of hIgG dissolved in 100 mg / mL PBS at a ratio of 1:2000 AF647-labeled hIgG:unlabeled hIgG.

[0309] SKH1-Elite mice were subcutaneously injected with either 20 μL of a 300 mg / mL fluorescently labeled hIgG protein suspension in triacetin or 60 μL of a 100 mg / mL fluorescently labeled hIgG in PBS. Both the protein suspension and the bolus injection were administered with an insulin syringe equipped with a 28-gauge needle. The animals' subcutaneous injection sites were imaged using IVIS (Lago) over a series of time points over a two-day period.

[0310] During imaging, mice were anesthetized with isoflurane gas and imaged at an exposure time of 1 s, excitation wavelength of 600 nm, and emission wavelength of 670 nm (medium binning; F / stop, 1.2). Total radiant efficiency [(photons / s) / (µW / cm2)] was quantified using an equally sized region of interest surrounding the injection site. Because early time points showed increasing, rather than decreasing, fluorescence in the region of interest due to quenching effects, the fluorescence intensity at each time point was normalized to the fluorescence intensity at the time of the first fluorescence decrease. Normalized fluorescence intensity values ​​for each mouse (n = 3–5) were fitted to a single exponential decay model, and half-lives were obtained and averaged using GraphPad Prism.

[0311] Figure 29, panels a-g, illustrates the effective use of high-concentration suspension technology to deliver hIgG. Panel a depicts an SEM of spray-dried IgG with MoNi particle morphology (scale bar is 10 μm). Panel b depicts SEC traces of a fresh hIgG control, spray-dried hIgG without MoNi, spray-dried hIgG with 5 wt% MoNi, and spray-dried hIgG with 25 wt% trehalose and 5 wt% MoNi. PBS with sodium azide is used as the eluent. Panel c shows the injection force as a function of concentration of hIgG microparticles with MoNi in triacetin. The injection force as a function of particle concentration is fitted to a particle jamming model. (Panel d) Injection force for a 1 mL / min injection of a 350 mg / mL hIgG, 5 wt% MoNi suspension through a 26G ½ inch needle with DMAc, benzyl alcohol (BA), and benzyl benzoate (BB) non-solvent additives. (Panel ei) Representative IVIS images demonstrating subcutaneous absorption of fluorescently labeled hIgG administered via PBS bolus injection or high-concentration protein suspension. (Panel e.ii) The fluorescent signal in the subcutaneous space was fitted to a monophasic exponential decay mode to identify the half-life of subcutaneous absorption. (Panel f) Comparative half-life of subcutaneous absorption of hIgG administered via PBS bolus injection or UHC protein suspension formulated in triacetin. (Panel g) Comparative volumes of administration of bolus injection and high-concentration protein suspension.

[0312] Similar to the BSA formulation described in Example 10, the hIgG to MoNi ratio in the final spray-dried product was fixed at 20:1, and the initial feedstock concentration and spray-drying parameters were optimized to produce spherical particles. As shown in Figure 29, panel a, spray-dried hIgG particles with 5 wt% MoNi are 5-20 μm in diameter and have a spherical morphology. As previously observed with BSA, MoNi stabilized hIgG during the high-temperature spray-drying process. SEC characterization of fresh hIgG and spray-dried hIgG with and without MoNi indicates that spray-drying hIgG with MoNi results in a higher monomer peak fraction and reduced high-molecular-weight aggregates (Figure 29, panel b). Importantly, a control hIgG formulation utilizing 30 wt% trehalose as a stabilizer was found to have a similar aggregate fraction as hIgG with 5 wt% MoNi, and the addition of 25 wt% trehalose to a 5 wt% MoNi formulation did not provide any additional stability benefit to the spray-dried product (Figure 30). In summary, these results indicate that only 5 wt% MoNi was required to stabilize hIgG during the spray-drying process, compared to state-of-the-art formulations containing a minimum of 30 wt% trehalose (Maury, M. et al. Eur J Pharm Biopharm 59, 251-261 (2005)).

[0313] With these high hIgG-containing microparticles (95% by weight), a suspension was formulated using triacetin as a non-solvent, and the concentration was 1 g / cm 3Assuming a particle density of 0.01, we were able to measure the injection force as a function of hIgG concentration. Injection force measurements with hIgG-MoNi in triacetin demonstrate injection of 350 mg / mL hIgG with an injection force of 22 N at 1 mL / min through a 26 G 1 / 2-inch needle (Figure 29, panel c). To understand particle interactions, the concentration vs. injection force data were again fitted to a particle jamming model using a spherical shape factor (Mueller, S. et al., P Roy Soca-Math Phys 466, 1201-1228 (2010)) (Figure 29, panel c). Considering that hIgG-MoNi particles have a similar morphology to BSA-MoNi particles, as determined by SEM imaging, jamming at this low concentration implies greater particle-particle interactions, which is likely due to the physical differences between dried hydrophobic antibody versus dried hydrophilic albumin. As seen with BSA, the injection force of hIgG suspensions can be further reduced by changing the non-solvent composition. 70 triacetin:30 DMAc and 70 triacetin:30 BA reduce the injection force of a 350 mg / mL hIgG suspension (22 N) by 2.75-fold (8 N) and 1.5-fold (15 N), respectively. Additionally, benzyl benzoate, a low-viscosity FDA-approved excipient often used as a preservative, reduces the injection force by 1.8-fold (12 N) (Figure 29, panel d).

[0314] To demonstrate the in vivo delivery and subcutaneous absorption of hIgG-MoNi suspension as described above, SKH1e mice were subcutaneously injected through a 28-gauge insulin needle with an equal dose of fluorescently labeled hIgG in either a 300 mg / mL triacetin suspension or a 100 mg / mL PBS bolus (Figure 31 shows the injection force curve for injection of a 300 mg / mL IgG, 5 wt% MoNi suspension at 1 mL / min through a BD insulin syringe). The higher-concentration IgG bolus was selected as representative of a higher-concentration aqueous mAb product. Fluorescence images collected from an in vivo imaging system (IVIS) were then utilized to study the subcutaneous absorption of hIgG at the injection site (Figure 29, panels e.i.). The fluorescence signal over time from the injected protein suspension and bolus was normalized and fitted to a monophasic exponential decay curve to provide the half-life of subcutaneous hIgG absorption (Figure 29, panels e.ii.). The half-life of subcutaneous hIgG absorption was approximately 9 hours (Figure 29, panel f), consistent with previous literature values ​​(Deng, R. et al. Subcutaneous bioavailability of therapeutic antibodies as a function of FcRn binding affinity in mice. MAbs 4, 101-109 (2012)). Despite a three-fold difference in required injection volume, the difference in absorption half-life between the administration methods was not statistically significant (Figure 29, panel g).

[0315] 6.1.15. Example 15 - Co-formulation of insulin and pramlintide A storage stable co-formulation of insulin and pramlintide that is injectable through clinically relevant needle gauges was investigated.

[0316] Spray drying of insulin and pramlintide particles: A feed solution for spray-dried insulin particles was formed by diluting U500 insulin stock with cell-grade water containing trehalose and MoNi. The insulin feed solution contained insulin (0.99 mg / mL), glycerin (0.91 mg / mL), metacresol (0.14 mg / mL), zinc (0.005 mg / mL), MoNi (2.5 mg / mL), and trehalose (45.4 mg / mL) (50 mg / mL total). A feed solution for spray-dried pramlintide particles was formed by diluting 5 mg / mL pramlintide stock with cell-grade water containing trehalose and MoNi. The pramlintide stock solution contained pramlintide (0.23 mg / mL), MoNi (2.5 mg / mL), and trehalose (47.3 mg / mL) (50 mg / mL total).

[0317] The solution was spray dried at a concentration of 50 mg / mL solids. The solution was formulated in the lab and transferred to the spray dryer on ice. The solution was allowed to warm to room temperature before being spray dried. The total time between preparation of the feed solution and spray drying was approximately 4 hours.

[0318] The particles were spray dried using a Buchi B-290 spray dryer equipped with a high-efficiency cyclone. The spray dryer parameters were: inlet temperature: 130 °C, outlet temperature: 78 °C, 5% pump (2–3 mL / min), 40 mm air, nozzle wash: 3, room temperature: 20 °C, and room humidity: 42%.

[0319] Figure 32, panels a-b, show SEC traces of insulin before and after spray drying. (Panel a) shows the insulin SEC traces of spray-dried and pre-spray-dried insulin over a 22-minute period. (Panel b) is a zoomed-in portion of the SEC trace in panel a from 14 to 17 minutes.

[0320] Figure 33, panels a-b, show SEC traces of pramlintide before and after spray drying. (Panel a) shows the SEC traces of spray-dried and pre-spray-dried pramlintide over a 20-minute period. (Panel b) is a zoomed-in portion of the SEC trace in panel a from 14 to 17 minutes.

[0321] FIG. 34 shows SEC traces of pramlintide (“pram”), insulin, and a control before and after spray drying.

[0322] Although the SEC curves of spray-dried insulin and pramlintide initially appear to have a high molecular weight tail, further characterization demonstrates that the high molecular weight peak can be attributed to the high concentration of MoNi in the spray-dried powder samples, rather than protein aggregation. Samples containing MoNi, trehalose, and pramlintide before and after spray drying show identical SEC traces (Figure 34). The high molecular weight MoNi peak in the spray-dried insulin sample appears smaller than the peak in the pramlintide sample because the SEC sample (formulated with 0.66 mg / mL of therapeutic protein) contains only 1.75 mg / mL of MoNi, compared to the pramlintide SEC sample, which contains 7.53 mg / mL of MoNi.

[0323] Resuspension of insulin and pramlintide particles in triacetin: Insulin and pramlintide spray-dried particles were resuspended in triacetin. The particles were left in triacetin for 24 hours before being imaged. As expected from conventional SEM imaging, the particles appear to be 5-20 μm in diameter, with most particles being less than 10 μm (Figure 35).

[0324] Figure 35, panels a-b. SEM images of insulin and pramlintide spray-dried particles resuspended in triacetin (panel a), and trehalose and MoNi (95:5) particles (panel b). The particles in panel b contain neither insulin nor pramlintide, so proteins may affect particle morphology.

[0325] Figure 36 shows insulin and pramlintide spray-dried particles resuspended in triacetin at a solids content of 350 mg / mL. The particle suspension flows like a liquid.

[0326] The co-formulation of injectable insulin and pramlintide has a solids content of 350 mg / mL. It contains 175 mg / mL of insulin powder particles and 175 mg / mL of pramlintide powder particles resuspended in triacetin. The final composition yields a standard insulin dose of 3.47 mg / mL (100 insulin units per mL) and a pramlintide dose of 0.8 mg / mL.

[0327] Figure 37, panels a and b, shows the injection force of insulin pramlintide co-formulation in triacetin (350 mg / mL solids). Injection measurements were performed using a 1 mL syringe at 1 mL / min through a 26G ½ inch needle.

[0328] The current co-formulation was easily injectable through clinically relevant needle gauges, and no clogging was observed. It may be relevant to increase the overall solids content of the suspension to prevent particle settling in the final formulation, especially if the formulation is stored in a syringe prior to injection.

[0329] Redissolution of the co-formulation in water: Water was added to the co-formulation paste and the formulation dissolved clearly with little visible clumping.

[0330] FIG. 38 is an image depicting insulin pramlintide co-formulation reconstituted in cell grade water.

[0331] 6.1.16. Example 16 - Syringe Force Measurements for Various Liquid Carriers To investigate the syringe force profiles (e.g., breakloose (initiation) force, and glide (sustained) force) of various liquid carriers in syringes, the following vehicle-only syringes were loaded in duplicate (n=2): 1. Triacetin (100%) 2. Benzyl benzoate (100%) 3. Miglyol 810 (100%) 4. Miglyol 840 (100%) 5. Benzyl alcohol (100%) 6. Ethyl oleate (100%).

[0332] The syringes were stored at ambient temperature for 48 hours, protected from light. No visual changes were observed when comparing the appearance of each upon loading with the same appearance 48 hours after storage.

[0333] Syringe force measurements (n=2) were performed on each vehicle after 48 hours, and the results are shown in Table 11. [Table 11]

[0334] Overall, comparison of syringe force profiles (lowest to highest): Benzyl alcohol ~ Ethyl oleate < Benzyl benzoate ~ Miglyol 840 < Triacetin ~ Miglyol 810.

[0335] 6.1.17. Example 17 - BSA and MoNi Suspension in Prefilled Syringes Syringe force profiles of injectable formulations containing BSA and MoNi suspensions (460 mg / mL) in various liquid carriers. Exemplary formulations were prepared according to General Method B (described herein). Table 12 below summarizes the formulations prepared for this example. [Table 12]

[0336] Formulations 12, 13, 14, and 16 were prepared as described above, loaded into syringes in duplicate (n=2), and stored at ambient temperature for 48 hours, protected from light. The appearance of each formulation at 48 hours was compared with the appearance at the time of loading. After 48 hours of storage in prefilled syringes, the following observations were made: Formulation 12 (benzyl benzoate): no visual change, Formulation 13 (Miglyol 840): no visual changes, Formulation 14 (benzyl alcohol): The suspension became a solid material, and Formulation 16 (ethyl oleate): No visual change.

[0337] In summary, no changes were observed in formulations 12, 13, and 16, which contain benzyl benzoate, Miglyol 840, and ethyl oleate as liquid carriers, respectively, after 48 hours of storage in prefilled syringes. Thus, formulations 12, 13, and 16 were loaded into prefilled syringes and further evaluated to establish syringe force profiles after 48 hours of storage. Prefilled syringes were subjected to force measurements in duplicate (n=2) at a rate of 6 mL / min, and results were calculated using a duration of 1.5 to 2.2 seconds.

[0338] The results of the syringe force experiments (n=2, 6 mL / min, average of two samples tested) are shown in Table 13 along with the density, viscosity, and UV recovery. [Table 13]

[0339] As shown in Table 13, Formulation 16, which has ethyl oleate as the liquid carrier, has the lowest syringe force of the formulations tested. Formulations 12 and 13 exhibited similar glide forces.

[0340] Figure 39, panels a-c, depict graphs of syringe force profiles for each of Formulations 12, 13, and 16. Panel a depicts the syringe force profile for Formulation 12 (benzyl benzoate as the liquid carrier). Panel b depicts the syringe force profile for Formulation 13 (Miglyol 840 as the liquid carrier). Panel c depicts the syringe force profile for Formulation 16 (ethyl oleate as the liquid carrier).

[0341] Table 14 shows the average glide force results for formulations 12, 13, and 16 compared to the same measurements for BSA and MoNi suspensions (460 mg / mL) in various other liquid carriers prepared in prefilled syringes and stored at ambient temperature away from light for 48 hours. [Table 14]

[0342] As shown in Table 14, Formulation 16, which has ethyl oleate as the liquid carrier, has the lowest syringe force of the formulations tested. Formulations 12 and 13 also exhibited lower mean glide forces than the remaining suspension formulations tested.

[0343] 6.1.18. Example 18 - BSA and MoNi Suspension in a Mixture of Triacetin and DMAc The syringe force profile of an exemplary injectable formulation containing BSA and MoNi suspension (460 mg / mL) in triacetin:DMAc (75:25) as the liquid carrier was investigated. The formulation in triacetin:DMAc (75:25) was prepared according to General Method B (described herein). Table 15 below summarizes the formulations prepared for this example. [Table 15]

[0344] Formulation 17 was loaded into prefilled syringes in duplicate (n=2) and then evaluated to establish the syringe force profile. Prefilled syringes (0.3 mL) were subjected to force measurements, and the glide force (N) was calculated over a period of 1 to 2.2 seconds.

[0345] The results of the syringe force experiments (n=2, average of two samples tested) are shown in Table 16. [Table 16]

[0346] FIG. 40 is a graph showing the syringe force profile of Formulation 17.

[0347] 7. Equivalents and Incorporation by Reference While various embodiments of the present disclosure have been particularly shown and described with reference to preferred and various alternative embodiments, it will be understood by those skilled in the art that various changes in form and detail can be made therein without departing from the spirit and scope of the present disclosure.

[0348] All publications, patents, patent applications, and other documents cited in this application, including U.S. Provisional Application Nos. 63 / 437,239 and 63 / 532,820 and International Publication Nos. 2021 / 211976 and 2023 / 230046, are incorporated herein 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. 1. An injectable pharmaceutical composition comprising: A particle, Biopharmaceutical agents, and a particle comprising a polyacrylamide-based copolymer; and a liquid carrier in which the particles are suspended.

2. 10. The injectable pharmaceutical composition of claim 1, wherein the particles have an average diameter of 100 microns or less.

3. 3. The injectable pharmaceutical composition of claim 2, wherein the particles have an average diameter of 0.01 to 100 microns.

4. 4. The injectable pharmaceutical composition of claim 3, wherein the particles have an average diameter of 0.1 to 100 microns.

5. 5. The injectable pharmaceutical composition of claim 4, wherein the particles have an average diameter of 0.2 to 20 microns.

6. 6. The injectable pharmaceutical composition of claim 5, wherein the particles have an average diameter of 0.2 to 10 microns.

7. 10. The injectable pharmaceutical composition of claim 1, further comprising one or more of a stabilizer, a preservative, a filler, a bulking agent, a sugar, a polysaccharide, or a viscosity modifier.

8. 8. The injectable pharmaceutical composition of claim 7, further comprising a stabilizer.

9. 9. The injectable pharmaceutical composition of claim 8, wherein the stabilizer is selected from surfactants, poloxamers, povidone, polyvinylpyrrolidone (PVP) polymers, polyvinyl alcohol (PVA) polymers, polysaccharides, celluloses, amphoteric compounds, sugars, salts, and combinations thereof.

10. The injectable pharmaceutical composition of any one of claims 1 to 9, wherein the biopharmaceutical agent is a polypeptide.

11. 11. The injectable pharmaceutical composition of claim 10, wherein the polypeptide is prone to aggregation in aqueous media.

12. 12. The injectable pharmaceutical composition of claim 10 or 11, wherein the polypeptide is selected from an antibody and a fragment thereof, a cytokine, a chemokine, a hormone, a vaccine antigen, a cancer antigen, an adjuvant, and combinations thereof.

13. The injectable pharmaceutical composition according to any one of claims 10 to 12, wherein the polypeptide is a protein.

14. 14. The injectable pharmaceutical composition of claim 13, wherein the protein is an antibody or a fragment thereof.

15. 15. The injectable pharmaceutical composition of claim 14, wherein the protein is a monoclonal antibody, a polyclonal antibody, an immunoglobulin G (IgG) antibody, an IgA antibody, an IgM antibody, an Fc fusion protein, or a fragment thereof.

16. The injectable pharmaceutical composition according to any one of claims 10 to 12, wherein the polypeptide is a hormone or an analog thereof.

17. 17. The injectable pharmaceutical composition of claim 16, wherein the polypeptide is insulin or an analog thereof.

18. 17. The injectable pharmaceutical composition of claim 16, wherein the polypeptide is selected from glucagon, a GLP-1 receptor agonist, amylin, and analogs thereof.

19. 19. The injectable pharmaceutical composition of any one of claims 1 to 18, wherein the composition comprises 0.1 to 20% by weight of the biopharmaceutical agent.

20. 20. The injectable pharmaceutical composition of claim 19, wherein the biopharmaceutical agent is insulin or an analog thereof.

21. 20. The injectable pharmaceutical composition of claim 19, wherein the biopharmaceutical agent is a peptide.

22. 22. The injectable pharmaceutical composition of any one of claims 19 to 21, wherein the particles comprise no more than 20% by weight of the biopharmaceutical agent.

23. 19. The injectable pharmaceutical composition of any one of claims 1 to 18, wherein the composition comprises 20% by weight or more of the biopharmaceutical agent.

24. 24. The injectable pharmaceutical composition of claim 23, wherein the composition comprises 20% to 80% by weight of the biopharmaceutical agent.

25. 25. The injectable pharmaceutical composition of claim 23 or 24, wherein the biopharmaceutical agent is an antibody or fragment thereof or an Fc fusion protein.

26. 26. The injectable pharmaceutical composition of any one of claims 23 to 25, wherein the particles comprise 75% by weight or more of the biopharmaceutical agent.

27. 27. The injectable pharmaceutical composition of any one of claims 1 to 26, wherein the composition comprises 0.01 to 20% by weight of the polyacrylamide-based copolymer.

28. 28. The injectable pharmaceutical composition of claim 27, wherein the composition comprises 0.1 to 10% by weight of the polyacrylamide-based copolymer.

29. 28. The injectable pharmaceutical composition of claim 27, wherein the composition comprises 0.5 to 5% by weight of the polyacrylamide-based copolymer.

30. 30. The injectable pharmaceutical composition of any one of claims 1 to 29, wherein the particles comprise 0.01 to 25% by weight of the polyacrylamide-based copolymer.

31. 31. The injectable pharmaceutical composition of claim 30, wherein the particles comprise 0.1 to 10% by weight of the polyacrylamide-based copolymer.

32. 31. The injectable pharmaceutical composition of claim 30, wherein the particles comprise 1 to 5% by weight of the polyacrylamide-based copolymer.

33. 33. The injectable pharmaceutical composition of any one of claims 1 to 32, wherein the composition comprises a weight to weight ratio of said polyacrylamide-based copolymer to said biopharmaceutical agent of from 1:500 to 2:

1.

34. 34. The injectable pharmaceutical composition of claim 33, wherein the composition comprises a weight to weight ratio of said polyacrylamide-based copolymer to said biopharmaceutical agent of 1:50 to 1:

1.

35. 34. The injectable pharmaceutical composition of claim 33, wherein the composition comprises a weight to weight ratio of said polyacrylamide-based copolymer to said biopharmaceutical agent of 1:25 to 1:

10.

36. The polyacrylamide copolymer is a water-soluble carrier monomer selected from N-(3-methoxypropyl)acrylamide (MPAM), 4-acryloylmorpholine (MORPH), N,N-dimethylacrylamide (DMA), N-hydroxyethylacrylamide (HEAM), acrylamide (AM), and combinations thereof; and 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-N-diethylacrylamide (DEA), N-tert-butylacrylamide (TBA), N-phenylacrylamide (PHE), and combinations thereof.

37. 34. The injectable pharmaceutical composition of claim 33, wherein the water-soluble carrier monomer is selected from MORPH, MPAM, and combinations thereof.

38. 35. The injectable pharmaceutical composition of claim 34, wherein the water-soluble carrier monomer comprises MORPH.

39. 35. The injectable pharmaceutical composition of claim 34, wherein the water-soluble carrier monomer comprises MPAM.

40. 37. The injectable pharmaceutical composition of any one of claims 33 to 36, wherein the functional dopant monomer is selected from AMP, TMA, TBA, PHE, and combinations thereof.

41. 37. The injectable pharmaceutical composition of any one of claims 33 to 36, wherein the functional dopant monomer is selected from DEA, PHE, NIP, and combinations thereof.

42. 37. The injectable pharmaceutical composition of any one of claims 33 to 36, wherein the functional dopant monomer comprises TRI.

43. 37. The injectable pharmaceutical composition of any one of claims 33 to 36, wherein the functional dopant monomer comprises PHE.

44. 37. The injectable pharmaceutical composition of any one of claims 33 to 36, wherein the functional dopant monomer comprises NIP.

45. 37. The injectable pharmaceutical composition of any one of claims 33 to 36, wherein the functional dopant monomer comprises DEA.

46. the water-soluble carrier monomer is selected from MPAM, MORPH, and combinations thereof; 34. The injectable pharmaceutical composition of claim 33, wherein the functional dopant monomer is selected from NIP, PHE, and combinations thereof.

47. the water-soluble carrier monomer is selected from MPAM, MORPH, and combinations thereof; 34. The injectable pharmaceutical composition of claim 33, wherein the functional dopant monomer is selected from AMP, TMA, TBA, PHE, and combinations thereof.

48. 34. The injectable pharmaceutical composition of claim 33, wherein the water-soluble carrier monomer is MPAM and the functional dopant monomer is PHE.

49. 34. The injectable pharmaceutical composition of claim 33, wherein the water-soluble carrier monomer is MORPH and the functional dopant monomer is PHE.

50. 34. The injectable pharmaceutical composition of claim 33, wherein the water-soluble carrier monomer is MORPH and the functional dopant monomer is NIP.

51. The polyacrylamide copolymer is 70% to 98% by weight of the water-soluble carrier monomer; 51. The injectable pharmaceutical composition of any one of claims 33 to 50, comprising 2% to 30% by weight of said functional dopant monomer.

52. The polyacrylamide copolymer is 80% to 95% by weight of the water-soluble carrier monomer; 51. The injectable pharmaceutical composition of any one of claims 33 to 50, comprising 5% to 20% by weight of said functional dopant monomer.

53. The polyacrylamide copolymer is 83% to 98% by weight of the water-soluble carrier monomer; 51. The injectable pharmaceutical composition of any one of claims 33 to 50, comprising 2% to 17% by weight of said functional dopant monomer.

54. The polyacrylamide copolymer is 70% to 85% by weight of MORPH; 34. The injectable pharmaceutical composition of claim 33, comprising 15% to 30% by weight of NIP.

55. The polyacrylamide copolymer is 74% to 80% by weight of MORPH; 20% to 26% by weight of NIP.

56. The polyacrylamide copolymer is 77 wt% MORPH; 23% by weight of NIP.

57. 57. The injectable pharmaceutical composition according to any one of claims 33 to 56, wherein the degree of polymerization of the polyacrylamide-based copolymer is 10 to 500.

58. 58. The injectable pharmaceutical composition of claim 57, wherein the degree of polymerization of the polyacrylamide-based copolymer is 20-200.

59. 59. The injectable pharmaceutical composition of claim 58, wherein the degree of polymerization of the polyacrylamide-based copolymer is 50.

60. 60. The injectable pharmaceutical composition of any one of claims 22 to 59, wherein the polyacrylamide-based copolymer has a number average molecular weight of 1,000 g / mol to 40,000 g / mol.

61. 61. The injectable pharmaceutical composition of claim 60, wherein the number average molecular weight of the polyacrylamide-based copolymer is from 2,000 g / mol to 10,000 g / mol.

62. 62. The injectable pharmaceutical composition of claim 61, wherein the number average molecular weight of the polyacrylamide-based copolymer is 4,000 g / mol to 6,000 g / mol.

63. 63. The injectable pharmaceutical composition of any one of claims 1 to 62, wherein the polyacrylamide-based copolymer is amphiphilic.

64. 64. The injectable pharmaceutical composition of any one of claims 1 to 63, wherein the liquid carrier comprises a mixture of an organic solvent and an aqueous solution.

65. 64. The injectable pharmaceutical composition of any one of claims 1 to 63, wherein the liquid carrier comprises an organic solvent, an oil, or a combination thereof.

66. 66. The injectable pharmaceutical composition of any one of claims 1 to 65, wherein the liquid carrier comprises one or more liquids selected from triacylglycerides, diacylglycerides, monoacylglycerides, acetamides, alkyl alcohols, aryl alcohols, aralkyl alcohols, fatty acids, fatty acid esters, oils, alkanes, perfluoroalkanes, propylene glycol monoesters, propylene glycol diesters, butylene glycol monoesters, butylene glycol diesters, polyethylene glycol diesters, and combinations thereof.

67. 67. The injectable pharmaceutical composition of claim 66, wherein the liquid carrier comprises an aralkyl benzoate.

68. 68. The injectable pharmaceutical composition of claim 67, wherein the aralkyl benzoate is benzyl benzoate.

69. 67. The injectable pharmaceutical composition of claim 66, wherein the liquid carrier comprises a triacylglyceride, a diacylglyceride, a monoacylglyceride, or a combination thereof.

70. 70. The injectable pharmaceutical composition of claim 69, wherein the liquid carrier comprises a triacylglyceride that is triacetin.

71. 67. The injectable pharmaceutical composition of claim 66, wherein the liquid carrier comprises N,N-dimethylacetamide (DMAc).

72. 67. The injectable pharmaceutical composition of claim 66, wherein the liquid carrier comprises a fatty acid ester.

73. 73. The injectable pharmaceutical composition of claim 72, wherein the fatty acid ester is ethyl oleate.

74. 67. The injectable pharmaceutical composition of claim 66, wherein the liquid carrier comprises a propylene glycol diester and / or a butylene glycol diester.

75. 75. The injectable pharmaceutical composition of claim 74, wherein the liquid carrier comprises Miglyol 840.

76. 67. The injectable pharmaceutical composition of claim 65 or 66, wherein the liquid carrier comprises a blend of a triacylglyceride and one or more organic solvents having a lower viscosity than the triacylglyceride.

77. 77. The injectable pharmaceutical composition of claim 76, wherein the triacylglyceride is triacetin.

78. 78. The injectable pharmaceutical composition of claim 77, wherein the liquid carrier comprises a volume-to-volume (v / v) ratio of triacetin to one or more organic solvents having a lower viscosity than triacetin of 1:1 to 9:

1.

79. 77. The injectable pharmaceutical composition of claim 76, wherein the liquid carrier comprises a 1:1 v / v ratio of triacetin to one or more organic solvents having a lower viscosity than triacetin.

80. 77. The injectable pharmaceutical composition of claim 76, wherein the liquid carrier comprises a 3:1 v / v ratio of triacetin to one or more organic solvents having a lower viscosity than triacetin.

81. 77. The injectable pharmaceutical composition of claim 76, wherein the liquid carrier comprises a 9:1 v / v ratio of triacetin to one or more organic solvents having a lower viscosity than triacetin.

82. 82. The injectable pharmaceutical composition of any one of claims 76 to 81, wherein the organic solvent having a lower viscosity than triacetin is selected from acetamide, alkyl benzoate, aryl benzoate, aralkyl benzoate, aryl alcohol, aralkyl alcohol, or any combination thereof.

83. 83. The injectable pharmaceutical composition of claim 82, wherein the organic solvent having a lower viscosity than triacetin is acetamide.

84. 84. The injectable pharmaceutical composition of claim 83, wherein the acetamide is DMAc.

85. 83. The injectable pharmaceutical composition of claim 82, wherein the organic solvent having a lower viscosity than triacetin is an aralkyl alcohol.

86. 86. The injectable pharmaceutical composition of claim 85, wherein the aralkyl alcohol is benzyl alcohol.

87. 83. The injectable pharmaceutical composition of claim 82, wherein the organic solvent having a lower viscosity than triacetin is an aralkyl benzoate.

88. 88. The injectable pharmaceutical composition of claim 87, wherein the aralkyl benzoate is benzyl benzoate.

89. 83. The injectable pharmaceutical composition of claim 82, wherein the organic solvent having a lower viscosity than triacetin comprises a combination of acetamide and an aralkyl alcohol.

90. 90. The injectable pharmaceutical composition of claim 89, wherein the acetamide is DMAc and the aralkyl alcohol is benzyl alcohol.

91. 91. The injectable pharmaceutical composition of claim 90, wherein the low viscosity organic solvent comprises a v / v ratio of DMAc to benzyl alcohol of 3:1 to 2:

1.

92. 67. The injectable pharmaceutical composition of claim 65 or 66, wherein the organic solvent is an alkyl benzoate, an aryl benzoate, an aralkyl benzoate, or any combination thereof.

93. 93. The injectable pharmaceutical composition of claim 92, wherein the organic solvent is benzyl benzoate.

94. 66. The injectable pharmaceutical composition of claim 64 or 65, wherein the organic solvent is a polar aprotic solvent.

95. 95. The injectable pharmaceutical composition of claim 94, wherein the polar aprotic solvent is dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), or a mixture thereof.

96. 66. The injectable pharmaceutical composition of claim 65, wherein the liquid carrier comprises an oil (e.g., a vegetable oil or an animal oil).

97. 97. The injectable pharmaceutical composition of claim 96, wherein the oil is coconut oil, cottonseed oil, fish oil, grapeseed oil, hazelnut oil, hydrogenated vegetable oil, lime oil, olive oil, palm seed oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, walnut oil, or any combination thereof.

98. 98. The injectable pharmaceutical composition of claim 97, wherein the oil is sesame oil.

99. 99. An injectable pharmaceutical composition according to any one of claims 1 to 98, wherein the composition comprises a weight to weight ratio of the liquid carrier to the particles of from 1:5 to 4:

1.

100. 100. The injectable pharmaceutical composition of claim 99, wherein the composition comprises a weight to weight ratio of the liquid carrier to the particles of from 1:1 to 2:

1.

101. 1. An injectable pharmaceutical composition comprising: A particle, the particle comprising: Biopharmaceutical agents, and a polyacrylamide-based copolymer, wherein 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), acrylamide (AM), and combinations thereof; and particles comprising 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-N-diethylacrylamide (DEA), N-tert-butylacrylamide (TBA), N-phenylacrylamide (PHE), and combinations thereof; an injectable pharmaceutical composition comprising: a liquid carrier in which the particles are suspended, the liquid carrier being selected from a triacylglyceride, a diacylglyceride, a monoacylglyceride, an acetamide, an alkyl alcohol, an aryl alcohol, an aralkyl alcohol, a fatty acid, a fatty acid ester, an oil, an alkane, a perfluoroalkane, a propylene glycol monoester, a propylene glycol diester, a butylene glycol monoester, a butylene glycol diester, a polyethylene glycol diester, and combinations thereof.

102. 102. The injectable pharmaceutical composition of claim 101, wherein the water-soluble carrier monomer is selected from MORPH, MPAM, and combinations thereof.

103. 103. The injectable pharmaceutical composition of claim 101 or 102, wherein the functional dopant monomer is selected from AMP, TMA, TBA, PHE, and combinations thereof.

104. 103. The injectable pharmaceutical composition of claim 101 or 102, wherein the functional dopant monomer is selected from DEA, PHE, NIP, and combinations thereof.

105. the water-soluble carrier monomer is selected from MPAM, MORPH, and combinations thereof; 105. The injectable pharmaceutical composition of claim 104, wherein the functional dopant monomer is selected from NIP, PHE, and combinations thereof.

106. 105. The injectable pharmaceutical composition of claim 104, wherein the water-soluble carrier monomer is MORPH and the functional dopant monomer is NIP.

107. 107. The injectable pharmaceutical composition of any one of claims 101 to 106, wherein the liquid carrier comprises an aralkyl benzoate.

108. 108. The injectable pharmaceutical composition of claim 107, wherein the aralkyl benzoate is benzyl benzoate.

109. 107. The injectable pharmaceutical composition of any one of claims 101 to 106, wherein the liquid carrier comprises a triacylglyceride.

110. 110. The injectable pharmaceutical composition of claim 109, wherein the triacylglyceride is triacetin.

111. 107. The injectable pharmaceutical composition of any one of claims 101 to 106, wherein the liquid carrier comprises N,N-dimethylacetamide (DMAc).

112. 107. The injectable pharmaceutical composition of any one of claims 101 to 106, wherein the liquid carrier comprises a fatty acid ester.

113. 113. The injectable pharmaceutical composition of claim 112, wherein the fatty acid ester comprises ethyl oleate.

114. 107. The injectable pharmaceutical composition of any one of claims 101 to 106, wherein the liquid carrier comprises a propylene glycol diester and / or a butylene glycol diester.

115. the propylene glycol diester is Miglyol 840; 115. The injectable pharmaceutical composition of claim 114, wherein the butylene glycol diester is Miglyol 8810.

116. A syringe loaded with the composition of any one of claims 1 to 115.

117. 117. The syringe of claim 116, configured to eject the composition at a flow rate of 0.1 mL / min or greater in response to a force applied to the syringe of 70 N or less.

118. 117. The syringe of claim 116, configured to eject the composition at a flow rate of 0.1 mL / min or greater in response to a force applied to the syringe of 50 N or less.

119. A syringe according to any one of claims 116 to 118, wherein the syringe is equipped with a needle having a size of 18 to 32 gauge (G).

120. 120. The syringe of claim 119, wherein the needle is a 22G needle.

121. 120. The syringe of claim 119, wherein the needle is a 24G needle.

122. 120. The syringe of claim 119, wherein the needle is a 25G needle.

123. 120. The syringe of claim 119, wherein the needle is a 26G needle.

124. 120. The syringe of claim 119, wherein the needle is a 27G needle.

125. 120. The syringe of claim 119, wherein the needle is a 30G needle.

126. 120. The syringe of claim 119, wherein the needle is a 32G needle.

127. A syringe according to any one of claims 119 to 126, wherein the needle is an ultra-thin-walled needle.

128. A method for administering a biopharmaceutical to a subject by injection, comprising:

116. A method for administering a therapeutically effective dose of said biopharmaceutical to a subject in need thereof, comprising injecting the composition of any one of claims 1 to 115.

129. 129. The method of claim 128, wherein the injecting is performed using a loaded syringe according to any one of claims 116 to 127.

130. 1. A method for preparing an injectable pharmaceutical composition, comprising: d) a biopharmaceutical agent; providing a mixture comprising: a polyacrylamide-based copolymer in water; e) removing the water from the mixture to obtain particles comprising the biopharmaceutical agent and the polyacrylamide-based copolymer; f) contacting a liquid carrier with said particles to form a suspension of said particles in said liquid carrier.

131. 131. The method of claim 130, wherein in step b), the water is removed via spray drying.

132. 131. The method of claim 130, wherein in step b), the water is removed via electrospray drying.

133. 131. The method of claim 130, wherein in step b), the water is removed via freeze-drying.

134. 131. The method of claim 130, wherein the method further comprises a milling step after the freeze-drying but before step c).

135. 135. A method according to any one of claims 130 to 134, wherein the particles obtained in step b) have an average diameter of 100 microns or less.

136. 136. The method of any one of claims 130 to 135, wherein step c) comprises sonicating the particles in the liquid carrier.

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