Compositions and methods for delivery of therapeutic peptides

Abstract

Disclosed herein are compositions and methods for delivering therapeutic peptides. In some embodiments, for example, a composition for treating a disease or condition comprises a dynamic hydrogel and an acylated peptide encapsulated by the dynamic hydrogel. The dynamic hydrogel may include a polymer and a plurality of nanoparticles. The polymer may be non-covalently crosslinked with the plurality of nanoparticles.

Description

[0001] Cross-reference to related applications

[0002] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 528,525, filed July 24, 2023, and U.S. Provisional Application No. 63 / 569,651, filed March 25, 2024, the contents of each of which are incorporated herein by reference in their entirety.

[0003] By referencing the merged sequence list

[0004] This application contains an electronic sequence list in XML file format named “APL_010WO_SL.xml”, created on July 9, 2024, and 2,650 bytes in size, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0005] This technology generally relates to drug delivery, and more particularly to compositions and methods for delivering therapeutic peptides.

[0006] background

[0007] Diabetes, or prediabetes, affects approximately 500 million people worldwide, including an estimated 130 million individuals in the United States. In the US alone, the total annual cost directly related to diabetes and prediabetes is approximately $400 billion, making it the tenth most expensive disease in the country. Type 2 diabetes (T2D), accounting for 90-95% of all diabetes cases, is a metabolic disorder characterized by insulin resistance, pancreatic beta-cell dysfunction, and impaired regulation of glucose production in the liver, ultimately leading to beta-cell failure. Poorly managed T2D patients are at risk of serious microvascular and macrovascular complications, including cardiovascular disease, kidney disease, retinopathy, neuropathy, and stroke.

[0008] Current insulin therapy for type 2 diabetes (T2D) is very cumbersome, leading to poor patient adherence and sometimes causing dangerous hypoglycemic events. In contrast, treatment strategies based on incretin mimics (which mimic the natural incretin hormone secreted after carbohydrate intake) eliminate the risk of hypoglycemia. These treatments reduce endogenous glucose production and drive the expansion of insulin-secreting β-cells to restore the patient's natural ability to regulate blood glucose. However, conventional treatment with incretin mimics involves daily or weekly injections, which is a significant patient burden and leads to poor adherence. Brief description of the attached diagram

[0010] Many aspects of this disclosure can be better understood with reference to the following figures. The components in the figures are not necessarily drawn to scale. Rather, the focus is on clearly illustrating the principles of this disclosure.

[0011] Figure 1Aand Figure 1B A PNP hydrogel for prolonging the delivery of therapeutic peptides is shown. Figure 1A This is a schematic diagram of a polymer nanoparticle (PNP) hydrogel prepared by mixing hydrophobically modified hydroxypropyl methylcellulose (HPMC) with poly(ethylene glycol)-block-poly(lactic acid) (PEG-PLA) nanoparticles, which allows for easy encapsulation of therapeutic peptides. Figure 1B This is a schematic diagram of the formation of a local reservoir in the subcutaneous space after subcutaneous injection of PNP hydrogel, which can provide an adjustable platform for the sustained release of therapeutic peptides.

[0012] Figure 2A This graph indicates that once-weekly dosing frequency did not significantly improve patient compliance compared to once-daily dosing frequency.

[0013] Figure 2B This is a graph showing clinical data on the release curves of conventional incretin mimicry therapy. In the lower graph, the dashed line represents patients receiving repeated weekly injections for four months to achieve therapeutic concentrations of the incretin mimicry. The standard treatment strategy involves daily or weekly subcutaneous injections with significantly escalating time to achieve therapeutic concentrations. In contrast, the solid line in the upper graph represents the target delivery curve for a single PNP hydrogel reservoir injection providing continuous release of the incretin mimicry for 120 days. The dashed lines in both graphs indicate the treatment threshold.

[0014] Figure 3A This is a graph showing the size exclusion chromatography (SEC) trace of the PEG-PLA polymer. Gel permeation chromatography characterization of the PEG-PLA polymer shows a single peak corresponding to the PEG-PLA block copolymer (Mn = 22 kDa). ).

[0015] Figure 3B This is a graph showing the dynamic light scattering (DLS) data of PEG-PLA nanoparticles after nanoprecipitation (DH = 33.2, PDI = 0.038).

[0016] Figure 4 A series of photographs illustrating the preparation of the PNP hydrogel are shown. The PNP hydrogel was prepared by mixing a solution of hydrophobically modified HPMC (“polymer solution”, syringe on the right) with a solution of PEG-PLA nanoparticles and a therapeutic peptide (e.g., semaglutide) (“NP solution”, syringe on the left) using a Luerlock mixer. After mixing, a homogeneous, bubble-free, solid-like PNP hydrogel was formed. Due to their dynamic cross-linking, the PNP hydrogel can be injected with clinically relevant high-specification needles and rapidly self-heals after injection.

[0017] Figure 5A and Figure 5B This is a graph showing the rheological characterization of the PNP-1-10 hydrogel formulation with varying semaglutide concentrations: frequency-dependent oscillatory shear scan ( Figure 5A ) and stress-dependent oscillatory shear scan ( Figure 5B ).

[0018] Figure 6A and Figure 6B This is a graph showing the rheological characterization of the PNP-2-10 hydrogel formulation with varying semaglutide concentrations: frequency-dependent oscillatory shear scan ( Figure 6A ) and stress-dependent oscillatory shear scan ( Figure 6B ).

[0019] Figures 7A-7C This is a graph showing the rheological characterization of the PNP-1-10 hydrogel formulation with varying tirzepatide concentrations: frequency-dependent oscillatory shear scan. Figure 7A and Figure 7C ) and stress-dependent oscillatory shear scan ( Figure 7B ).

[0020] Figure 8A This is a schematic diagram of the in vitro release assay of smegglutinin from a PNP hydrogel immersed in saline solution over a two-week period. The in vitro release assay was designed to minimize hydrogel erosion by minimizing the surface area to volume ratio of the hydrogel.

[0021] Figure 8B This is a graph showing the in vitro release curves of the cumulative release % of semaglutide from the PNP-2-10 formulation over a two-week period at low, medium, and high semaglutide loadings.

[0022] Figure 8C This is a graph showing the in vitro release curves of the cumulative release percentage of semaglutide from the PNP-1-10 formulation over a two-week period at low, medium, and high semaglutide loadings.

[0023] Figure 8D This is a graph showing the in vitro release curves of the cumulative release % of semaglutide from various PNP-1-10 hydrogel formulations over a two-week period, which shows the effect of the addition of Tween 20 on semaglutide release in the presence of propylene glycol and saline.

[0024] Figure 8EThe graph shows the in vitro release curves of smegglutinin from various PNP-1-10 hydrogel formulations over a two-week period, demonstrating that the addition of α-cyclodextrin has a negligible effect on smegglutinin release in the presence of propylene glycol and saline.

[0025] Figure 8F This is a graph showing the in vitro release curves of the cumulative release % of semaglutide from various PNP-1-10 hydrogel formulations over a two-week period, which shows the effect of adding Tween 20 on semaglutide release in the presence of propylene glycol and in the absence of saline.

[0026] Figure 8G This is a graph showing the in vitro release curves of smegglutinin from various PNP-1-10 hydrogel formulations, illustrating the cumulative release percentage, and demonstrating the role of bovine serum albumin (BSA) in the release buffer and hydrogel.

[0027] Figure 9 This is a graph showing the in vitro release curves of liraglutide from 1.8 mg / mL PNP-1-10 and PNP-2-10 hydrogel formulations over a two-week period, representing the cumulative release percentage.

[0028] Figure 10 This is a graph showing the in vitro release curves of the cumulative release % of liraglutide and insulin glargine (Lantus) from the PNP-1-10 hydrogel formulation over a two-week period.

[0029] Figure 11 This is a schematic diagram illustrating the treatment plan and timing for blood glucose measurement and serum collection used in the analysis. Diabetic rats received a single subcutaneous injection of PNP hydrogel loaded with semaglutide (PNP-1-10 with 1.8 mg / mL semaglutide and 0.05 wt% Tween 20) or PNP hydrogel loaded with telpolide (PNP-1-10 with 4.5 mg / mL telpolide and 0.05 wt% Tween 20), or daily subcutaneous bolus injections of PBS, 20 µg semaglutide, or 50 µg telpolide.

[0030] Figure 12A and Figure 12B This describes the pre-treatment ( Figure 12A ) and 6 weeks after treatment ( Figure 12BA graph showing the results of the oral glucose tolerance test. An oral glucose tolerance test (OGTT) was performed to classify diabetic rats into treatment groups. Rats were fasted before administration of a glucose load via oral gavage. Baseline (fasting) blood glucose was measured before glucose administration, and then at regular intervals. Blood glucose was measured at -5 min, 0 min, 5 min, 15 min, 30 min, 45 min, 60 min, and 120 min. Rats with similar glucose tolerance were paired using the area under the curve (AUC) and then randomly assigned to treatment groups.

[0031] Figure 13 This is a graph showing the percentage change in blood glucose (BG) levels in rats treated with PNP hydrogel relative to those treated with bolus injection. A single administration of semaglutide PNP or telposide PNP hydrogel reduced BG in type 2 diabetes-like male rats over a 6-week period compared to daily PBS bolus injections. This graph shows the change in BG over 6 weeks after each treatment regimen (n = 6).

[0032] Figure 14 This is a graph showing the percentage change in body weight in rats treated with PNP hydrogel relative to those treated with bolus injection. A single administration of semaglutide PNP or telpolide PNP hydrogel reduced the total body weight gain in type 2 diabetes-like male rats over the 6-week treatment period compared to daily PBS bolus injections. This graph shows the change in body weight over 6 weeks in each treatment group (n = 6).

[0033] Figure 15 This is a graph showing the pharmacokinetics of 20 µg daily bolus injection of semaglutide relative to PNP hydrogel (PNP-1-10 with 1.8 mg / mL semaglutide loading and 0.05 wt% Tween 20) in male diabetic rats (n = 6) over a period of 6 weeks following treatment.

[0034] Figure 16 This is a graph showing the pharmacokinetics of 20 µg daily bolus injection of semaglutide relative to PNP hydrogel (PNP-1-10 with 1.8 mg / mL semaglutide loading and 0.05 wt% Tween 20) over 24 hours in male diabetic rats (n = 6), superimposed on the pharmacokinetics of 20 µg intravenous (IV) and subcutaneous (SC) bolus injections.

[0035] Figure 17The diagram illustrates the use of blood chemistry to assess treatment biocompatibility, observe negative effects on the liver or kidneys, and evaluate the treatment's efficacy against HbA1c. Blood was collected before and after treatment (6 weeks later). Hepatotoxicity was assessed by measuring alanine aminotransferase (ALT), aspartate aminotransferase (AST), and bilirubin. Nephrotoxicity was assessed by examining creatinine and blood urea nitrogen (BUN) levels. For both treatment and control groups, ALT, AST, creatine, and BUN values ​​were within the range of healthy rats (defined as mean ± 2 standard deviations).

[0036] Detailed Explanation

[0037] Therapeutic peptides that mimic the activity of naturally occurring peptides can be used to treat a variety of diseases and conditions. For example, glucagon-like peptide-1 (GLP-1) is an incretin hormone and neurotransmitter secreted by intestinal L-cells in response to nutrients, stimulating insulin in a glucose-dependent manner and blocking glucagon secretion. GLP-1 itself degrades rapidly, but because its beneficial effects extend beyond glucose control, long-acting GLP-1 receptor agonists (GLP-1 RAs) have been developed for the treatment of type 2 diabetes. However, patient adherence to anti-glycemic medications is surprisingly low, described as falling between 29% and 54% for GLP-1 RAs, resulting in suboptimal type 2 diabetes management, which carries an increased risk of stroke, heart and kidney disease, amputation, and blindness. For drugs with short half-lives, poor adherence to prescribed treatment regimens lowers plasma concentrations to inappropriate levels and often requires multiple doses to restore therapeutic plasma concentrations. The complexity and / or frequency of treatment administration is a barrier to adherence. Conventionally, GLP-1 RA treatment agents are injected daily or weekly or taken orally daily, leaving room for technological innovation that reduces the frequency of administration, which can alleviate the burden on patients and increase patient compliance.

[0038] To address these and other challenges, this technology provides compositions and methods for delivering therapeutic peptides, such as incretin mimics (e.g., GLP-1 RA) and / or acylated peptides. In some embodiments, for example, this disclosure provides compositions for treating diseases or conditions (e.g., diabetes and / or obesity), wherein the composition comprises a dynamic hydrogel consisting of a polymer (e.g., a hydrophobically modified cellulose derivative) and multiple nanoparticles (e.g., amphiphilic polymeric nanoparticles). The polymer may be non-covalently crosslinked with the multiple nanoparticles, thereby imparting shear-thinning, self-healing, and / or viscoelastic properties to the dynamic hydrogel. The composition may also comprise acylated peptides, such as incretin mimics, encapsulated by the dynamic hydrogel. In some embodiments, the acylated peptide is encapsulated via a hydrophobic interaction between the fatty acid side chains of the acylated peptide and the hydrophobic surface of the nanoparticles. The acylated peptide can be gradually released from the dynamic hydrogel via erosion of the dynamic hydrogel in vivo. Therefore, after administration of the composition to a subject, the composition can provide sustained and controlled release of the acylated peptide at a rate that effectively treats the disease or condition for the desired therapeutic period. For example, the compositions described herein can be designed to provide continuous delivery of an incretin mimic for more than four months from a single administration, to match the cycle in which patients with type 2 diabetes typically visit their doctors.

[0039] Compared to conventional therapeutic products and methods, the implementation of this technology offers numerous advantages. For example, conventional hydrogel-based reservoir technologies typically exhibit several significant drawbacks, including complex manufacturing, poor formulation stability, challenging administration, burst release that may lead to poor treatment tolerance, and slow release that is insufficient for achieving adequate long-acting therapy. In contrast to conventional covalently cross-linked hydrogels, the dynamic hydrogels of this technology are formed through strong and dynamic physical interactions. Therefore, these materials can overcome the shortcomings of other hydrogel-based reservoir technologies by exhibiting the following characteristics: (i) mild formulation requirements, which facilitate easy formulation with therapeutic peptides (e.g., incretin mimics) and maintain drug stability during manufacturing and storage; (ii) shear-thinning properties, which allow for direct injection via standard syringes and needles, thereby improving patient convenience; (iii) rapid self-healing and reservoir formation of the hydrogel structure to avoid burst release of the drug cargo, thus providing excellent tolerability by maintaining a consistent, slow release and avoiding undesirable side effects (e.g., gastrointestinal discomfort); (iv) sufficiently high yield stress to form a robust reservoir that persists under normal stress in the subcutaneous space after administration; (v) extended delivery of the therapeutic cargo, allowing for continuous delivery within the clinically desired timeframe; (vi) biodegradability; and / or (vii) non-immunogenicity and non-promoting of immune responses to the encapsulated cargo.

[0040] Embodiments of this disclosure will be described more fully below with reference to the accompanying drawings, in which exemplary embodiments are shown. However, the embodiments of the claims may be implemented in many different forms and should not be construed as limited to the embodiments set forth herein. The examples set forth herein are non-limiting examples and are merely examples among other possible examples.

[0041] The headings provided herein are for convenience only and do not constitute an explanation of the scope or meaning of the claimed technology. Embodiments under any one heading may be used in conjunction with embodiments under any other heading.

[0042] I. Dynamic hydrogel

[0043] This technology utilizes dynamic hydrogels, which can serve as a universal platform for the controlled release of therapeutic goods (e.g., the therapeutic peptides described in Part II below). In some embodiments, the dynamic hydrogel exhibits dynamic behavior, such as shear-thinning behavior, self-healing behavior, and / or highly tunable viscoelastic mechanical properties. The shear-thinning, self-healing, and / or viscoelastic properties of the dynamic hydrogel can arise from non-covalent supramolecular interactions between the hydrogel components (e.g., polymers and nanoparticles, further described below). Non-covalent interactions can include physical crosslinking, which can encompass various types of crosslinking induced by weak physical interactions (e.g., hydrogen bonding, hydrophobic interactions, ionic interactions, van der Waals interactions, host-guest interactions, crystal formation, physical entanglement, or combinations thereof). Non-covalent interactions can allow the formation of dynamically reversible crosslinks between the hydrogel components, which can, for example, spontaneously and / or in response to applied stress, dissociate and recombine.

[0044] The dynamic hydrogels described herein offer numerous advantages for therapeutic applications. For example, they can exhibit high drug loading capacity, mild conditions for encapsulating biological cargoes, sustained delivery of cargoes, and / or mechanical tunability. However, unlike conventional covalently cross-linked hydrogels, these dynamic hydrogels can be readily applied via techniques such as direct injection, catheter delivery, spreading, or spraying due to their shear-thinning and / or self-healing properties. Furthermore, they can exhibit unique dynamic network rearrangements, providing highly tunable release characteristics for therapeutic cargoes. The dynamic hydrogels presented herein can also be synthesized in a readily scalable, cost-effective manner.

[0045] A. Polymer nanoparticle hydrogel

[0046] In some embodiments, the dynamic hydrogels described herein are polymer nanoparticle (PNP) hydrogels. PNP hydrogels are supramolecular hydrogels formed by non-covalent interactions between polymers and nanoparticles. PNP hydrogels can rapidly self-assemble when a polymer solution is mixed with a nanoparticle solution. Self-assembly of PNP hydrogel networks can occur when polymers are linked together by fragments of polymer chains adsorbed onto the surface of nanoparticles through multivalent transient interactions. PNP hydrogel formation can be an entropy-driven process, where solvent molecules (e.g., water) solvating the polymer chains and nanoparticle surfaces are released into the bulk solution as the polymer chains bind to the nanoparticle surfaces, resulting in a large translational entropy gain. The interactions between the polymer and nanoparticle surfaces can be transient and reversible, allowing the PNP hydrogel to flow under applied shear stress and subsequently self-heal rapidly upon stress relaxation.

[0047] The PNP hydrogels described herein can consist of any suitable combination of polymers and nanoparticles capable of non-covalent interactions to form crosslinked structures with desired dynamic behavior. In some embodiments, the nanoparticles and polymers are selected to have sufficiently strong affinity to produce effective crosslinking. That is, the free energy gain (c) resulting from the adsorption of polymer chains onto the nanoparticle surface can be greater than or equal to the thermal energy (kJ / kJ). B T) or with thermal energy (k) B The modulus (G) of the PNP hydrogel is comparable to that of the polymer chain. Furthermore, the average number of interactions between each polymer chain and particle can be greater than 2 to achieve percolation of the hydrogel network. Additionally, to facilitate polymer bridging of multiple nanoparticles (as opposed to polymers entangled with a single particle), the nanoparticle diameter can be comparable to or smaller than the persistence length of the polymer chain. When some or all of these criteria are met, the nanoparticles can act as crosslinking agents between polymer chains, which can bridge many different particles, thereby enabling the formation of a hydrogel. In some embodiments, the modulus (G) of the PNP hydrogel is related to the number (n) of dynamic hydrogel interactions per unit volume and the energy (αk) associated with each interaction, according to the following relationship. B T) Related: G ≈ nαk B T.

[0048] In some embodiments, the nanoparticle surface is hydrophobic, such that the adsorption of the polymer chain to the nanoparticle surface is at least partially influenced by the general level of hydrophobicity along the polymer chain (e.g., the size and / or number of hydrophobic groups attached to the polymer chain). For example, the PNP hydrogels described herein can utilize polymer-nanoparticle interactions between hydrophobically modified cellulose derivatives and nanoparticles (e.g., dodecyl-modified hydroxypropyl methylcellulose (HPMC-C12) and biodegradable polymer nanoparticles composed of poly(ethylene glycol)-block-poly(lactic acid) (PEG-PLA). Additional examples of nanoparticles and polymers in PNP hydrogels suitable for use herein are provided in Sections IA1 and IA2 below, respectively.

[0049] The PNP hydrogels described herein can be distinguished from conventional drug delivery systems that include nanoparticles embedded in covalently cross-linked hydrogels. Such conventional systems typically consist of a gel-forming polymer that is covalently cross-linked with each other to form a gel network, while the nanoparticles serve as an optional additive that does not play a role in gel formation and can therefore be freely replaced by other additives or omitted entirely. In contrast, the PNP hydrogels of this invention can be formed specifically through the interaction between the nanoparticles and the polymer. In some embodiments, the polymers and nanoparticles used in the PNP hydrogels of this invention do not form gels independently, or are not used at concentrations where either the polymer or the nanoparticles alone form a gel, such that gel formation occurs only when the polymer and nanoparticles are combined.

[0050] In some embodiments, the PNP hydrogels of this invention comprise one or more polymers combined with one or more nanoparticles, such that the loss modulus of a solution of the one or more polymers and the loss modulus of a solution of the one or more particles are each greater than their respective storage modulus at angular frequencies ranging from 0.1 rad / s to 100 rad / s (e.g., 10 rad / s), as measured by an oscillating shear rheometer in the linear viscoelastic region. The storage modulus of the PNP hydrogel produced by combining one or more polymers with one or more particles can be greater than the loss modulus of the PNP hydrogel at angular frequencies ranging from 0.1 rad / s to 100 rad / s (e.g., 10 rad / s), as measured by an oscillating shear rheometer in the linear viscoelastic region. In some embodiments, the PNP hydrogel at 0.1 s... -1 up to 100 s -1 (For example, 10s) -1 The dynamic shear viscosity at the shear rate of 0.1 s⁻¹ is greater than that at 0.1 s⁻¹. -1 up to 100 s -1The dynamic shear viscosity is the sum of the dynamic shear viscosity of a solution of one or more polymers and the dynamic shear viscosity of a solution of one or more nanoparticles at a given shear rate. For example, the dynamic shear viscosity of a PNP hydrogel can be 2 to 100,000, 2 to 1,000, 2 to 100, or 2 to 10 times greater than the sum of the dynamic shear viscosities of the polymer solution and the nanoparticle solution.

[0051] The PNP hydrogels described herein can contain any concentration of polymers and nanoparticles suitable for providing desired hydrogel properties. For example, higher polymer concentrations can produce PNP hydrogels with higher stiffness and / or slower degradation rates. Higher nanoparticle concentrations can produce PNP hydrogels with higher viscosity, stiffness, and yield stress and / or slower degradation rates. Hydrogel properties can depend not only on the total amount of solids in the hydrogel but also on the stoichiometry of polymer and nanoparticle content. For example, increasing the nanoparticle concentration at a constant polymer concentration can produce hydrogels with a more solid-like rheological response (e.g., lower tan δ), increased strain-to-yield, and increased yield stress. Increasing the polymer concentration at a constant nanoparticle concentration can produce hydrogels with a more liquid-like rheological response (e.g., higher tan δ and greater frequency dependence of storage modulus) and decreased strain-to-yield.

[0052] In some embodiments, the PNP hydrogel described herein comprises at least 0.25 wt%, 0.5 wt%, 0.75 wt%, 1 wt%, 1.5 wt%, 2 wt%, 2.5 wt%, 3 wt%, 3.5 wt%, 4 wt%, 4.5 wt%, or 5 wt% of polymer; and / or at least 0.5 wt%, 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 12 wt%, or 15 wt% of nanoparticles. Optionally or in combination, the concentration of the polymer within the PNP hydrogel can be 0.5 wt% to 5 wt%, 0.5 wt% to 4 wt%, 0.5 wt% to 3 wt%, 0.5 wt% to 2 wt%, 0.5 wt% to 1 wt%, 1 wt% to 5 wt%, 1 wt% to 4 wt%, 1 wt% to 3 wt%, 1 wt% to 2 wt%, 2 wt% to 5 wt%, 2 wt% to 4 wt%, 2 wt% to 3 wt%, 3 wt% to 5 wt%, 3 wt% to 4 wt%, or 4 wt% to 5 wt%; and / or the concentration of the nanoparticles within the PNP hydrogel can be 1 wt% to 12 wt%, 1 wt% to 10 wt%, 1 wt% to 8 wt%, 1 wt% to 5 wt%, 1 wt% to 3 wt%, 3 wt% to 12 wt%, 3 wt% to 10 wt%, 3 wt% to 8 wt%. wt%, 3 wt% to 5 wt%, 5 wt% to 12 wt%, 5 wt% to 10 wt%, 5 wt% to 8 wt%, 8 wt% to 12 wt%, 8 wt% to 10 wt%, or 10 wt% to 12 wt%. The term “XY hydrogel” or “X:Y hydrogel” is used herein to refer to a hydrogel having X wt% polymer and Y wt% nanoparticles.

[0053] In some embodiments, the PNP hydrogels described herein are prepared by simply mixing a polymer, nanoparticles, therapeutic cargo, and any optional additives. For example, the PNP hydrogel can be prepared by forming a polymer solution (e.g., by dissolving the polymer in an aqueous solvent such as water or a buffer solution such as phosphate-buffered saline (PBS),) a nanoparticle solution (e.g., by suspending the nanoparticles in an aqueous solvent), and a solution containing therapeutic cargo (e.g., by dissolving or suspending the therapeutic cargo in an aqueous solvent). The solutions can then be combined (optionally with external stirring) to form a PNP hydrogel containing the therapeutic cargo.

[0054] 1. Nanoparticles

[0055] The PNP hydrogel described in this article can contain multiple nanoparticles. The nanoparticles can be of any suitable shape, such as spheres, cubes, rods, tubes, plates, fibers, etc. The average particle size (e.g., diameter) of the nanoparticles can be 1 nm to 1000 nm, 1 nm to 500 nm, 1 nm to 250 nm, 1 nm to 150 nm, 1 nm to 100 nm, 1 nm to 50 nm, 1 nm to 25 nm, 1 nm to 10 nm, 10 nm to 1000 nm, 10 nm to 500 nm, 10 nm to 250 nm, 10 nm to 150 nm, 10 nm to 100 nm, 10 nm to 50 nm, 10 nm to 25 nm, 25 nm to 1000 nm, 25 nm to 500 nm, 25 nm to 250 nm, 25 nm to 150 nm, 25 nm to 100 nm, 25 nm to 50 nm, 50 nm to 1000 nm, 50 nm to 500 nm, 50 nm to 250 nm, 50 nm to 150 nm, 50 nm to 100 nm, 100 nm to 1000 nm. The nanoparticles have an average particle size of 500 nm to 500 nm, 100 nm to 250 nm, 100 nm to 150 nm, 150 nm to 1000 nm, 150 nm to 500 nm, 150 nm to 250 nm, 250 nm to 1000 nm, 250 nm to 500 nm, or 500 nm to 1000 nm. In some embodiments, the average particle size of the nanoparticles is less than or equal to 500 nm, 250 nm, 150 nm, 140 nm, 130 nm, 120 nm, 110 nm, 100 nm, 90 nm, 80 nm, 70 nm, 60 nm, 50 nm, 40 nm, 30 nm, 20 nm, or 10 nm. As described herein, to promote hydrogel formation, the average particle size of the nanoparticles can be similar to or smaller than the durability length of the polymer in the PNP hydrogel, for example, less than or equal to 125%, 110%, 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, or 10% of the polymer durability length. As used herein, “average particle size” can refer to the statistical average particle size (e.g., diameter) of the particles in the PNP hydrogel composition. The diameter of substantially spherical particles can refer to the physical or hydrodynamic diameter. The diameter of non-spherical particles can refer to the hydrodynamic diameter or the maximum linear distance between two points on the particle surface. Average particle size can be measured using methods known in the art, such as dynamic light scattering.

[0056] Nanoparticles can be made from a single material or from a combination of multiple different materials (e.g., two, three, four, five, or more different materials). The materials can be biodegradable and / or biocompatible. For example, in some embodiments, the nanoparticles are partially or entirely made from one or more biodegradable and / or biocompatible polymers. Typically, biodegradable polymers can be degraded by enzymatic hydrolysis, in vivo exposure to water, surface erosion, and / or bulk erosion. Biodegradable polymers can include synthetic polymers, naturally occurring polymers, or combinations thereof. Examples of synthesized biodegradable polymers include polyhydroxy acids (e.g., poly(lactic acid), poly(glycolic acid)), polyanhydrides, poly(orthoesters), polyesters, polyurethanes, poly(butyric acid), poly(valeric acid), poly(caprolactone), poly(hydroxybutyrate), poly(lactide-co-glycolic acid), poly(lactide-co-caprolactone), poly(ethylene-co-maleic anhydride), poly(ethylene maleic anhydride-co-L-dopamine), poly(ethylene maleic anhydride-co-phenylalanine), poly(ethylene maleic anhydride-co-tyrosine), poly(butadiene-co-maleic anhydride), poly(butadiene maleic anhydride-co-L-dopamine) (pBMAD), poly(butadiene maleic anhydride-co-phenylalanine), poly(butadiene maleic anhydride-co-tyrosine), and combinations thereof (e.g., mixtures, copolymers). Examples of naturally occurring biodegradable polymers include polysaccharides (e.g., cellulose, alginate, collagen, chitosan, hyaluronic acid, starch, agarose, agar, xanthan gum), proteins (e.g., collagen, fibrin, albumin, zein, gelatin) and their derivatives (e.g., cellulose derivatives, such as cellulose nanocrystals, cellulose nanofibers) and combinations thereof.

[0057] Optionally or in combination, nanoparticles may be made partially or entirely of one or more non-biodegradable polymers. Examples of non-biodegradable polymers include polystyrene, polyalkylene glycols, poly(meth)acrylates, poly(meth)acrylamide, polyolefins (e.g., polyethylene, polyvinyls, poly(vinyl acetate), poly(ethylene terephthalate)) and combinations thereof.

[0058] The polymers used to form the nanoparticles described herein can have any suitable molecular weight, such as molecular weight (e.g., number-average molecular weight (M)). n The molecular weight ranges from 500 Da to 10,000 kDa, 1 kDa to 1,000 kDa, or 10 kDa to 100 kDa. As used herein, “molecular weight” can refer to the relative average chain length of the bulk polymer and can be estimated or characterized in various ways, including gel permeation chromatography (GPC) and capillary viscometry. GPC molecular weight reports are number-average molecular weights (M).n ), rather than weight-average molecular weight (M). w Capillary viscosity determination provides molecular weight (M). v The estimated value of ) is the intrinsic viscosity of a dilute polymer solution determined using a specific set of concentration, temperature, and solvent conditions.

[0059] In some embodiments, the nanoparticles are made partly or entirely of one or more inorganic materials, such as clay (e.g., silicates) or other types of minerals (e.g., sulfides, oxides, halides, carbonates, sulfates, phosphates, apatite) or combinations thereof. Alternatively or in combination, the nanoparticles may be made partly or entirely of one or more metals, such as gold, silver, copper, platinum, palladium, ruthenium, or combinations thereof. Optionally, the nanoparticles may be made partly or entirely of carbon nanotubes (e.g., single-walled or multi-walled nanotubes), graphene, graphene oxide, or other ultrathin single crystals (including black phosphorus- and boron-based nanosheets).

[0060] In some embodiments, the nanoparticles are core-shell particles (also known as "corona particles"). Core-shell particles may have a core comprising or formed of a first material and a shell or corona comprising or formed of a different second material. For example, core-shell particles may comprise at least two polymers, such that the core is made of a first polymer and the shell or corona is made of a different second polymer. As another example, core-shell particles may comprise a single block copolymer, such that the core is made of a first block of the block copolymer and the shell or corona may be made of a second block of the block copolymer. In some embodiments, one or both components of the core-shell particles are non-polymer materials.

[0061] Core-shell particles can consist of two phases with different compositions, one (core or shell / crown) being hydrophobic and the other (core or shell / crown) being hydrophilic. Suitable hydrophobic components include polyamides (e.g., poly(amino acids)), polyesters (e.g., poly(lactic acid), poly(caprolactone)), polypropylene oxide, polystyrene, and combinations thereof. Suitable hydrophilic components include polysaccharides, proteins, polyamides (e.g., poly(amino acids)), naturally occurring polymers, synthetic polymers, and combinations thereof. Suitable block copolymers include combinations of polyethylene glycol and polyesters (e.g., PEG-PLA, poly(ethylene glycol)-block-poly(caprolactone) (PEG-PCL)) and combinations of polyethylene glycol and polypropylene glycol (e.g., poloxamers). In some embodiments, the core-shell particles are composed of amphiphilic polymers, including (1) one or more hydrophobic polymers selected from polyanhydrides, poly(orthoesters), polyesters, polyurethanes and / or copolymers thereof, and (2) one or more hydrophilic polymers selected from polysaccharides, proteins, poly(amino acids) and / or polyepoxides.

[0062] Alternatively, the nanoparticles can be homogeneous. Homogeneous nanoparticles can be formed uniformly from a single material or from multiple materials, which are not separated into different phases within the particle as in core-shell particles.

[0063] Nanoparticles can be prepared using techniques known in the art. The techniques used can depend on a variety of factors, including the materials used to form the nanoparticles, the desired size range of the resulting nanoparticles, and the suitability of the material to be encapsulated. Examples of suitable techniques include, but are not limited to, solvent evaporation, solvent removal, thermally melt microencapsulation, spray drying, phase inversion, polyelectrolyte condensation, monoemulsions and biemulsions (e.g., probe sonication), nanoparticle molding, and electrostatic self-assembly.

[0064] The concentration of nanoparticles in the PNP hydrogel can be varied to produce the desired hydrogel properties. In some embodiments, for example, the concentration of nanoparticles in the PNP hydrogel is 1 wt% to 15 wt%, 2 wt% to 12 wt%, 3 wt% to 10 wt%, 5 wt% to 8 wt%, 5 wt% to 15 wt%, 5 wt% to 10 wt%, 10 wt% to 15 wt%, or 10 wt% to 12 wt%. The concentration of nanoparticles in the PNP hydrogel can be about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, or about 15 wt%. In some embodiments, the concentration of nanoparticles in the PNP hydrogel can be greater than or equal to 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, 13 wt%, or 14 wt%. Alternatively or in combination, the concentration of nanoparticles in the PNP hydrogel can be less than or equal to 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, or 15 wt%.

[0065] 2. polymer

[0066] PNP hydrogels can be formed when nanoparticles are mixed with and interact with one or more polymers. As described herein, the shear-thinning and / or self-healing properties of PNP hydrogels can originate from reversible non-covalent interactions between the nanoparticles and the polymer chains. PNP hydrogels can contain a single type of polymer or a combination of multiple different polymers (e.g., two, three, four, five, or more different polymers). The polymers can be biodegradable and / or biocompatible. Polymers can include naturally occurring polymers, synthetic polymers, or derivatives or combinations thereof. Examples of naturally occurring polymers include polysaccharides (e.g., cellulose, alginate, collagen, chitosan, hyaluronic acid, starch, agarose, agar, xanthan gum), proteins (e.g., collagen, fibrin, albumin, zein, gelatin), and combinations thereof. Examples of synthetic polymers include polyacrylamide, poly(lactic acid), polyethylene glycol, polyethylene glycol-co-propylene glycol (PEO-PPO), polyacrylates (e.g., poly(2-hydroxyethyl methacrylate)), and combinations thereof. In some embodiments, the PNP hydrogel comprises a derivative of a naturally occurring polymer, such as a cellulose derivative. Examples of cellulose derivatives include hydroxypropyl methylcellulose (HPMC), hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), ethyl cellulose (EC), methylcellulose (MC), hydroxyethyl methylcellulose (HEMC), carboxymethyl cellulose (CMC), carboxymethyl ethyl cellulose (CMEC), and combinations thereof.

[0067] In some embodiments, the PNP hydrogel described herein comprises at least one polymer modified with a hydrophobic portion. The hydrophobic modification of the polymer can increase the interaction (αk) with each polymer nanoparticle. BThe modulus of the dynamic hydrogel is increased by modifying the energy related to T, thereby increasing the modulus of the hydrogel while having the same number of interactions per unit volume. Such modification can promote favorable interactions between the hydrophobic portion of the polymer chain and the hydrophobic core of the nanoparticle, thereby enhancing the adsorption energy of the polymer for the nanoparticle. The hydrophobic portion can contain multiple carbon atoms (e.g., 2 to 50 carbon atoms, 2 to 30 carbon atoms, or 2 to 18 carbon atoms) and can be a saturated or unsaturated molecule. Examples of hydrophobic moieties that may be used include, but are not limited to, alkyl moieties (e.g., C4 to C18 alkyl, such as butyl (-C4), hexyl (-C6), octyl (-C8), decyl (-C10), dodecyl (-C12), tetradecyl (-C14), pentadecyl (-C15), hexadecyl (-C16), heptadecanyl (-C17), octadecyl (-C18)), alkenyl moieties (e.g., oleyl, linoleyl), aryl moieties (e.g., phenyl, benzyl, pyrrole, naphthyl, anthracene), and cycloalkyl moieties (e.g., adamantyl, cyclohexyl, cholesterol). In some embodiments, the degree of polymer modification (e.g., the percentage of reactive groups on the polymer that have been functionalized with the hydrophobic moieties) is 1% to 50%, 5% to 30%, 5% to 25%, or 10% to 15%. For example, the degree of modification may be about 5%, 10%, 15%, 20%, or 25%.

[0068] The concentration of the polymer in the PNP hydrogel can be varied to produce desired hydrogel properties (e.g., stiffness, storage modulus, degradation rate). In some embodiments, for example, the concentration of the polymer in the PNP hydrogel is 0.25 wt% to 10 wt%, 0.5 wt% to 5 wt%, 0.5 wt% to 2 wt%, 1 wt% to 5 wt%, or 1 wt% to 2 wt%. The concentration of the polymer in the PNP hydrogel can be about 0.1 wt%, 0.25 wt%, 0.5 wt%, 0.75 wt%, 1 wt%, 1.25 wt%, 1.5 wt%, 1.75 wt%, 2 wt%, 2.25 wt%, 2.5 wt%, 2.75 wt%, 3 wt%, 3.5 wt%, 4 wt%, 4.5 wt%, or 5 wt%. In some embodiments, the polymer concentration in the PNP hydrogel can be greater than or equal to 0.25 wt%, 0.5 wt%, 0.75 wt%, 1 wt%, 1.25 wt%, 1.5 wt%, 1.75 wt%, 2 wt%, 2.25 wt%, 2.5 wt%, 2.75 wt%, 3 wt%, 3.5 wt%, 4 wt%, or 4.5 wt%. Alternatively or in combination, the polymer concentration in the PNP hydrogel can be less than or equal to 0.5 wt%, 0.75 wt%, 1 wt%, 1.25 wt%, 1.5 wt%, 1.75 wt%, 2 wt%, 2.25 wt%, 2.5 wt%, 2.75 wt%, 3 wt%, 3.5 wt%, 4 wt%, 4.5 wt%, or 5 wt%.

[0069] 3. Additional components

[0070] The PNP hydrogels described herein may optionally contain one or more additional components to promote gel formation and / or modify the properties of the hydrogel. For example, the PNP hydrogels described herein may contain at least one reinforcing compound that enhances the interaction between the polymer and the nanoparticles, for example, by providing a bridging non-covalent interaction between the polymer and the nanoparticles. In some embodiments, a portion of the reinforcing compound interacts non-covalently with the polymer, and a second portion of the reinforcing compound interacts non-covalently with the nanoparticles. Non-limiting examples of such interactions include ionic interactions (e.g., cation / anion interactions), electrostatic interactions, and hydrogen bonding interactions.

[0071] For example, in embodiments where the polymer is negatively charged at physiological pH (e.g., hyaluronic acid, carboxymethyl cellulose), cationic surfactants can be used to enhance the adsorption of anionic polymers to nanoparticles via electrostatic interactions. Examples of positively charged surfactants include hexadecyltrimethylammonium bromide (CTAB), hexadecyltrimethylammonium iodide, hexadecyltrimethylammonium fluoride, and hexadecyltrimethylammonium chloride. In contrast, in embodiments where the polymer is positively charged at physiological pH (e.g., chitosan, aminopolysaccharides, poly(lysine), cationic acrylate polymers, cationic vinyl polymers), anionic surfactants can be used to enhance the adsorption of cationic polymers to nanoparticles via electrostatic interactions. Examples of negatively charged surfactants include sodium dodecyl sulfate, sodium stearate, and charged fatty acid surfactants.

[0072] In some embodiments, molecular recognition between at least two compounds can provide an enhancing effect. For example, the adsorption of nanoparticles by polymers such as polysaccharides can be enhanced by an enhancing compound that contains a carbohydrate in a portion of the enhancing compound and includes a polymer tail that interacts with the nanoparticles.

[0073] The concentration of the reinforcing compound can be varied to produce the desired effect on hydrogel formation. In some embodiments, for example, the concentration of the reinforcing compound in the PNP hydrogel is 0.25 wt% to 10 wt%, 0.5 wt% to 5 wt%, 0.5 wt% to 2 wt%, 1 wt% to 5 wt%, or 1 wt% to 2 wt%. The concentration of the reinforcing compound in the PNP hydrogel can be about 0.1 wt%, 0.25 wt%, 0.5 wt%, 0.75 wt%, 1 wt%, 1.25 wt%, 1.5 wt%, 1.75 wt%, 2 wt%, 2.25 wt%, 2.5 wt%, 2.75 wt%, 3 wt%, 3.5 wt%, 4 wt%, 4.5 wt%, or 5 wt%. In some embodiments, the concentration of the reinforcing compound in the PNP hydrogel can be greater than or equal to 0.25 wt%, 0.5 wt%, 0.75 wt%, 1 wt%, 1.25 wt%, 1.5 wt%, 1.75 wt%, 2 wt%, 2.25 wt%, 2.5 wt%, 2.75 wt%, 3 wt%, 3.5 wt%, 4 wt%, or 4.5 wt%. Alternatively or in combination, the concentration of the reinforcing compound in the PNP hydrogel can be less than or equal to 0.5 wt%, 0.75 wt%, 1 wt%, 1.25 wt%, 1.5 wt%, 1.75 wt%, 2 wt%, 2.25 wt%, 2.5 wt%, 2.75 wt%, 3 wt%, 3.5 wt%, 4 wt%, 4.5 wt%, or 5 wt%. Optionally, the PNP hydrogel may not contain any reinforcing compound.

[0074] B. Hydrogel properties

[0075] The dynamic hydrogels described herein (e.g., the PNP hydrogels of Part IA) can exhibit favorable physical and biological properties that contribute to their efficacy as drug delivery platforms. The properties of the dynamic hydrogels described herein can be modulated in various ways, such as by changing the type of components used to form the hydrogel (e.g., polymers, nanoparticles, and / or additional components as previously described in Part IA; and / or therapeutic cargo carried by the hydrogel as described below in Part II), the concentration of the components, and / or the chemical functionality of the components. Therefore, the properties of the dynamic hydrogels described herein can be adapted to specific therapeutic applications, such as forming stable and / or persistent reservoirs when delivered in vivo, providing a desired release profile for the therapeutic cargo (e.g., short-term vs. long-term release), providing a desired release mechanism for the therapeutic cargo (e.g., diffusion-based vs. erosion-based release), compatibility with desired routes of administration (e.g., injection, infusion, spraying, application), biodegradability, biocompatibility, and / or allowing cell infiltration. Any reference in this document to the properties of dynamic hydrogels may refer to the properties of dynamic hydrogels without any therapeutic goods (e.g., PNP hydrogels consisting only of polymers and nanoparticles), the properties of dynamic hydrogels containing therapeutic goods (e.g., PNP hydrogels containing polymers, nanoparticles, and encapsulated therapeutic goods), or both, unless otherwise stated or apparent from the context.

[0076] The storage modulus (G') of a dynamic hydrogel can be related to the overall stiffness of the hydrogel, which in turn can define the timescale of hydrogel degradation (e.g., a hydrogel with a higher storage modulus can be stiffer and degrade more slowly than a hydrogel with a lower storage modulus). Therefore, in embodiments where the therapeutic cargo of the dynamic hydrogel is released primarily or entirely via an erosion-based mechanism, the release rate of the therapeutic cargo can be tuned by adjusting the storage modulus of the hydrogel (e.g., a higher storage modulus can result in a slower degradation rate and thus a slower release rate of the therapeutic cargo, while a lower storage modulus can result in a higher degradation rate and thus a faster release rate of the therapeutic cargo). For example, in embodiments where the dynamic hydrogel is a PNP hydrogel, the storage modulus of the PNP hydrogel can be increased or decreased by increasing or decreasing the polymer concentration and / or by increasing or decreasing the nanoparticle concentration. In some embodiments, the storage modulus of the dynamic hydrogel described herein is 1 Pa to 10,000 Pa, 1 Pa to 5000 Pa, 1 Pa to 2500 Pa, 1 Pa to 1000 Pa, 1 Pa to 500 Pa, 1 Pa to 200 Pa, 1 Pa to 10 Pa, 10 Pa to 10,000 Pa, 10 Pa to 5000 Pa, 10 Pa to 2500 Pa, 10 Pa to 1000 Pa, 10 Pa to 500 Pa, 10 Pa to 200 Pa, 10 Pa to 100 Pa, 10 Pa to 50 Pa, 50 Pa to 10,000 Pa, 50 Pa to 5000 Pa, 50 Pa to 2500 Pa, 50 Pa to 1000 Pa, 50 Pa to 5000 Pa, 50 Pa to 200 Pa, 50 Pa to 100 Pa, 100 Pa to 10,000 Pa, 100 Pa to 5000 Pa. Pa, 100 Pa to 2500 Pa, 100 Pa to 1000 Pa, 100 Pa to 500 Pa, 100 Pa to 200 Pa, 200 Pa to 10,000 Pa, 200 Pa to 5000 Pa, 200 Pa to 2500 Pa, 200 Pa to 1000 Pa, 200 Pa to 500 Pa, 500 Pa to 10,000 Pa, 500 Pa to 5000 Pa, 500 Pa to 2500 Pa, 500 Pa to 1000 Pa, 1000 Pa to 10,000 Pa, 1000 Pa to 5000 Pa, 1000 Pa to 2500 Pa, 2500 Pa to 10,000 Pa, 2500 Pa to 5000 Pa or 5000 Pa to 10,000 Pa.Storage modulus can be measured, for example, using an oscillating shear test in a parallel plate rheometer at an angular frequency of 10 rad / s, strain (e.g., 1% strain) in the linear viscoelastic region of the hydrogel, and a temperature of 25 °C.

[0077] Yield stress (τ) of dynamic hydrogels y This can be related to the hydrogel's ability to form and maintain a cohesive deposit in bulk (e.g., materials lacking yield stress can flow instead of forming a cohesive deposit). When subjected to stresses below the yield stress, the dynamic hydrogels of this paper may exhibit little or no flow. When subjected to stresses above the yield stress, the dynamic hydrogels can flow, corresponding to a significant decrease in viscosity observed (e.g., a decrease of at least one or two orders of magnitude). In embodiments where the dynamic hydrogel is a PNP hydrogel, the yield stress can be increased or decreased, respectively, by increasing or decreasing the nanoparticle concentration. In some embodiments, the yield stress of the dynamic hydrogel described herein is 0.1 Pa to 1000 Pa, 0.1 Pa to 500 Pa, 0.1 Pa to 200 Pa, 0.1 Pa to 100 Pa, 0.1 Pa to 50 Pa, 0.1 Pa to 20 Pa, 0.1 Pa to 10 Pa, 0.1 Pa to 1 Pa, 1 Pa to 1000 Pa, 1 Pa to 500 Pa, 1 Pa to 200 Pa, 1 Pa to 100 Pa, 1 Pa to 50 Pa, 1 Pa to 10 Pa, 10 Pa to 1000 Pa, 10 Pa to 500 Pa, 10 Pa to 200 Pa, 10 Pa to 100 Pa, 10 Pa to 50 Pa, 10 Pa to 20 Pa, 20 Pa to 1000 Pa, 20 Pa to 500 Pa, 20 Pa to 200 Pa, 20 Pa to 100 Pa, 20 Pa to 50 Pa, 50 Pa to 10 ... Pa to 500 Pa, 50 Pa to 200 Pa, 50 Pa to 100 Pa, 100 Pa to 500 Pa, 100 Pa to 200 Pa, 200 Pa to 1000 Pa, 200 Pa to 500 Pa, or 500 Pa to 1000 Pa. Yield stress can be measured, for example, in a parallel plate rheometer at a temperature of 25°C using a stress ramp or stress scan (e.g., 1 Pa to 100 Pa, or 1 Pa to 1000 Pa) to determine the stress at which the hydrogel exhibits a decrease in viscosity.

[0078] The tan δ (the ratio of loss modulus (G”) to storage modulus (G’) of a dynamic hydrogel (tan (δ) = G” / G’)) can describe the overall viscoelasticity of the hydrogel (e.g., a lower tan δ value corresponds to more solid-like behavior, and a higher tan δ value corresponds to more liquid-like behavior) and can be correlated with the degradation rate of the hydrogel. In some embodiments, the tan δ of the dynamic hydrogel described herein is less than or equal to 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1. tan δ can be from 0.1 to 1, 0.1 to 0.5, 0.1 to 0.3, 0.2 to 1, 0.2 to 0.5, or 0.5 to 1. tan δ can be measured, for example, using an oscillating shear test in a parallel plate rheometer at an angular frequency of 10 rad / s, strain (e.g., 1% strain) within the linear viscoelastic region of the hydrogel, and a temperature of 25°C.

[0079] In some embodiments, the dynamic hydrogels described herein exhibit shear-thinning behavior because the viscosity of the dynamic hydrogel decreases with increasing shear rate and / or shear stress. Shear-thinning behavior can be advantageous, for example, allowing the dynamic hydrogel to be applied via injection. In some embodiments, the viscosity of the gel is in the range of 0.1 s⁻¹. -1 up to 1000 s -1 The shear rate decreases with increasing shear rate, for example, as observed on an oscillating rheometer (e.g., a parallel plate rheometer) at 25°C. In some embodiments, at 1000 s... -1At the shear rates described in this paper, the viscosities of the dynamic hydrogels were 10 mPa⁻² to 2000 mPa⁻², 10 mPa⁻² to 1000 mPa⁻², 10 mPa⁻² to 500 mPa⁻², 10 mPa⁻² to 200 mPa⁻², 10 mPa⁻² to 100 mPa⁻², 10 mPa⁻² to 50 mPa⁻², 50 mPa⁻² to 2000 mPa⁻², 50 mPa⁻² to 1000 mPa⁻², 50 mPa⁻² to 2000 mPa⁻², 50 mPa⁻² to 1000 mPa⁻², 100 mPa⁻² to 2 ... Viscosities range from 200 mPa-s to 2000 mPa-s, 200 mPa-s to 1000 mPa-s, 200 mPa-s to 500 mPa-s, 500 mPa-s to 2000 mPa-s, 500 mPa-s to 1000 mPa-s, or 1000 mPa-s to 2000 mPa-s. Viscosities can be measured, for example, at a temperature of 25°C using steady-state shear measurements in a parallel plate rheometer.

[0080] In some embodiments, the dynamic hydrogels of this invention exhibit self-healing behavior. Self-healing can refer to a process in which a gel exhibiting reduced flow resistance when subjected to external stress recovers some or all of its stiffness and / or strength after the external stress is removed. Self-healing behavior can be advantageous, for example, allowing the dynamic hydrogel to form a cohesive reservoir and / or restrict burst release after injection application. In some embodiments, the dynamic hydrogels of the present invention cease flow and recover their mechanical properties within no more than 5 seconds, 10 seconds, 30 seconds, 1 minute, 2 minutes, 3 minutes, 5 minutes, or 10 minutes after the external stress is removed. Optionally, the modulus and / or viscosity of the dynamic hydrogel can be measured on an oscillating rheometer using step strain measurements (with strains of 0.5% and 500%) or step shear measurements (with strains of 0.1 s⁻¹). -1 and 100 s -1 The shear rate is restored to at least 90% of its initial value before the application of external stress within 5 minutes.

[0081] In some implementations, the dynamic hydrogels described herein exhibit viscoelastic behavior because the storage modulus (G') of the hydrogel dominates at some point compared to the loss modulus (G”), as observed, for example, in oscillatory frequency sweep measurements from 0.1 rad / s to 100 rad / s on an oscillatory rheometer in the linear viscoelastic region; however, the hydrogel exhibits complete stress relaxation within 15 minutes after the application of a constant strain of 500%.

[0082] In some embodiments, the dynamic hydrogels described herein are biocompatible. Biocompatible materials can be those whose metabolites or degradation products are generally non-toxic to the subject at concentrations caused by degradation of the applied material and do not cause any significant adverse effects on the subject. Biocompatible materials can be those that do not elicit a significant inflammatory or immune response when applied to the subject.

[0083] In some embodiments, the dynamic hydrogel described herein is biodegradable. Biodegradable materials can be materials that degrade or corrode under physiological conditions into smaller units or chemical substances that can be metabolized, eliminated, or excreted by the subject. For example, upon administration into the body, the dynamic hydrogel can dissolve with the dissociation of non-covalent bonds. The degradation rate of the dynamic hydrogel can vary as desired, for example, depending on the desired release profile of the therapeutic cargo. In some embodiments, following in vivo administration, the dynamic hydrogel is designed to remain at the application site (e.g., as a cohesive reservoir) for at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 21 days, 28 days, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 9 months, or 12 months. Optionally or in combination, the dynamic hydrogel described herein may remain at the application site for no more than 12 months, 9 months, 6 months, 5 months, 4 months, 3 months, 2 months, 1 month, 28 days, 21 days, 14 days, 13 days, 12 days, 11 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day.

[0084] II. Compositions and related methods for delivering therapeutic peptides

[0085] In some embodiments, this technology provides compositions for delivering therapeutic peptides of a disease or condition to a subject. The composition may comprise a dynamic hydrogel and at least one therapeutic peptide carried by the dynamic hydrogel. The dynamic hydrogel may encapsulate the therapeutic peptide and provide sustained, controlled release of the therapeutic peptide when the composition is applied to the subject. In some embodiments, the dynamic hydrogel exhibits shear-thinning behavior that allows for easy administration via injection, and self-healing behavior that allows for the formation of a cohesive reservoir for delivering the therapeutic peptide over an extended therapeutic period. For example, Figure 1AThis is a schematic diagram of a PNP hydrogel prepared by mixing hydrophobically modified HPMC with PEG-PLA nanoparticles. This hydrogel allows for easy encapsulation of therapeutic peptides, and... Figure 1B This is a schematic diagram illustrating how a local reservoir is formed in the subcutaneous space after subcutaneous injection of PNP hydrogel, thereby providing a tunable platform for the sustained release of therapeutic peptides.

[0086] A. Therapeutic peptides

[0087] 1. Incretin mimics

[0088] In some implementations, the therapeutic peptide is an incretin mimic. An incretin mimic is a compound that mimics the activity of incretin hormones. Incretin hormones are peptides secreted by the intestine in response to nutrient intake. The main incretin hormones are GLP-1 and gastric inhibitory peptide (GIP). GLP-1 contributes to the regulation of glucose homeostasis in the body through its interaction with its receptor. GLP-1 is secreted from L-cells in the intestine in response to nutrients and lowers blood glucose by stimulating insulin in a glucose-dependent manner and inhibiting glucagon secretion, thereby reducing the risk of hypoglycemia. Furthermore, GLP-1 is also a neurotransmitter synthesized by preglucagonogenamic neurons in the brain and reduces energy intake via central pathways by influencing satiety, hunger, and reward-related measures, leading to weight loss. The metabolic effects of GLP-1 include glucose-dependent stimulation of insulin secretion, inhibition of glucagon secretion, inhibition of food intake, reduction of gastric emptying, and increased natriuretic and diuretic effects. GLP-1 has also been shown to influence learning, memory, reward behavior, and palatability, and exhibits neuroprotective, cardioprotective, and anti-inflammatory effects. However, the therapeutic application of natural GLP-1 is limited by its short in vivo half-life (approximately 2 to 3 minutes) and inactivation by dipeptidyl peptidase 4 (DPP4). GIP is secreted by enteroendocrine K cells in response to nutrients and also exhibits insulinotropic effects via binding to GIP receptors. However, unlike GLP-1, GIP stimulates glucagon secretion. GIP also affects appetite, fat accumulation, memory, and bone formation. Natural GIP also exhibits a short half-life (approximately 4 to 5 minutes) and is inactivated by DPP4.

[0089] In some implementations, the incretin mimic is a GLP-1 RA. GLP-1 RAs (also known as “GLP-1 analogs”) are a class of drugs that interact with the GLP-1 receptor and exhibit structural similarity to native GLP-1, but with modifications to prolong their in vivo half-life and thus provide improved bioavailability. GLP-1 RAs can be classified as short-acting or long-acting compounds. Short-acting GLP-1 RAs are made resistant to DPP4 cleavage by altering the second and third amino acids at the N-terminus, but still undergo renal clearance, and therefore typically have a half-life of approximately 2 to 5 hours. Examples of short-acting GLP-1 RAs include exenatide and lixisenatide. Long-acting GLP-1 RAs implement mechanisms that reduce renal clearance, such as fatty acid acylation to promote binding to serum albumin or conjugation to larger molecules / components, and can therefore have a half-life of 12 hours to several days. Examples of long-acting GLP-1 RAs include liraglutide (C16 fatty acid monoacylation), smegglutide (C18 fatty acid diacidation), telpoglutide (C20 fatty acid diacidation), retatrutide (C20 fatty acid diacidation), albiglutide (conjugated to albumin), dulaglutide (conjugated to the Fc fragment of IgG), and exenatide-LAR (long-acting release) (conjugated to biodegradable polymer microspheres).

[0090] GLP-1 RAs may include peptides that bind to the GLP-1 receptor. The peptides may be analogs of natural GLP-1 peptides, such as endogenous human GLP-1 peptides (e.g., GLP-1 (7-36) or GLP-1 (7-37)). For example, the peptides of GLP-1 RAs may contain a sequence having at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or 100% sequence identity with either SEQ ID NO: 1 or SEQ ID NO: 2. The peptides may be produced in suitable host cells via recombinant DNA technology, in cell-free systems, or synthesized via solid-phase synthesis.

[0091] Table 1: GLP-1 peptides

[0092] In some implementations, GLP-1 RAs are monoreceptor agonists that specifically bind to the GLP-1 receptor. Examples of GLP-1 monoreceptor agonists include exenatide, exenatide-LAR, lixinatide, liraglutide, smegglutide, abiglutide, duraglutide, efpeglenatide, and ecnoglutide. As previously mentioned, the peptide of a GLP-1 monoreceptor agonist can be an analogue of a natural GLP-1 peptide.

[0093] In some embodiments, the GLP-1 RA is a dual-receptor agonist that binds to the GLP-1 receptor and an additional receptor. The additional receptor can be any of the following receptors: glucagon receptor, GIP receptor, cholecystokinin receptor, xenin receptor, secretin receptor, neuropeptide Y receptor, or neurotensin receptor. For example, the GLP-1 RA can be a GLP-1 / glucagon dual-receptor agonist that binds to both the GLP-1 receptor and the glucagon receptor. Examples of GLP-1 / glucagon dual-receptor agonists include efinopegdutide, cotadutide, mazdutide, and BI 45690. In such embodiments, the peptide of the GLP-1 / glucagon dual-receptor agonist can be an analogue of oxyntomodulin, which is an intestinal hormone that activates both the GLP-1 receptor and the glucagon receptor. As another example, a GLP-1 RA can be a GLP-1 / GIP dual-receptor agonist that binds to both the GLP-1 receptor and the GIP receptor. Examples of GLP-1 / GIP dual-receptor agonists include telpolide, LY3493269, VK2735, CT-868, and AMG133. In such embodiments, the peptide of the GLP-1 / GIP dual-receptor agonist can be a GIP analog that has high sequence similarity to GLP-1 in the N-terminal portion of the peptide.

[0094] In some implementations, the GLP-1 RA is a triadic receptor agonist that binds to the GLP-1 receptor and two additional receptors. The additional receptors can be any two of the following receptors: glucagon receptor, GIP receptor, cholecystokinin receptor, xenin receptor, secretin receptor, neuropeptide Y receptor, or neurotensin receptor. For example, the GLP-1 RA can be a GLP-1 / glucagon / GIP triadic receptor agonist that binds to the GLP-1 receptor, glucagon receptor, and GIP receptor. Examples of GLP-1 / glucagon / GIP triadic receptor agonists include retaglutide.

[0095] The peptide of GLP-1 RA can be linked to at least one substituent (also referred to herein as a “side chain”). The substituent can prolong the half-life of the peptide in vivo, for example, through binding to serum albumin. Optionally, the substituent can provide other beneficial effects, such as enhanced solubility, promoted cellular uptake, and / or reduced immunogenicity. The substituent of GLP-1 RA can be linked to the peptide of GLP-1 RA via any suitable mechanism, such as acylation, alkylation, esterification, amide formation, coupling with cysteine ​​residues, and / or other conjugation chemistry mechanisms known to those skilled in the art. For example, the substituent can be covalently linked to the peptide via an amide bond between the carboxyl group of the substituent and the amino group of the peptide. The amino group of the peptide can be the N-terminal amino group of the peptide or a side chain amino group of an amino acid residue of the peptide (e.g., the amino group of a lysine residue of the peptide). The substituent can be directly linked to the peptide or indirectly linked to the peptide via a linker, also referred to herein as a spacer. For example, the substituent can be linked to the peptide via an amide bond between the carboxyl group of the linker and the amino group of an amino acid residue of the peptide. The linker can be any suitable linker known to those skilled in the art, such as peptide linkers (e.g., γ-glutamic acid linkers), hydrophilic spacers (e.g., 8-amino-3,6-dioxanoic acid), hydrophobic spacers, or combinations thereof.

[0096] For example, GLP-1 RA can be an acylated peptide linked to a lipophilic substituent having multiple carbon atoms (e.g., at least 10, 15, 20, 25, 30, 35, or 40 carbon atoms). In some embodiments, the lipophilic substituent is an acyl group of a fatty acid, such as a straight-chain fatty acid or a branched-chain fatty acid. The fatty acid can be a fatty monoacid, such as an aliphatic monocarboxylic acid having 4 to 38 carbon atoms, which can be saturated or unsaturated. The fatty acid can be a fatty diacid, such as an aliphatic dicarboxylic acid having 4 to 38 carbon atoms, which can be saturated or unsaturated. Fatty acids can be C4-C38 fatty acids, such as C4 fatty acids, C6 fatty acids, C8 fatty acids, C10 fatty acids, C12 fatty acids, C14 fatty acids, C15 fatty acids, C16 fatty acids, C17 fatty acids, C18 fatty acids, C20 fatty acids, C22 fatty acids, C24 fatty acids, C26 fatty acids, C28 fatty acids, C30 fatty acids, C32 fatty acids, C34 fatty acids, C36 fatty acids, or C38 fatty acids.

[0097] In some embodiments, the lipophilic substituent is of the formula CH3(CH2). n An acyl group of CO-, where n is an integer from 4 to 38 or from 4 to 24, for example CH3(CH2)4CO-, CH3(CH2)6CO-, CH3(CH2)8CO-, CH3(CH2) 10CO-, CH3(CH2) 12 CO-, CH3(CH2) 14 CO-, CH3(CH2) 16 CO-, CH3(CH2) 18 CO-, CH3(CH2) 20 CO-, CH3(CH2) 22 CO- or CH3(CH2) 24 CO-.

[0098] In some implementations, the lipophilic substituent is of the formula HOOC(CH2). n An acyl group of CO-, where n is an integer from 4 to 38 or from 4 to 24, for example, HOOC(CH2). 14 CO-, HOOC(CH2) 16 CO-, HOOC(CH2) 18 CO-, HOOC(CH2) 20 CO- or HOOC(CH2) 22 CO-.

[0099] In some embodiments, the lipophilic substituent is an acyl group of a straight-chain or branched alkane α,ω-dicarboxylic acid.

[0100] In some implementations, the lipophilic substituent is of the formula HOOC(CH2). n An acyl group of CO-, where n is an integer from 4 to 38 or from 4 to 24, for example, HOOC(CH2). 16 CO-, HOOC(CH2) 18 CO-, HOOC(CH2) 20 CO- or HOOC(CH2) 22 CO.

[0101] In some embodiments, the lipophilic substituent is of the formula CH3(CH2). n The acyl group of CO-NHCH(COOH)(CH2)2CO-, where n is an integer from 10 to 24.

[0102] In some embodiments, the lipophilic substituent is of the formula CH3(CH2). n The acyl group of CO-NHCH((CH2)2COOH)CO-, where n is an integer from 8 to 24.

[0103] In some embodiments, the lipophilic substituent is a substituent with the formula COOH(CH2). n The acyl group of CO-, where n is an integer from 8 to 24.

[0104] In some embodiments, the lipophilic substituent is of the formula -NHCH(COOH)(CH2)4NH-CO(CH2). n The acyl group of CH3, where n is an integer from 8 to 18.

[0105] Lipophilic substituents can be linked to peptides of GLP-1 RA via linkers, as described herein. In some embodiments, the lipophilic substituents interact with components of the dynamic hydrogel to facilitate the encapsulation and controlled release of GLP-1 RA by the dynamic hydrogel, as further described below.

[0106] In some embodiments, the composition of this technology comprises a dynamic hydrogel and an incretin mimic encapsulated by the dynamic hydrogel. The incretin mimic may be one or more of a GLP-1 RA, such as exenatide, exenatide-LAR, lixinatide, liraglutide, smegglutide, abiglutide, dulaglutide, ipegnatide, enogglutide, ifenoopeptide, cotodextrin, mastodextrin, BI 45690, telpolide, LY3493269, VK2735, CT-868, AMG133, or retaglutide. In some embodiments, the incretin mimic is a GLP-1 RA, which is an acylated peptide, such as one or more of liraglutide, smegglutide, enogglutide, cotodextrin, mastodextrin, telpolide, or retaglutide. Additional examples of GLP-1 RAs and incretin mimics are provided in International Publications WO 2005 / 027978 and WO 2014 / 005858 (the disclosures of each of which are incorporated herein by reference in their entirety). Optionally, the composition may comprise a combination of two or more different incretin mimics, such as two or more of any of the incretin mimics disclosed herein.

[0107] Optionally, in addition to incretin mimics, the compositions herein may also comprise other therapeutic goods carried by a dynamic hydrogel. Other therapeutic goods may include one or more therapeutic agents that produce the desired therapeutic effect, such as small molecule drugs, peptides, proteins, polysaccharides, nucleic acids, cells, or combinations thereof. In some embodiments, the therapeutic agent works synergistically with the incretin mimic to treat diseases or conditions, such as antidiabetic agents, anti-obesity agents, appetite suppressants, and / or antihypertensive agents. Examples of such therapeutic agents include alpha-glucosidase inhibitors, amylin analogs (e.g., canagliptin, biguanides, DPP4 inhibitors, glucagon antagonists, insulin and insulin analogs (e.g., degludec insulin, detemir insulin, icodec insulin, glargine insulin), meglitinides, SGLT-2 inhibitors, sulfonylureas, thiazolidinediones, and combinations thereof. Such therapeutic agents can be encapsulated in a dynamic hydrogel via physical encapsulation, interaction with hydrogel components (e.g., hydrophobic interactions), or a suitable combination thereof. For example, a therapeutic agent precipitated at physiological pH (e.g., glargine insulin) can be physically encapsulated within a hydrogel. Optionally, the therapeutic agent can be administered separately from the incretin mimic via any suitable route of administration (e.g., parenteral or non-parenteral administration).

[0108] 2. Acyl peptide

[0109] Although some embodiments of the compositions described herein are in combination with incretin mimics such as GLP-1 RAs, this is not intended to be limiting, and the compositions of this technology can be used to deliver other types of therapeutic peptides, such as acylated peptides. Acylation of peptides having lipophilic substituents (e.g., fatty acid moieties) can produce improved pharmacokinetics via binding to serum albumin compared to native peptides, while maintaining the activity of the native peptide. Other beneficial effects of acylation may include enhanced solubility, improved cellular uptake, and / or reduced immunogenicity.

[0110] In some embodiments, the composition of this technology comprises a dynamic hydrogel and at least one acylated peptide encapsulated by the dynamic hydrogel. The acylated peptide may comprise a peptide that exhibits a therapeutic effect when administered to a subject, and at least one substituent linked to the peptide via acylation. The peptide may be a natural peptide (e.g., having 100% sequence identity with an endogenous human peptide) or an analogue with one or more modifications relative to a natural peptide (e.g., having less than 100% sequence identity with an endogenous human peptide). For example, the natural peptide sequence may be modified by substituting one or more amino acids (e.g., with natural or non-natural amino acids), adding one or more amino acids (e.g., natural or non-natural amino acids), deleting one or more amino acids, or suitable combinations thereof. The peptide may be produced in suitable host cells via recombinant DNA technology, may be produced in a cell-free system, or may be synthesized via solid-phase synthesis.

[0111] The lipophilic substituent of the acylated peptide can have multiple carbon atoms, such as at least 10, 15, 20, 25, 30, 35, or 40 carbon atoms. In some embodiments, the lipophilic substituent is an acyl group of a fatty acid, such as a straight-chain fatty acid or a branched-chain fatty acid. The fatty acid can be a fatty monoacid, such as an aliphatic monocarboxylic acid having 4 to 38 carbon atoms, which can be saturated or unsaturated. The fatty acid can be a fatty diacid, such as an aliphatic dicarboxylic acid having 4 to 38 carbon atoms, which can be saturated or unsaturated. Fatty acids can be C4-C38 fatty acids, such as C4 fatty acids, C6 fatty acids, C8 fatty acids, C10 fatty acids, C12 fatty acids, C14 fatty acids, C15 fatty acids, C16 fatty acids, C17 fatty acids, C18 fatty acids, C20 fatty acids, C22 fatty acids, C24 fatty acids, C26 fatty acids, C28 fatty acids, C30 fatty acids, C32 fatty acids, C34 fatty acids, C36 fatty acids, or C38 fatty acids.

[0112] In some embodiments, the lipophilic substituent is of the formula CH3(CH2). n An acyl group of CO-, where n is an integer from 4 to 38 or from 4 to 24, for example CH3(CH2)4CO-, CH3(CH2)6CO-, CH3(CH2)8CO-, CH3(CH2) 10 CO-, CH3(CH2) 12 CO-, CH3(CH2) 14 CO-, CH3(CH2) 16 CO-, CH3(CH2) 18 CO-, CH3(CH2) 20 CO-, CH3(CH2) 22 CO- or CH3(CH2) 24CO-.

[0113] In some implementations, the lipophilic substituent is of the formula HOOC(CH2). n An acyl group of CO-, where n is an integer from 4 to 38 or from 4 to 24, for example, HOOC(CH2). 14 CO-, HOOC(CH2) 16 CO-, HOOC(CH2) 18 CO-, HOOC(CH2) 20 CO- or HOOC(CH2) 22 CO-.

[0114] In some embodiments, the lipophilic substituent is an acyl group of a straight-chain or branched alkane α,ω-dicarboxylic acid.

[0115] In some implementations, the lipophilic substituent is of the formula HOOC(CH2). n An acyl group of CO-, where n is an integer from 4 to 38, or from 4 to 24, for example, HOOC(CH2). 16 CO-, HOOC(CH2) 18 CO-, HOOC(CH2) 20 CO- or HOOC(CH2) 22 CO.

[0116] In some embodiments, the lipophilic substituent is of the formula CH3(CH2). n The acyl group of CO-NHCH(COOH)(CH2)2CO-, where n is an integer from 10 to 24.

[0117] In some embodiments, the lipophilic substituent is of the formula CH3(CH2). n The acyl group of CO-NHCH((CH2)2COOH)CO-, where n is an integer from 8 to 24.

[0118] In some embodiments, the lipophilic substituent is a substituent with the formula COOH(CH2). n The acyl group of CO-, where n is an integer from 8 to 24.

[0119] In some embodiments, the lipophilic substituent is of the formula -NHCH(COOH)(CH2)4NH-CO(CH2). n The acyl group of CH3, where n is an integer from 8 to 18.

[0120] In some implementations, lipophilic substituents interact with components of the dynamic hydrogel to facilitate the encapsulation and controlled release of acylated peptides by the dynamic hydrogel, as further described below.

[0121] In some embodiments, the lipophilic substituent is covalently linked to the peptide via an amide bond between the carboxyl group of the substituent and the amino group of the peptide. The amino group of the peptide can be the N-terminal amino group of the peptide or a side-chain amino group of an amino acid residue of the peptide (e.g., the amino group of a lysine residue). The lipophilic substituent can be directly linked to the peptide or indirectly linked to the peptide via a linker. For example, the lipophilic substituent can be linked to the peptide via an amide bond between the carboxyl group of the linker and the amino group of an amino acid residue of the peptide. The linker can be any suitable linker known to those skilled in the art, such as peptide linkers (e.g., amino acid, γ-glutamic acid linkers), hydrophilic spacers (e.g., PEG, 8-amino-3,6-dioxanoic acid (OEG)), hydrophobic spacers, or combinations thereof.

[0122] In some embodiments, the acylated peptide is an acylated analog of a proglucagon-derived peptide. Proglucagon-derived peptides are a family of differentially processed peptides derived from a common pro-hormone (proglucagon) and include glucagon, GLP-1, glucagon-like peptide-2 (GLP-2), gastrin (OXM), enteroglucagon, enteroglucagon-associated pancreatic peptide (GRPP), intercalation peptide-1 (IP-1), intercalation peptide-2 (IP-2), and major proglucagon fragment (MPGF). These peptides exhibit a variety of physiological functions, including metabolism, energy regulation, cardioprotection, bone health, kidney function, liver function, and cognition. Acylated analogs of proglucagon peptides may include the peptide and lipophilic substituents linked to the peptide as described herein, said peptide comprising a sequence having at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or 100% sequence identity with the natural proglucagon peptide.

[0123] In some embodiments, the acylated peptide is an acylated amylin analogue, such as canagliflozin. Amylin is a hormone secreted by pancreatic β-cells in response to nutrient intake. Amylin signaling plays a role in glycemic regulation by delaying gastric emptying, inhibiting food intake, and suppressing meal-related glucagon secretion, and is therefore complementary to the activity of incretin hormones. Acylated amylin analogues may comprise a peptide and a lipophilic substituent linked to the peptide as described herein, said peptide comprising a sequence having at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or 100% sequence identity with the native amylin peptide.

[0124] In some embodiments, the acylated peptide is an acylated insulin analog, such as insulin degludec, insulin detemir, or insulin icotin. The acylated insulin analog can be co-delivered with an incretin mimic for the treatment of diabetes. The acylated insulin analog may include a peptide and lipophilic substituents linked to the peptide as described herein, said peptide comprising a sequence having at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or 100% sequence identity with the native insulin peptide.

[0125] B. Compositions and methods

[0126] In some embodiments, this technology provides a composition comprising a dynamic hydrogel and one or more therapeutic peptides (e.g., incretin mimics and / or acylated peptides) encapsulated by the dynamic hydrogel. The dynamic hydrogel carrying the therapeutic peptide can be any of the dynamic hydrogels described in Part I above. For example, the dynamic hydrogel can be a PNP hydrogel composed of a polymer and multiple nanoparticles that interact non-covalently with each other, as previously discussed in Part IA. The dynamic hydrogel can exhibit shear-thinning, self-healing, and / or viscoelastic properties resulting from non-covalent supramolecular interactions between the hydrogel components, as described above in Part IB.

[0127] The composition may contain any suitable amount of a therapeutic peptide for providing the desired therapeutic effect. For example, the composition may contain at least 1 mg, 2 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, or 400 mg of a therapeutic peptide. Optionally or in combination, the composition may contain a therapeutic peptide in amounts not exceeding 500 mg, 400 mg, 300 mg, 275 mg, 250 mg, 225 mg, 200 mg, 175 mg, 150 mg, 125 mg, 100 mg, 90 mg, 80 mg, 70 mg, 60 mg, 50 mg, 45 mg, 40 mg, 35 mg, 30 mg, 25 mg, 20 mg, 15 mg, 10 mg, 5 mg, or 2 mg. The amount of therapeutic peptide in the composition can be 1 mg to 500 mg, 1 mg to 250 mg, 1 mg to 150 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 40 mg, 1 mg to 30 mg, 1 mg to 20 mg, 1 mg to 10 mg, 10 mg to 500 mg, 10 mg to 250 mg, 10 mg to 150 mg, 10 mg to 100 mg, 10 mg to 50 mg, 10 mg to 40 mg, 10 mg to 30 mg, 10 mg to 20 mg, 20 mg to 500 mg, 20 mg to 250 mg, 20 mg to 150 mg, 20 mg to 100 mg, 20 mg to 50 mg, 20 mg to 40 mg, 20 mg to 30 mg, 30 mg to 500 mg, 30 mg to 250 mg, 30 mg to 150 mg, 30 mg to 100 mg, 30 mg to 50 mg. mg, 30 mg to 40 mg, 40 mg to 500 mg, 40 mg to 250 mg, 40 mg to 150 mg, 40 mg to 100 mg, 40 mg to 50 mg, 50 mg to 500 mg, 50 mg to 250 mg, 50 mg to 150 mg, 50 mg to 100 mg, 100 mg to 500 mg, 100 mg to 250 mg, 100 mg to 150 mg, 150 mg to 500 mg, 150 mg to 250 mg or 250 mg to 500 mg.The amount of therapeutic peptide in the composition may be approximately 1 mg, 2 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 400 mg, or 500 mg.

[0128] In some embodiments, the therapeutic peptide is present in the composition at a concentration of at least 0.5 mg / mL, 1 mg / mL, 1.5 mg / mL, 2 mg / mL, 5 mg / mL, 10 mg / mL, 20 mg / mL, 50 mg / mL, 100 mg / mL, 120 mg / mL, or 150 mg / mL. Optionally or in combination, the concentration of the therapeutic peptide in the composition does not exceed 200 mg / mL, 150 mg / mL, 120 mg / mL, 100 mg / mL, 50 mg / mL, 20 mg / mL, 10 mg / mL, 5 mg / mL, 2 mg / mL, 1.5 mg / mL, 1 mg / mL, or 0.5 mg / mL. The concentration of the therapeutic peptide in the composition can be from 0.5 mg / mL to 200 mg / mL, 0.5 mg / mL to 150 mg / mL, 0.5 mg / mL to 120 mg / mL, 0.5 mg / mL to 100 mg / mL, 0.5 mg / mL to 50 mg / mL, 0.5 mg / mL to 20 mg / mL, 0.5 mg / mL to 10 mg / mL, 0.5 mg / mL to 5 mg / mL, 0.5 mg / mL to 2 mg / mL, 0.5 mg / mL to 1.5 mg / mL, 0.5 mg / mL to 1 mg / mL, 1 mg / mL to 200 mg / mL, 1 mg / mL to 150 mg / mL, 1 mg / mL to 120 mg / mL, 1 mg / mL to 100 mg / mL, 1 mg / mL to 50 mg / mL, 1 mg / mL to 20 mg / mL, 1 mg / mL to 10 mg / mL, 1 mg / mL to 50 mg / mL. mg / mL, 1 mg / mL to 2 mg / mL, 1 mg / mL to 1 mg / mL to 1.5 mg / mL, 1.5 mg / mL to 200 mg / mL, 1.5 mg / mL to 150 mg / mL, 1.5 mg / mL to 120 mg / mL, 1.5 mg / mL to 100 mg / mL, 1.5 mg / mL to 50 mg / mL, 1.5 mg / mL to 20 mg / mL, 1.5 mg / mL to 10 mg / mL, 1.5 mg / mL to 5mg / mL, 1.5 mg / mL to 2 mg / mL, 2 mg / mL to 200 mg / mL, 2 mg / mL to 150 mg / mL, 2 mg / mL to 120 mg / mL, 2 mg / mL to 100 mg / mL, 2 mg / mL to 50 mg / mL, 2 mg / mL to 20 mg / mL, 2 mg / mL to 10 mg / mL, 2 mg / mL to 5 mg / mL, 5 mg / mL to 200 mg / mL, 5 mg / mL to 150 mg / mL, 5 mg / mL to 120 mg / mL, 5 mg / mL to 100 mg / mL, 5 mg / mL to 50 mg / mL, 5 mg / mL to 20 mg / mL, 5 mg / mL to 10 mg / mL, 10 mg / mL to 200 mg / mL, 10 mg / mL to 150 mg / mL, 10 mg / mL to 120 mg / mL, 10 mg / mL to 100 mg / mL, 10 mg / mL to 50 mg / mL, 10 mg / mL to 20 mg / mL, 20 mg / mL to 200 mg / mL, 20 mg / mL to 150 mg / mL, 20 mg / mL to 120 mg / mL, 20 mg / mL to 100 mg / mL, 20 mg / mL to 50 mg / mL, 50 mg / mL to 200mg / mL, 50 mg / mL to 150 mg / mL, 50 mg / mL to 120 mg / mL, 50 mg / mL to 100 mg / mL or 100 mg / mL to 200 mg / mL. .

[0129] In some implementations, the size of the therapeutic peptide is smaller than the pore size of the dynamic hydrogel. For example, the pore size of the dynamic hydrogel can be greater than or equal to 2 nm, 2.5 nm, 3 nm, 3.5 nm, 4 nm, 4.5 nm, or 5 nm; while the size of the therapeutic peptide (e.g., hydrodynamic diameter) can be less than 2 nm, 1.5 nm, 1 nm, or 0.5 nm. Therefore, physical encapsulation of the therapeutic peptide through the hydrogel network may be ineffective for achieving controlled release, as the therapeutic peptide may be able to diffuse freely out of the dynamic hydrogel, resulting in an uncontrolled burst release in vivo. For example, physical encapsulation may be insufficient to control the release of therapeutic peptides with molecular weights less than or equal to 5 kDa, 4.5 kDa, 4 kDa, or 3.5 kDa.

[0130] In such embodiments, the dynamic hydrogel may comprise at least one component that binds to the therapeutic peptide to control the release of the therapeutic peptide from the dynamic hydrogel. The interaction can be non-covalent, such as a hydrophobic interaction. For example, in an embodiment where the therapeutic peptide is acylated with a fatty acid, the lipids in the fatty acid can interact with one or more hydrophobic components of the dynamic hydrogel (e.g., for PNP hydrogels, hydrophobic surfaces of nanoparticles, and / or hydrophobic portions on polymer chains). These hydrophobic interactions may cause the therapeutic peptide to adhere to the hydrophobic components of the dynamic hydrogel, thereby inhibiting the uncontrolled diffusion of the therapeutic peptide out of the dynamic hydrogel. In such embodiments, the therapeutic peptide can be released from the dynamic hydrogel primarily or entirely via erosion of the dynamic hydrogel in vivo. Therefore, the release kinetics of the therapeutic peptide can be modulated by adjusting the degradation rate of the dynamic hydrogel (e.g., by controlling the storage modulus of the dynamic hydrogel).

[0131] In some cases, therapeutic peptides can exhibit aggregation behavior that may interfere with the binding of the therapeutic peptide to a dynamic hydrogel, resulting in the unintended diffusion and release of a significant portion of the "free" peptide from the hydrogel within a short period. For example, semaglutide has been shown to form dimer substances. When dimerized, the fatty acid side chains of semaglutide may not be able to interact hydrophobically with the hydrogel, but the semaglutide dimer may still be too small to be physically encapsulated by the hydrogel network. Therefore, when administered in vivo, semaglutide may be rapidly released within a short period, leading to excessively high C levels that may cause unintended gastrointestinal side effects. max value.

[0132] Therefore, the compositions described herein may comprise at least one dispersant that inhibits the aggregation of therapeutic peptides (e.g., by weakening or otherwise disrupting the interaction between the therapeutic peptide and other therapeutic peptides) to enhance the binding of the therapeutic peptide to the dynamic hydrogel. For example, the dispersant may comprise one or more surfactants, such as nonionic surfactants, anionic surfactants, cationic surfactants, and / or zwitterionic surfactants. Examples of surfactants that may be used include polysorbates (e.g., polysorbate 20 (Tween 20), polysorbate 40 (Tween 40), polysorbate 60 (Tween 60), polysorbate 80 (Tween 80)), and sorbitol fatty acid esters (e.g., sorbitol monolaurate (Span 20), sorbitol monostearate (Span 60), sorbitol tristearate (Span 20), etc.). 65), poloxamer (e.g., P188, P237, P338, P407), polyoxyethylene alkyl ethers (e.g., Brij surfactants), alkyl sulfates (e.g., dodecyl sulfates) and their salts (e.g., sodium dodecyl sulfate (SDS)), fatty acids (e.g., lauric acid, myristic acid, palmitic acid, stearic acid), fatty alcohols (e.g., lauryl alcohol, myristol, palmitol, stearyl alcohol), phospholipids, and other lipids or derivatives thereof that exhibit surfactant behavior. As another example, the dispersant may include one or more tonicity agents, such as alcohol-related tonicity agents (e.g., propylene glycol, glycerol, mannitol). Optionally, the compositions herein may comprise a combination of two or more dispersants, such as surfactants and tonicity agents (e.g., Tween 20 and polysorbate). The dispersant may be mixed with the therapeutic peptide prior to combination with the components of the dynamic hydrogel.

[0133] In some embodiments, the concentration of the dispersant is sufficient to inhibit the aggregation of the therapeutic peptide, but low enough to avoid interfering with the interaction between the therapeutic peptide and the dynamic hydrogel, and / or to avoid interfering with hydrogel formation. For example, the concentration of the dispersant in the composition may not exceed 50 mg / mL, 40 mg / mL, 30 mg / mL, 25 mg / mL, 20 mg / mL, 15 mg / mL, 10 mg / mL, 5 mg / mL, 4 mg / mL, 3 mg / mL, 2 mg / mL, 1.5 mg / mL, 1 mg / mL, or 0.5 mg / mL. The concentration of the dispersant in the composition can be 0.5 mg / mL to 50 mg / mL, 0.5 mg / mL to 25 mg / mL, 0.5 mg / mL to 15 mg / mL, 0.5 mg / mL to 10 mg / mL, 0.5 mg / mL to 5 mg / mL, 0.5 mg / mL to 2 mg / mL, 0.5 mg / mL to 1 mg / mL, 1 mg / mL to 50 mg / mL, 1 mg / mL to 25 mg / mL, 1 mg / mL to 15 mg / mL, 1 mg / mL to 10 mg / mL, 1 mg / mL to 5 mg / mL, 1 mg / mL to 2 mg / mL, 2 mg / mL to 50 mg / mL, 2 mg / mL to 25 mg / mL, 2 mg / mL to 15 mg / mL, 2 mg / mL to 10 mg / mL, 2 mg / mL to 5 mg / mL, 5 mg / mL to 50 mg / mL, 5 mg / mL to 25 mg / mL, 5 mg / mL to 15 mg / mL. mg / mL, 5 mg / mL to 10 mg / mL, 10 mg / mL to 50 mg / mL, 10 mg / mL to 25 mg / mL, 10 mg / mL to 15 mg / mL, 15 mg / mL to 50 mg / mL, 15 mg / mL to 25 mg / mL, or 25 mg / mL to 50 mg / mL.

[0134] The concentration of the dispersant in the composition can depend on the type of dispersant used. For example, in embodiments where the dispersant is or contains a surfactant (e.g., Tween 20), the surfactant may be present in the composition at a concentration of less than or equal to 5 mg / mL, 4 mg / mL, 3 mg / mL, 2 mg / mL, 1.5 mg / mL, 1.2 mg / mL, 1.1 mg / mL, 1 mg / mL, or 0.5 mg / mL. The surfactant concentration may be from 0.1 mg / mL to 5 mg / mL, 0.5 mg / mL to 1.5 mg / mL, or 1 mg / mL to 2 mg / mL. As another example, in embodiments where the dispersant is or contains an osmotic pressure regulator (e.g., propylene glycol), the osmotic pressure regulator may be present in the composition at a concentration of 1 mg / mL to 50 mg / mL, 5 mg / mL to 40 mg / mL, 10 mg / mL to 30 mg / mL, or 16 mg / mL to 26 mg / mL.

[0135] In some embodiments, the compositions herein contain little or no saline solution. Saline solution can increase hydrophobic interactions that promote the aggregation of therapeutic peptides. Therefore, removing saline solution from the therapeutic peptide solution reduces aggregation and enhances the binding of the therapeutic peptides to the dynamic hydrogel. In some embodiments, the concentration of sodium chloride in the composition does not exceed 5 mg / mL, 4 mg / mL, 3 mg / mL, 2 mg / mL, 1.5 mg / mL, 1 mg / mL, 0.5 mg / mL, 0.25 mg / mL, 0.1 mg / mL, 0.05 mg / mL, or 0.01 mg / mL. In some embodiments, the compositions herein do not contain any sodium chloride.

[0136] However, in other embodiments, the size of the therapeutic peptide may be larger than the pore size of the dynamic hydrogel, for example if the therapeutic peptide is conjugated, bound, or otherwise linked to a larger molecule or component (e.g., a protein or microparticle), and / or if the therapeutic peptide precipitates under formulation conditions to form larger aggregates. For example, abiglutide and dulaglutide are conjugated to proteins or protein fragments, and exenatide-LAR is encapsulated in biodegradable microparticles. In such embodiments, the therapeutic peptide can be encapsulated in the dynamic hydrogel via physical embedding of the hydrogel network.

[0137] In embodiments where the therapeutic peptide comprises a lipophilic substituent (e.g., a fatty acid side chain), the composition may further comprise a binding agent that binds to the lipophilic substituent and / or otherwise interacts with the lipophilic substituent (e.g., via non-covalent interactions such as hydrophobic interactions). For example, the binding agent may be albumin. The binding agent may be large enough to be physically embedded within the hydrogel network of a dynamic hydrogel. Thus, when a larger molecule is present in the composition, the therapeutic peptide can bind to the binding agent via the interaction between the lipophilic substituent and the binding agent, and is therefore physically embedded within the dynamic hydrogel together with the binding agent. In some embodiments, the binding agent is present in the composition in an amount of 0.1 molar equivalent to 1 molar equivalent relative to the amount of therapeutic peptide in the composition. In some embodiments, at least 1 molar equivalent of the therapeutic peptide is present at each binding site on the binding agent (e.g., albumin has binding sites for two lipophilic substituents). The binding agent may be used in combination with or without a dispersant.

[0138] The compositions described herein can be administered to the subject via any suitable route (e.g., parenteral route). For example, in some embodiments, the compositions are administered to the subject via injection (e.g., subcutaneous or intramuscular injection). The shear-thinning properties of the dynamic hydrogel allow for delivery via injection, while the self-healing properties of the dynamic hydrogel allow for the formation of a reservoir at the injection site, which generates a controlled release of therapeutic peptides during the desired therapeutic period. Injection of the compositions can be performed using any suitable tubular device having a lumen configured for delivery of the hydrogel, such as needles (e.g., hypodermic needles, surgical needles, infusion needles), syringe pens, catheters, cannulas, tubing, etc. The compositions can be injected into any suitable site of the subject's body, such as the arm, thigh, abdomen, or buttock. The compositions can be formulated to have a sufficiently small volume for injection, such as less than or equal to 2 mL, 1.75 mL, 1.5 mL, 1.25 mL, 1 mL, 0.75 mL, 0.5 mL, or 0.25 mL. In some embodiments, the composition is administered as a single injection at a single injection site, while in other embodiments, the composition may be administered as multiple injections at the same or different injection sites. The composition may be administered to the subject at any suitable frequency, such as once a week, once every two weeks, once every four weeks, once a month, once every two months, once every three months, once every four months, once every five months, once every six months, once every nine months, or once a year.

[0139] The compositions described herein can be configured to deliver a therapeutically effective amount of a therapeutic peptide during a desired therapeutic period, said therapeutic peptide being an amount that effectively improves or prevents symptoms of a disease or condition in a subject. For example, the therapeutic period can be at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 28 days, 35 days, 40 days, 50 days, 60 days, 70 days, 80 days, 90 days, 100 days, 110 days, 120 days, 150 days, 180 days, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or 1 year. The therapeutic period can also be approximately 2 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 9 months, or 12 months. As described elsewhere herein, the therapeutic period can be adjusted by regulating the degradation rate of the hydrogel.

[0140] During the treatment period, the composition can deliver the therapeutic peptide at a rate of approximately 0.1 mg / week, 0.25 mg / week, 0.5 mg / week, 1 mg / week, 1.5 mg / week, 2 mg / week, 2.5 mg / week, 3 mg / week, 4 mg / week, 5 mg / week, 6 mg / week, 7 mg / week, 8 mg / week, 9 mg / week, 10 mg / week, 11 mg / week, 12 mg / week, 15 mg / week, 20 mg / week, or 25 mg / week. The delivery rate can be 0.1 mg / week to 25 mg / week, 0.25 mg / week to 25 mg / week, 0.5 mg / week to 20 mg / week, 0.5 mg / week to 2 mg / week, 0.5 mg / week to 1.5 mg / week, 1 mg / week to 2 mg / week, 2 mg / week to 5 mg / week, 5 mg / week to 15 mg / week, 5 mg / week to 10 mg / week, 10 mg / week to 20 mg / week, 10 mg / week to 15 mg / week, 12 mg / week to 25 mg / week, 12 mg / week to 15 mg / week, 15 mg / week to 25 mg / week, 15 mg / week to 20 mg / week, or 20 mg / week to 25 mg / week. Optionally or in combination, the composition can deliver the therapeutic peptide at rates of approximately 0.5 wt% / day, 0.6 wt% / day, 0.7 wt% / day, 0.8 wt% / day, 0.9 wt% / day, 1 wt% / day, 1.25 wt% / day, 1.5 wt% / day, 1.75 wt% / day, 2 wt% / day, 2.5 wt% / day, 3 wt% / day, 4 wt% / day, or 5 wt% / day (the wt% of GLP-1 RA can be measured relative to the total amount of therapeutic peptide initially present in the composition).

[0141] In some embodiments, when administered to a subject in vivo, the composition produces a steady-state concentration (C0) of the therapeutic peptide in serum of approximately 10 ng / mL, 25 ng / mL, 50 ng / mL, 75 ng / mL, 100 ng / mL, 150 ng / mL, 200 ng / mL, 250 ng / mL, 300 ng / mL, 350 ng / mL, 400 ng / mL, 500 ng / mL, 600 ng / mL, 700 ng / mL, 800 ng / mL, 900 ng / mL, 1000 ng / mL, 1500 ng / mL, or 2000 ng / mL. 稳态 ) and / or steady-state average concentration. C of the therapeutic peptide in serum produced by the composition. 稳态 Can be 10 ng / mL to 2000 ng / mL, 10 ng / mL to 1000 ng / mL, 10 ng / mL to 500 ng / mL, 10 ng / mL to 100 ng / ml, 10 ng / mL to 50 ng / mL, 10 ng / mL to 25ng / mL, 50 ng / mL to 2000 ng / mL, 50 ng / mL to 1000 ng / mL, 50 ng / mL to 500 ng / mL, 50 ng / mL to 100 ng / mL, 100 ng / mL to 2000 ng / mL, 100 ng / mL to 1000 ng / mL, 100 ng / mL to 500 ng / mL, 100 ng / mL to 300 ng / mL, 150 ng / mL to 250 ng / mL, 500 ng / mL to 2000 ng / mL, 500 ng / mL to 1000 ng / mL, or 1000 ng / mL to 2000 ng / mL. C of therapeutic peptides. 稳态 This can be achieved on day 1, day 2, day 3, day 4, day 5, day 6, or day 7 after administration of the composition to the subject. In some embodiments, when administered to the subject in vivo, the composition produces a maximum concentration (C0) of the therapeutic peptide in serum less than or equal to 2000 ng / mL, 1500 ng / mL, 1000 ng / mL, or 500 ng / mL. max In some embodiments, when administered to a subject in vivo, the composition produces a therapeutic peptide in serum with a C-value of... max The C-value of therapeutic peptides in serum does not exceed [a certain percentage]. 稳态 And / or 1000 times, 500 times, 200 times, 100 times, 50 times or 10 times the steady-state average concentration.

[0142] In some embodiments, this technology provides a method of treating a subject by applying a composition as described herein. The composition can treat a subject's disease or condition by producing a desired therapeutic effect in the subject, such as alleviating symptoms, reducing the severity of the disease or condition, inhibiting the underlying cause of the disease or condition, stabilizing the disease or condition in a non-late stage, delaying the progression of the disease or condition, and / or improving or alleviating the disease or condition. Examples of diseases and conditions that can be treated with the compositions described herein include diabetes and / or related conditions (e.g., prediabetes, type 1 diabetes, type 2 diabetes, hyperglycemia, impaired glucose tolerance), obesity or overweight, eating disorders (e.g., bulimia nervosa, binge eating disorder), obstructive sleep apnea, cardiovascular diseases (e.g., hypertension, atherosclerosis, myocardial infarction, coronary artery disease), liver diseases (e.g., non-alcoholic fatty liver disease), neurological diseases and / or neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, peripheral neuropathy, ischemia, stroke, multiple sclerosis), inflammatory diseases (e.g., asthma, psoriasis, inflammatory bowel disease), kidney diseases, bone diseases (e.g., bone fragility, osteoporosis), hormonal disorders (e.g., polycystic ovary syndrome), and gastrointestinal diseases (e.g., short bowel syndrome).

[0143] In some embodiments, methods of treating diabetes and / or diabetes-related conditions (e.g., prediabetes, type 1 diabetes, type 2 diabetes, hyperglycemia, impaired glucose tolerance) include administering a composition of the present technology to a subject in need. The composition may be administered to the subject via a single injection (e.g., subcutaneous or intramuscular injection). The composition may comprise a dynamic hydrogel (e.g., PNP hydrogel) encapsulating a therapeutically effective amount of a therapeutic peptide (e.g., an incretin mimic, such as GLP-1 RA) for the treatment of diabetes and / or diabetes-related conditions. For example, a therapeutically effective amount may be the amount of a therapeutic peptide that results in a sustained reduction and / or regulation of the subject's blood glucose levels during treatment. The effective therapeutic dose can be 0.1 mg / week to 25 mg / week, 0.1 mg / week to 5 mg / week, 0.5 mg / week to 20 mg / week, 0.5 mg / week to 2 mg / week, 0.75 mg / week to 1.5 mg / week, 1 mg / week to 2 mg / week, 5 mg / week to 10 mg / week, 10 mg / week to 20 mg / week, or 10 mg / week to 15 mg / week. The effective therapeutic dose can vary depending on the type of therapeutic peptide. For example, for semaglutide, the effective therapeutic dose may be approximately 1 mg / week; for liraglutide, it may be approximately 12.6 mg / week (1.8 mg / day); for telpoglycinide, it may be approximately 2.5 mg / week, 5 mg / week, 7.5 mg / week, 10 mg / week, 12.5 mg / week, or 15 mg / week; and for retaglutide, it may be approximately 1 mg / week, 4 mg / week, 8 mg / week, or 12 mg / week. The components of the dynamic hydrogel can be selected to provide injectability, in vivo cohesive reservoir formation, and controlled release of the therapeutic peptide for a sufficiently long period to treat diabetes and / or diabetes-related conditions. For example, the treatment period may be at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 9 months, or 12 months. For example, dynamic hydrogels can be PNP hydrogels (e.g., composed of hydrophobically modified cellulose derivatives (e.g., HPMC-C12) and multiple amphiphilic nanoparticles (e.g., PEG-PLA nanoparticles)).PNP hydrogels can be 0.5-5 hydrogel, 0.5-8 hydrogel, 0.5-10 hydrogel, 0.5-12 hydrogel, 0.5-15 hydrogel, 0.8-5 hydrogel, 0.8-8 hydrogel, 0.8-10 hydrogel, 0.8-12 hydrogel, 0.8-15 hydrogel, 1-5 hydrogel, 1-8 hydrogel, 1-10 hydrogel, 1-12 hydrogel, 1-15 hydrogel, 1.5-5 hydrogel, 1.5-8 hydrogel, 1.5-10 hydrogel, 1.5-12 hydrogel, 1.5-15 hydrogel, 2-5 hydrogel, 2-8 hydrogel, 2-10 hydrogel, 2-12 hydrogel, 2-15 hydrogel, 3-5 hydrogel, 3-8 hydrogel, 3-10 hydrogel, 3-12 hydrogel, or 3-15 hydrogel. Optionally, the dynamic hydrogel may contain at least one dispersant that inhibits the aggregation of therapeutic peptides, such as a surfactant (e.g., Tween 20) and / or an osmotic pressure regulator (e.g., propylene glycol).

[0144] In some embodiments, methods of treating obesity and / or weight loss include administering the composition of this technology to a subject in need. The composition may be administered to the subject via a single injection (e.g., subcutaneous or intramuscular). The composition may comprise a dynamic hydrogel (e.g., PNP hydrogel) encapsulating a therapeutically effective amount of a therapeutic peptide (e.g., an incretin mimic, such as GLP-1 RA) for treating the subject's obesity and / or reducing the subject's weight. For example, a therapeutically effective amount may be the amount of a therapeutic peptide that results in a desired amount of weight loss and / or maintenance of weight within a desired range. The therapeutically effective amount may be 0.1 mg / week to 25 mg / week, 0.1 mg / week to 5 mg / week, 0.5 mg / week to 20 mg / week, 0.5 mg / week to 2 mg / week, 0.75 mg / week to 1.5 mg / week, 1 mg / week to 2 mg / week, 5 mg / week to 10 mg / week, 10 mg / week to 20 mg / week, or 10 mg / week to 15 mg / week. The effective therapeutic dose can vary depending on the type of therapeutic peptide. For example, for semaglutide, the effective therapeutic dose may be approximately 1 mg / week; for liraglutide, it may be approximately 12.6 mg / week (1.8 mg / day); for telpoglutide, it may be approximately 2.5 mg / week, 5 mg / week, 7.5 mg / week, 10 mg / week, 12.5 mg / week, or 15 mg / week; and for retaglutide, it may be approximately 1 mg / week, 4 mg / week, 8 mg / week, or 12 mg / week. The components of the dynamic hydrogel can be selected to provide injectability, in vivo cohesive reservoir formation, and controlled release of the therapeutic peptide over a sufficiently long period for the treatment of obesity and / or weight loss. For example, the treatment period may be at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 9 months, or 12 months. For example, dynamic hydrogels can be PNP hydrogels (e.g., composed of hydrophobically modified cellulose derivatives (e.g., HPMC-C12) and multiple amphiphilic nanoparticles (e.g., PEG-PLA nanoparticles)). PNP hydrogels can be 0.5-5 hydrogel, 0.5-8 hydrogel, 0.5-10 hydrogel, 0.5-12 hydrogel, 0.5-15 hydrogel, 0.8-5 hydrogel, 0.8-8 hydrogel, 0.8-10 hydrogel, 0.8-12 hydrogel, 0.8-15 hydrogel, 1-5 hydrogel, 1-8 hydrogel, 1-10 hydrogel, 1-12 hydrogel, 1-15 hydrogel, 1.5-5 hydrogel, 1.5-8 hydrogel, 1.5-10 hydrogel, 1.5-12 hydrogel, 1.5-15 hydrogel, 2-5 hydrogel, 2-8 hydrogel, 2-10 hydrogel, 2-12 hydrogel, 2-15 hydrogel, 3-5 hydrogel, 3-8 hydrogel, 3-10 hydrogel, 3-12 hydrogel, or 3-15 hydrogel.Optionally, the dynamic hydrogel may contain at least one dispersant that inhibits the aggregation of therapeutic peptides, such as a surfactant (e.g., Tween 20) and / or an osmotic pressure regulator (e.g., propylene glycol).

[0145] In some embodiments, this technology provides a method for preparing a composition for treating a disease or condition as described herein. The method may include combining components of a dynamic hydrogel (e.g., polymers and nanoparticles) with a therapeutic peptide to form a dynamic hydrogel encapsulating the therapeutic peptide. The combination of the hydrogel components and the therapeutic peptide may be performed using simple mixing at mild conditions (e.g., physiological pH (e.g., pH 7.0 to 7.4)) at room temperature (e.g., 25°C) or at a physiological temperature (e.g., 37°C). Optionally, the method may include combining the hydrogel components and the therapeutic peptide with other components (e.g., dispersants and / or additional therapeutic agents (e.g., antidiabetic agents or anti-obesity agents)). In some embodiments, the composition is prepared no more than 1 hour, 30 minutes, 15 minutes, 10 minutes, 5 minutes, 2 minutes, or 1 minute prior to application to a subject. Optionally or in combination, the composition may be prepared at least 1 minute, 2 minutes, 5 minutes, 10 minutes, 15 minutes, 30 minutes, or 1 hour prior to application to a subject. A majority or all of the dynamic hydrogel may be formed prior to application to a subject. For example, dynamic hydrogels can be sufficiently cross-linked (e.g., non-covalently cross-linked) to exhibit the shear-thinning, self-healing, and / or viscoelastic properties described herein prior to application of the composition to a subject.

[0146] In some embodiments, this technology provides kit products for preparing compositions as described herein. The kit product may comprise a solution containing a therapeutic peptide and one or more solutions containing a dynamic hydrogel component (e.g., a solution containing a polymer and a solution containing nanoparticles, or a single solution containing both polymers and nanoparticles). The therapeutic peptide solution may contain a dispersant (e.g., a surfactant and / or an osmotic regulator). Optionally, the kit product may contain a solution containing an additional therapeutic agent. The solutions may be provided in tubes, bottles, ampoules, syringes, or any other suitable storage container. In some embodiments, each solution independently contains a suitable pharmaceutically acceptable diluent. A pharmaceutically acceptable diluent can be any diluent that does not substantially produce adverse reactions (e.g., toxicity, allergic reactions, or immune responses) when administered to a subject. Examples of pharmaceutically acceptable diluents include, but are not limited to, saline, Ringer's solution, glucose solution, phosphate-buffered saline, water, or combinations thereof. Pharmaceutically acceptable diluents may include isotonic imparting agents such as sodium chloride, potassium chloride, or monosodium phosphate. Pharmaceutically acceptable diluents may include buffers such as bicarbonate, TRIS, HEPES, MOPS, CHES, CHAPS, or phosphate-buffered saline. Pharmaceutically acceptable diluents may appropriately include stabilizers and / or preservatives. Additional examples and details of pharmaceutically acceptable diluents can be found in Martin, Remington's Pharmaceutical Sciences, 21st Ed., Mack Publ. Co., Easton, Pa. (2005), which is incorporated herein by reference in its entirety.

[0147] Example

[0148] This technology is further illustrated by the following non-limiting embodiments.

[0149] Example 1: Preparation of PNP hydrogel loaded with incretin mimicry

[0150] This embodiment describes a method for preparing a PNP hydrogel loaded with an incretin mimic. In short, a clinically used incretin mimic is dissolved in a buffer solution and mixed with polymer nanoparticles, subsequently mixed with a hydrophobically modified cellulose polymer to form a shear-thinning, self-healing hydrogel. Different incretin mimics can be used in the hydrogel, and additives can be included to stabilize the cargo and / or adjust release properties.

[0151] Smegglutide, liraglutide, and telposide are GLP-1RAs carrying a single optimized fatty acid and linker modification, which allows reversible binding to albumin to prolong the circulating half-life while maintaining optimal potency. While reducing the treatment frequency from daily to weekly is associated with improved patient adherence, there is still room for improvement in reducing treatment burden and enhancing patient adherence. Figure 2A To address this challenge, long-acting formulations of smegglutide and telpolide have been developed to provide continuous treatment for more than four months from a single dose, consistent with the typical cycle of type 2 diabetes patients visiting their endocrinologist or primary care provider. Figure 2B ).

[0152] HPMC (compliant with USP testing specifications), N,N-diisopropylethylamine (Hunig base), hexane, diethyl ether, N-methyl-2-pyrrolidone (NMP), dichloromethane (DCM), lactide (LA), 1-dodecylisocynate, and diazobicylcoundecene (DBU) were purchased from Sigma-Aldrich and used as is. Monomethoxy-PEG (5 kDa) was purchased from Sigma-Aldrich and dried under vacuum before use. Glassware and stir bar were dried in an oven at 180°C. When specified, the solvent was degassed by three cycles of freezing, pumping, and thawing.

[0153] Dodecyl-modified (hydroxypropyl)methylcellulose (HPMC-C12) was prepared as follows: HPMC (1.0 g) was dissolved in NMP (40 mL) by stirring at 80 °C for 1 h. When the solution reached room temperature (RT), 1-dodecyl isocyanate (105 mg, 0.5 mmol) and N,N-diisopropylethylamine (catalyst, about 3 drops) were dissolved in NMP (5.0 mL). This solution was added dropwise to the reaction mixture, and the mixture was stirred at room temperature for 16 h. The solution was then precipitated from acetone, decanted, redissolved in water (about 2 wt%), and dialyzed in a dialysis tube for 3–4 days. The polymer was lyophilized and reconstituted into a 60 mg / mL solution using sterile PBS.

[0154] PEG-PLA nanoparticles (PEG-PLA NP) were prepared as follows: Monomethoxy-PEG (5 kDa; 0.25 g, 4.1 mmol) and DBU (15 µL, 0.1 mmol; relative to LA 1.4 mol%) were dissolved in anhydrous dichloromethane (1.0 mL). LA (1.0 g, 6.9 mmol) was dissolved in anhydrous DCM (3.0 mL) with gentle heating. The LA solution was rapidly added to the PEG / DBU solution and stirred for 10 min. The reaction mixture was quenched and precipitated by a 1:1 hexane and diethyl ether solution. The synthesized PEG-PLA was collected and dried under vacuum. The molecular weight and dispersibility of the polymer were verified to meet the quality control (QC) parameters using gel permeation chromatography (GPC). Figure 3A A 1 mL solution of PEG-PLA in DMSO (50 mg / mL) was added dropwise to 10 mL of water at room temperature with high stirring (600 rpm). The nanoparticles were purified by centrifugation through a filter (10 kDa molecular weight cutoff; Millipore Amicon Ultra-15) and then resuspended in PBS to a final concentration of 200 mg / mL. The nanoparticles were characterized by dynamic light scattering (DLS) to determine the nanoparticle diameter (35 ± 4 nm). Figure 3B ).

[0155] The hydrogel formulation contains 1 wt% HPMC-C12 and 10 wt% PEG-PLA nanoparticles and is designated PNP-1-10. These hydrogels are prepared by mixing a 6 wt% HPMC-C12 polymer solution, a 20 wt% nanoparticle solution, and PBS or water containing GLP-1 RA (semaglutide, liraglutide, or telpolide). Semaglutide, liraglutide, and telpolide are obtained from the Stanford University Hospital Formulary as pharmaceutical products Ozempic® (Novo Nordisk), Victoza® (Eli Lilly), and Mountaro® (Lilly), respectively. These pharmaceutical products are provided as aqueous formulations comprising phosphate buffer, osmolarity regulators (e.g., propylene glycol (Ozempic® and Victoza®) or saline (Mounjaro®)), and preservatives (e.g., phenol). These formulations were lyophilized and then resuspended in water to prepare hydrogels. For formulations containing Tween 20 and / or α-cyclodextrin (aCD), these additives were mixed with GLP-1 RA, and the resulting solution was mixed with nanoparticles. The nanoparticles and aqueous components were loaded into one syringe, HPMC-C12 was loaded into a second syringe, and the components were mixed using a bent connector. Figure 4 After mixing, replace the bent tip with a 21-gauge needle for injection. Smegglutide, liraglutide, and telpopotide hydrogel formulations and their corresponding components are listed in Tables 2A and 2B, 3A and 3B, and 4A and 4B, respectively.

[0156] Table 2A: Smegglutinin PNP hydrogel formulation

[0157] Table 2B: Smegglutinin PNP hydrogel formulation

[0158] Table 3A: Liraglutide PNP hydrogel formulation

[0159] Table 3B: Liraglutide PNP hydrogel formulation

[0160] Table 4A: PNP hydrogel formulations for telpopeptide

[0161] Table 4B: PNP hydrogel formulations for telpopeptide

[0162] Example 2: Rheological characterization of PNP hydrogels loaded with incretin mimics

[0163] This embodiment describes the rheological characterization of a PNP hydrogel loaded with an incretin mimic. Rheological tests were performed on a stress-controlled TA Instruments DHR-2 rheometer using a 20 mm diameter serrated parallel plate with a 600 μm gap. All experiments were conducted at 25 °C. Frequency scans were performed from 0.1 rad / s to 100 rad / s at constant oscillatory strain (1%) within the linear viscoelastic region. Amplitude scans were performed at a constant angular frequency of 10 rad / s with strains from 0.01% to 10000% and a gap height of 500 µm. Steady-state sensing was performed from low stress to high stress. Steady-state shear experiments were conducted at low shear rates (0.1 s⁻¹). -1 ) and high shear rate (10 s) -1 The shear rate scan was performed alternately between 10 s and 60 s, for three complete cycles. -1 up to 0.001 s -1 The stress-controlled yield stress measurement (stress scan) is performed using steady-state sensing from low stress to high stress, at 10 points per decimal.

[0164] Figure 5A and Figure 5B This is a graph showing the rheological characterization of the PNP-1-10 hydrogel formulation with varying semaglutide concentrations: frequency-dependent oscillatory shear scan ( Figure 5A ) and stress-dependent oscillatory shear scan ( Figure 5B ).

[0165] Figure 6A and Figure 6B This is a graph showing the rheological characterization of the PNP-2-10 hydrogel formulation with varying semaglutide concentrations: frequency-dependent oscillatory shear scan ( Figure 6A ) and stress-dependent oscillatory shear scan ( Figure 6B ).

[0166] Figures 7A-7C This is a graph showing the rheological characterization of the PNP-1-10 hydrogel formulation with varying telpoide concentrations: frequency-dependent oscillatory shear scan ( Figure 7A and Figure 7C ) and stress-dependent oscillatory shear scan ( Figure 7B ).exist Figure 7CIn this context, PNP-TZP-4.5 mg 1-10 corresponds to PNP-1-10-T9, PNP-TZP-0.28 mg 1-10 corresponds to PNP-1-10-T5, PNP-TZP-4.5 mg 0.1 wt% Tween 1-10 corresponds to PNP-1-10-T11, and PNP-TZP-4.5 mg 0.1 wt% Tween 1-10 corresponds to PNP-1-10-T12.

[0167] These results indicate that the addition of GLP-1 RA and other formulation excipients does not affect the mechanical behavior of the hydrogel.

[0168] Example 3: In vitro release kinetics of smegglutinin from PNP hydrogel formulation

[0169] This example describes the in vitro characterization of semaglutide release from a PNP hydrogel formulation. In vitro release assays were used to investigate the release behavior of semaglutide from the PNP hydrogel. Figure 8A 100 µL of each hydrogel formulation was loaded into a four-inch capillary tube, and 400 µL of PBS medium was slowly added to the top. The surrounding PBS was removed for analysis at 1 h, 3 h, 6 h, 12 h, 24 h, and 48 h after capillary injection, and at one week and two weeks later, with fresh PBS replaced after each aliquot. Smegglutinin was quantified by ELISA to determine the release kinetics over time.

[0170] Figure 8B and Figure 8C This shows that over a two-week period, under low, medium, and high semaglutide loading, semaglutide decreased from PNP-2-10 (… Figure 8B ) and PNP-1-10 ( Figure 8C The graph shows the in vitro release curves representing the cumulative release percentage of the hydrogel formulations. For all formulations, a significant proportion of the semaglutide cargo was released within two weeks. These results suggest that semaglutide-loaded hydrogels may contain a significant portion of “free” cargo (not adhered to the PNP hydrogel matrix), which undergoes rapid diffusion-based release. Semaglutide can form robust dimers at formulation-related concentrations, and these dimers are presumed to constitute the “free” portion of the drug cargo. The semaglutide dimers are small enough (RH < 2 nm) that they are significantly smaller than the relatively large pore size of the PNP hydrogel. It is released from the PNP hydrogel in a relatively short time range (approximately 3.5 nm).

[0171] Figure 8DThis is a graph showing the in vitro release curves of the cumulative release percentage of semaglutide from various PNP-1-10 hydrogel formulations over a two-week period, illustrating the effect of Tween 20 on semaglutide release in the presence of propylene glycol and saline. Over the two-week period, all formulations exhibited low burst release and high cargo retention, primarily attributed to the presence of propylene glycol. The release rate further decreased with increasing Tween 20 levels. It is hypothesized that propylene glycol and Tween 20 act as dispersants, inhibiting the formation of semaglutide dimers and thus promoting stronger binding of the fatty acid side chains of semaglutide to the PNP hydrogel matrix.

[0172] Figure 8E This graph shows the in vitro release curves of semaglutide from various PNP-1-10 hydrogel formulations over a two-week period, demonstrating that the addition of α-cyclodextrin (aCD) had a negligible effect on semaglutide release in the presence of propylene glycol and saline. All formulations exhibited low burst release and high cargo retention over the two-week period.

[0173] Figure 8F This is a graph showing the in vitro release curves of the cumulative release % of semaglutide from various PNP-1-10 hydrogel formulations over a two-week period, illustrating the effect of adding Tween 20 on semaglutide release in the presence and absence of saline. Over the two-week period, all formulations exhibited low burst release and high cargo retention, primarily attributed to the presence of propylene glycol. The release rate further decreased with increasing Tween 20 content. The formulation containing Tween 20 in the absence of saline showed a modest improvement in cargo retention compared to the formulation containing Tween 20 in the presence of saline. Figure 8D The presence of saline solution can increase the hydrophobic interactions that promote the dimerization of smegglutinin. Therefore, removing the saline solution before formulation into a hydrogel can promote the depolymerization of smegglutinin dimers and improve the binding of smegglutinin to the hydrogel.

[0174] Figure 8GThis is a graph showing the in vitro release curves of semaglutide from various PNP-1-10 hydrogel formulations, illustrating the cumulative release percentage, and demonstrating the role of bovine serum albumin (BSA) in the release buffer and hydrogel. The PNP-1-10-S7 hydrogel formulation was prepared with or without 1% BSA in the hydrogel and in the PBS release buffer. The PNP-1-10-S10 hydrogel formulation was prepared with 3.59 mg / mL BSA in the hydrogel and without BSA in the release buffer. The PNP-1-10-S16 formulation was prepared without BSA in either the release buffer or the hydrogel. No significant differences in release rates were observed between the groups, thus indicating that the presence of BSA around the hydrogel does not affect the release of semaglutide.

[0175] Example 4: In vitro release kinetics of liraglutide from PNP hydrogel formulation

[0176] This example describes the in vitro characterization of liraglutide release from a PNP hydrogel formulation. The in vitro release assay of Example 3 was used to investigate the release behavior of liraglutide from the PNP hydrogel.

[0177] Figure 9 This is a graph showing the in vitro release curves of the cumulative release percentage of liraglutide from 1.8 mg / mL PNP-1-10 and PNP-2-10 hydrogel formulations over a two-week period. Even in the absence of propylene glycol and Tween 20, the formulations still exhibited negligible and high cargo retention over the two-week period. It is assumed that substantially all of the liraglutide is "bound" cargo, which adheres to the PNP hydrogel structure and is released primarily through hydrogel erosion, which is strictly limited in this capillary release model. The fatty acid side chains of liraglutide can drive the formation of a more disordered heptamer structure at micromolar concentrations, thus allowing liraglutide to bind more strongly to the structural motifs within the PNP hydrogel, even in the absence of a dispersant.

[0178] Example 5: In vitro release kinetics of telpolide from PNP hydrogel formulation

[0179] This example describes the in vitro characterization of telpolide release from a PNP hydrogel formulation. The release behavior of telpolide from the PNP hydrogel was studied using the in vitro release assay of Example 3. Over a two-week period, the formulation exhibited low levels of burst release and high cargo retention.

[0180] Example 6: In vitro release kinetics of PNP hydrogels formulated with liraglutide and insulin glargine

[0181] This example describes the in vitro characterization of liraglutide and insulin glargine released from a PNP hydrogel formulation.

[0182] Insulin glargine, as the pharmaceutical product Lantus® (Sanofi), was obtained from the Stanford University Hospital Formulary. PNP-1-10 hydrogels containing and without liraglutide and / or insulin glargine were prepared according to the protocol of Example 1. The release behavior of liraglutide and insulin glargine from the PNP hydrogels was studied using the in vitro release assay of Example 3. The hydrogel formulations and their corresponding components are listed in Tables 5A and 5B below.

[0183] Table 5A: Liraglutide and Lantus PNP hydrogel formulations

[0184] Table 5B: Liraglutide and Lantus PNP hydrogel formulations

[0185] Figure 10 This is a graph showing the in vitro release curves of liraglutide and lantus from the PNP-1-10 hydrogel formulation, measured by ELISA, over a two-week period. All formulations exhibited low burst release and high cargo retention over the two-week period, indicating that both liraglutide and insulin glargine can be successfully encapsulated in the PNP hydrogel.

[0186] Example 7: In vivo characterization of PNP hydrogel formulation loaded with incretin mimics

[0187] This embodiment describes the in vivo pharmacokinetics, pharmacodynamics, and biocompatibility characterization of a PNP hydrogel formulation loaded with an incretin mimic in diabetic rats.

[0188] Male Sprague Dawley rats (8-10 weeks old, Charles River) weighing 160-230 g were fasted for 6-8 hours in the morning before treatment with nicotinamide (NA) and streptozotocin (STZ). NA was dissolved in 1X PBS and administered intraperitoneally at a dose of 110 mg / kg. STZ was immediately diluted with sodium citrate buffer to 10 mg / mL before injection. STZ solution was administered intraperitoneally at a dose of 65 mg / kg to each rat. Following STZ injection, rats were provided with water containing 10% sucrose for 24 hours. Blood glucose (BG) levels were measured daily via tail vein sampling using a Bayer Contour Next handheld glucometer to detect hyperglycemia. Type 2 diabetes (T2D) was defined as three consecutive BG measurements of 130-200 mg / dL in non-fasting rats.

[0189] Diabetic rats received either a) a single subcutaneous injection of a PNP hydrogel loaded with semaglutide (1.85 mg / mL) (PNP-1-10-S7 in Tables 2A and 2B) or a PNP hydrogel loaded with telpolide (4.5 mg / mL) (PNP-1-10-T11 in Tables 4A and 4B); or b) daily subcutaneous bolus injections of PBS, 20 µg semaglutide, or 50 µg telpolide. For each treatment group, baseline blood was collected from the tail vein on day zero, and daily blood glucose measurements were collected from the tail vein for 42 days post-treatment using a Bayer Contour Next handheld blood glucose monitor. Blood glucose was measured, and blood samples were immediately collected from the tail vein. Serum semaglutide or telpolide concentrations were measured daily by ELISA for the first seven days of the study, and twice weekly thereafter. Plasma GLP-1 RA concentrations were measured by ELISA at each time point, and the total bioavailability of semaglutide or telpolide was determined at the end of the study. The oral glucose tolerance test was used to group rats according to their glucose tolerance before treatment (on day -1).

[0190] Figure 11 This is a schematic diagram of the treatment plan and timing for blood glucose measurement and serum collection used in the analysis. Diabetic rats received a single subcutaneous injection of PNP hydrogel loaded with semaglutide or telpolide, or daily subcutaneous injections of PBS, 20 µg semaglutide, or 50 µg telpolide daily.

[0191] Figure 12A and Figure 12B This describes the pre-treatment ( Figure 12A ) and 6 weeks after treatment ( Figure 12B A graph showing the results of the oral glucose tolerance test. An oral glucose tolerance test (OGTT) was performed to classify diabetic rats into treatment groups. Rats were fasted before administration of a glucose load via oral gavage. Baseline (fasting) blood glucose was measured before glucose administration, and then at regular intervals. Blood glucose was measured at -5 min, 0 min, 5 min, 15 min, 30 min, 45 min, 60 min, and 120 min. Rats with similar glucose tolerance were paired using the area under the curve (AUC) and then randomly assigned to treatment groups.

[0192] Figure 13 This is a graph showing the percentage change in blood glucose (BG) levels in rats treated with PNP hydrogel relative to those treated with bolus injection. A single administration of semaglutide PNP or telposide PNP hydrogel reduced BG in type 2 diabetes-like male rats over a 6-week period compared to daily PBS bolus injections. This graph shows the change in BG over 6 weeks after each treatment regimen (n = 6).

[0193] Figure 14 This is a graph showing the percentage change in body weight in rats treated with PNP hydrogel relative to those treated with bolus injection. A single administration of semaglutide PNP or telpolide PNP hydrogel reduced the total body weight gain in type 2 diabetes-like male rats over the 6-week treatment period compared to daily PBS bolus injections. This graph shows the change in body weight over 6 weeks in each treatment group (n = 6).

[0194] Figure 15 This figure shows the pharmacokinetics of semaglutide administered daily as a 20 µg bolus injection relative to PNP hydrogel (PNP-1-10 with a 1.8 mg / mL semaglutide loading and 0.05 wt% Tween 20) in male diabetic rats (n = 6) over a 6-week period following treatment.

[0195] Figure 16 This is a graph showing the pharmacokinetics of 20 µg daily bolus injection of semaglutide relative to PNP hydrogel (PNP-1-10 with 1.8 mg / mL semaglutide loading and 0.05 wt% Tween 20) over the first 48 hours in male diabetic rats (n = 6), superimposed on the 24-hour pharmacokinetics of 20 µg intravenous (IV) and subcutaneous (SC) bolus injections.

[0196] Figure 17 The diagram illustrates the use of blood chemistry to assess treatment biocompatibility to observe negative effects on the liver or kidneys and evaluate the treatment's effect on hemoglobin A1c (HbA1c). Blood was collected before treatment and after treatment (6 weeks later). Hepatotoxicity was assessed by measuring alanine aminotransferase (ALT), aspartate aminotransferase (AST), and bilirubin. Nephrotoxicity was assessed by examining creatinine and blood urea nitrogen (BUN) levels. For both the treatment and control groups, ALT, AST, creatine, and BUN values ​​were within the range of healthy rats (defined as mean ± 2 standard deviations).

[0197] These results indicate that the hydrogel formulation effectively maintained treatment-related concentrations of semaglutide and telposide during the six-week study period. Compared to the PBS bolus injection control, the hydrogel-based treatment resulted in a significant reduction in mean body mass (BG) and body weight during the study. The reductions in mean BG and body weight were comparable to those achieved with daily bolus injections of semaglutide and telposide. The hydrogel-based treatment was well-tolerated, with no observable differences in liver and kidney compared to untreated animals.

[0198] Additional instances

[0199] For convenience, additional examples of various aspects of this technology are described below as numbered clauses (1, 2, 3, etc.). These are provided as examples and do not limit the technology of this subject.

[0200] Clause 1. A composition for treating a disease or condition, said composition comprising: Dynamic hydrogel comprising a polymer and a plurality of nanoparticles, wherein the polymer is non-covalently crosslinked with the plurality of nanoparticles; and Acylated peptides, which are encapsulated by the dynamic hydrogel.

[0201] Clause 2. The composition according to Clause 1, wherein the acylated peptide comprises a lipophilic substituent.

[0202] Clause 3. The composition according to Clause 2, wherein the acylated peptide is encapsulated in the dynamic hydrogel via the interaction between the lipophilic substituent and the dynamic hydrogel.

[0203] Clause 4. The composition according to Clause 3, wherein the plurality of nanoparticles have a hydrophobic surface that interacts with the lipophilic substituents of the dynamic hydrogel.

[0204] Clause 5. The composition according to any one of Clauses 2 to 4, wherein the lipophilic substituent comprises an acyl group of a fatty acid.

[0205] Clause 6. The composition according to Clause 5, wherein the fatty acid is a C4 fatty acid, C6 fatty acid, C8 fatty acid, C10 fatty acid, C12 fatty acid, C14 fatty acid, C15 fatty acid, C16 fatty acid, C17 fatty acid, C18 fatty acid, C20 fatty acid, C22 fatty acid, C24 fatty acid, C26 fatty acid, C28 fatty acid, C30 fatty acid, C32 fatty acid, C34 fatty acid, C36 fatty acid, or C38 fatty acid.

[0206] Clause 7. The composition according to any one of Clauses 2 to 6 further comprises a binding agent that interacts with the lipophilic substituent to physically encapsulate the acylated peptide within the dynamic hydrogel.

[0207] Clause 8. The composition according to Clause 7, wherein the binder comprises albumin.

[0208] Clause 9. The composition according to Clause 7 or 8, wherein the binder is larger than the mesh size of the dynamic hydrogel.

[0209] Clause 10. The composition according to any one of Clauses 1 to 9, wherein the acylated peptide is smaller than the pore size of the dynamic hydrogel.

[0210] Clause 11. The composition according to any one of Clauses 1 to 10 further comprises a dispersant that inhibits the aggregation of the acylated peptide.

[0211] Clause 12. The composition according to Clause 11, wherein the dispersant comprises one or more of a surfactant or an osmotic pressure regulator.

[0212] Clause 13. The composition according to Clause 12, wherein the dispersant comprises the surfactant, and the surfactant comprises one or more of polysorbate, sorbitol fatty acid ester, poloxamer, polyoxyethylene alkyl ether, alkyl sulfate, fatty acid, fatty acid alcohol or phospholipid.

[0213] Clause 14. The composition according to Clause 12 or 13, wherein the dispersant comprises the osmotic pressure regulator, and the osmotic pressure regulator comprises one or more of propylene glycol, glycerol, or mannitol.

[0214] Clause 15. The composition according to any one of Clauses 12 to 14, wherein the concentration of the dispersant in the composition is 0.1 mg / mL to 5 mg / mL, 0.5 mg / mL to 1.5 mg / mL, 1 mg / mL to 2 mg / mL, 1 mg / mL to 50 mg / mL, 5 mg / mL to 40 mg / mL, 10 mg / mL to 30 mg / mL, or 16 mg / mL to 26 mg / mL.

[0215] Clause 16. The composition according to any one of Clauses 1 to 15, wherein the acylated peptide comprises an incretin mimic.

[0216] Clause 17. The composition according to Clause 16, wherein the incretin mimic comprises a glucagon-like peptide-1 receptor agonist (GLP-1 RA).

[0217] Clause 18. The composition according to Clause 17, wherein the GLP-1 RA comprises one or more of liraglutide, smegglutide, enoglutide, cotodextrin, mastodextrin, telpogextrin, or retaglutide.

[0218] Clause 19. The composition according to any one of Clauses 16 to 18, wherein the incretin mimic binds only to the GLP-1 receptor.

[0219] Clause 20. The composition according to any one of Clauses 16 to 18, wherein the incretin mimic binds to a GLP-1 receptor and at least one additional receptor.

[0220] Clause 21. The composition according to Clause 20, wherein the at least one additional receptor comprises one or more of glucagon receptor or gastric inhibitory peptide (GIP) receptor.

[0221] Clause 22. The composition according to any one of Clauses 1 to 15, wherein the acylated peptide comprises an analog of a proglucagon-derived peptide.

[0222] Clause 23. The composition according to any one of Clauses 1 to 15, wherein the acylated peptide comprises an amylin analogue.

[0223] Clause 24. The composition according to any one of Clauses 1 to 15, wherein the acylated peptide comprises an insulin analog.

[0224] Clause 25. The composition according to any one of Clauses 1 to 24, wherein the composition comprises 1 mg to 250 mg of the acylated peptide.

[0225] Clause 26. The composition according to any one of Clauses 1 to 25, wherein the concentration of the acylated peptide in the composition is from 1 mg / mL to 200 mg / mL.

[0226] Clause 27. The composition according to any one of Clauses 1 to 26, wherein said polymer comprises a hydrophobically modified polysaccharide.

[0227] Clause 28. The composition according to Clause 27, wherein the hydrophobically modified polysaccharide comprises a hydrophobically modified cellulose derivative.

[0228] Clause 29. The composition according to Clause 28, wherein the hydrophobically modified cellulose derivative is dodecyl modified hydroxypropyl methylcellulose (HPMC-C12).

[0229] Clause 30. The composition according to any one of Clauses 1 to 29, wherein the plurality of nanoparticles comprises a plurality of polymer nanoparticles.

[0230] Clause 31. The composition according to Clause 30, wherein the plurality of polymer nanoparticles are amphiphilic.

[0231] Clause 32. The composition according to Clause 30 or 31, wherein the plurality of polymer nanoparticles are poly(ethylene glycol)-block-poly(lactic acid) (PEG-PLA) nanoparticles.

[0232] Clause 33. The composition according to any one of Clauses 1 to 32, wherein the concentration of the polymer in the dynamic hydrogel is 0.5 wt% to 5 wt%, 0.5 wt% to 4 wt%, 0.5 wt% to 3 wt%, 0.5 wt% to 2 wt%, 0.5 wt% to 1 wt%, 1 wt% to 5 wt%, 1 wt% to 4 wt%, 1 wt% to 3 wt%, 1 wt% to 2 wt%, 2 wt% to 5 wt%, 2 wt% to 4 wt%, 2 wt% to 3 wt%, 3 wt% to 5 wt%, 3 wt% to 4 wt%, or 4 wt% to 5 wt%.

[0233] Clause 34. The composition according to any one of Clauses 1 to 33, wherein the concentration of nanoparticles in the dynamic hydrogel is 1 wt% to 12 wt%, 1 wt% to 10 wt%, 1 wt% to 8 wt%, 1 wt% to 5 wt%, 1 wt% to 3 wt%, 3 wt% to 12 wt%, 3 wt% to 10 wt%, 3 wt% to 8 wt%, 3 wt% to 5 wt%, 5 wt% to 12 wt%, 5 wt% to 10 wt%, 5 wt% to 8 wt%, 8 wt% to 12 wt%, 8 wt% to 10 wt%, or 10 wt% to 12 wt%.

[0234] Clause 35. The composition according to any one of Clauses 1 to 34, wherein the storage modulus of the dynamic hydrogel encapsulating the acylated peptide is 10 Pa to 1000 Pa or 50 Pa to 500 Pa when measured at 25°C at an angular frequency of 0.1 rad / s to 100 rad / s in the linear viscoelastic region of the dynamic hydrogel.

[0235] Clause 36. The composition according to any one of Clauses 1 to 35, wherein the yield stress of the dynamic hydrogel encapsulating the acylated peptide is from 1 Pa to 500 Pa or from 20 Pa to 200 Pa when measured at 25°C.

[0236] Clause 37. The composition according to any one of Clauses 1 to 36, wherein when at 25°C for 1000 s -1 When measured at a shear rate of 100 mPa-s to 1000 mPa-s, the viscosity of the dynamic hydrogel encapsulating the acylated peptide was 100 mPa-s to 1000 mPa-s.

[0237] Clause 38. The composition according to any one of Clauses 1 to 37, wherein, after administration to a subject, the composition delivers the acylated peptide to the subject for a treatment period of at least 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 28 days, 35 days, 40 days, 50 days, 60 days, 70 days, 80 days, 90 days, 100 days, 110 days, 120 days, 150 days, 180 days, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or 1 year.

[0238] Clause 39. The composition according to any one of Clauses 1 to 38, wherein, after administration to a subject, the composition delivers the acylated peptide to the subject at a rate of 0.25 mg / week to 25 mg / week, 0.5 mg / week to 20 mg / week, 0.5 mg / week to 2 mg / week, 0.5 mg / week to 1.5 mg / week, 1 mg / week to 2 mg / week, 2 mg / week to 5 mg / week, 5 mg / week to 15 mg / week, 5 mg / week to 10 mg / week, 10 mg / week to 20 mg / week, 10 mg / week to 15 mg / week, 12 mg / week to 25 mg / week, 12 mg / week to 15 mg / week, 15 mg / week to 25 mg / week, 15 mg / week to 20 mg / week, or 20 mg / week to 25 mg / week.

[0239] Clause 40. The composition according to any one of Clauses 1 to 39, wherein, after administration to a subject, the composition produces in the serum of the subject an average concentration of the acylated peptide of 50 ng / mL to 500 ng / mL, 100 ng / mL to 300 ng / mL, or 150 ng / mL to 250 ng / mL.

[0240] Clause 41. The composition according to any one of Clauses 1 to 40, wherein, after administration to a subject, the composition produces less than or equal to 2000 ng / mL of C in the serum of the subject. max The acylated peptide.

[0241] Clause 42. The composition according to any one of Clauses 1 to 41, wherein the composition is configured for administration via subcutaneous injection.

[0242] Clause 43. The composition according to any one of Clauses 1 to 42, wherein the disease or condition includes diabetes or diabetes-related conditions.

[0243] Clause 44. The composition according to Clause 43, wherein the diabetes or diabetes-related condition includes prediabetes, type 1 diabetes, type 2 diabetes, hyperglycemia, or impaired glucose tolerance.

[0244] Clause 45. The composition according to any one of Clauses 1 to 44, wherein the disease or condition includes one or more of obesity, overweight, eating disorders or obstructive sleep apnea.

[0245] Clause 46. The composition according to any one of Clauses 1 to 45, wherein the disease or condition includes one or more of the following: cardiovascular disease, liver disease, nervous system disease or neurodegenerative disease, inflammatory disease, kidney disease, bone disease, hormonal disease or gastrointestinal disease.

[0246] Clause 47. A method of treating a disease or condition, the method comprising: applying a composition to a subject, wherein the composition comprises: a dynamic hydrogel comprising a polymer and a plurality of nanoparticles, wherein the polymer is non-covalently crosslinked with the plurality of nanoparticles; and an acylated peptide encapsulated by the dynamic hydrogel.

[0247] Clause 48. The method according to Clause 47, wherein the acylated peptide comprises a lipophilic substituent.

[0248] Clause 49. The method according to Clause 48, wherein the acylated peptide is encapsulated in the dynamic hydrogel via the interaction between the lipophilic substituent and the dynamic hydrogel.

[0249] Clause 50. The method according to Clause 49, wherein the plurality of nanoparticles have a hydrophobic surface that interacts with the lipophilic substituents of the dynamic hydrogel.

[0250] Clause 51. The method according to any one of Clauses 48 to 50, wherein the lipophilic substituent comprises an acyl group of a fatty acid.

[0251] Clause 52. The method according to Clause 51, wherein the fatty acid is a C4 fatty acid, C6 fatty acid, C8 fatty acid, C10 fatty acid, C12 fatty acid, C14 fatty acid, C15 fatty acid, C16 fatty acid, C17 fatty acid, C18 fatty acid, C20 fatty acid, C22 fatty acid, C24 fatty acid, C26 fatty acid, C28 fatty acid, C30 fatty acid, C32 fatty acid, C34 fatty acid, C36 fatty acid, or C38 fatty acid.

[0252] Clause 53. The method according to any one of Clauses 48 to 52, wherein the composition further comprises a binding agent that interacts with the lipophilic substituent to physically encapsulate the acylated peptide in the dynamic hydrogel.

[0253] Clause 54. The method according to Clause 53, wherein the binding agent comprises albumin.

[0254] Clause 55. The composition according to Clause 53 or 54, wherein the binder is larger than the mesh size of the dynamic hydrogel.

[0255] Clause 56. The method according to any one of Clauses 47 to 55, wherein the acylated peptide is smaller than the pore size of the dynamic hydrogel.

[0256] Clause 57. The method according to any one of Clauses 47 to 56, wherein the composition further comprises a dispersant that inhibits the aggregation of the acylated peptide.

[0257] Clause 58. The method according to Clause 57, wherein the dispersant comprises one or more of a surfactant or an osmotic pressure regulator.

[0258] Clause 59. The method according to Clause 58, wherein the dispersant comprises the surfactant, and the surfactant comprises one or more of polysorbate, sorbitol fatty acid ester, poloxamer, polyoxyethylene alkyl ether, alkyl sulfate or its salt, fatty acid, and fatty acid alcohol.

[0259] Clause 60. The method according to Clause 58 or 59, wherein the dispersant comprises the osmotic pressure regulator, and the osmotic pressure regulator comprises one or more of propylene glycol, glycerol, or mannitol.

[0260] Clause 61. The method according to any one of Clauses 58 to 60, wherein the concentration of the dispersant in the composition is 0.1 mg / mL to 5 mg / mL, 0.5 mg / mL to 1.5 mg / mL, 1 mg / mL to 2 mg / mL, 1 mg / mL to 50 mg / mL, 5 mg / mL to 40 mg / mL, 10 mg / mL to 30 mg / mL, or 16 mg / mL to 26 mg / mL.

[0261] Clause 62. The method according to any one of Clauses 47 to 61, wherein the acylated peptide comprises an incretin mimic.

[0262] Clause 63. The method according to Clause 62, wherein the incretin mimic comprises a glucagon-like peptide-1 receptor agonist (GLP-1 RA).

[0263] Clause 64. The method according to Clause 63, wherein the GLP-1 RA comprises one or more of liraglutide, smegglutide, enoglutide, cotodextrin, mastodextrin, telpogextrin, or retaglutide.

[0264] Clause 65. The method according to any one of Clauses 62 to 64, wherein the incretin mimic binds only to the GLP-1 receptor.

[0265] Clause 66. The method according to any one of Clauses 62 to 64, wherein the incretin mimic binds to the GLP-1 receptor and at least one additional receptor.

[0266] Clause 67. The method according to Clause 66, wherein the at least one additional receptor comprises one or more of glucagon receptor or gastric inhibitory peptide (GIP) receptor.

[0267] Clause 68. The method according to any one of Clauses 47 to 67, wherein the acylated peptide comprises an analog of a proglucagon-derived peptide.

[0268] Clause 69. The method according to any one of Clauses 47 to 67, wherein the acylated peptide comprises an amylin analogue.

[0269] Clause 70. The method according to any one of Clauses 47-67 or 69, wherein the acylated peptide comprises an insulin analog.

[0270] Clause 71. The method according to any one of Clauses 47 to 70, wherein the composition comprises 1 mg to 250 mg of the acylated peptide.

[0271] Clause 72. The method according to any one of Clauses 47 to 71, wherein the concentration of the acylated peptide in the composition is from 1 mg / mL to 200 mg / mL.

[0272] Clause 73. The method according to any one of Clauses 47 to 72, wherein said polymer comprises a hydrophobically modified polysaccharide.

[0273] Clause 74. The method according to Clause 73, wherein the hydrophobically modified polysaccharide comprises a hydrophobically modified cellulose derivative.

[0274] Clause 75. The method according to Clause 74, wherein the hydrophobically modified cellulose derivative is dodecyl modified hydroxypropyl methylcellulose (HPMC-C12).

[0275] Clause 76. The method according to any one of Clauses 47 to 75, wherein the plurality of nanoparticles comprises a plurality of polymer nanoparticles.

[0276] Clause 77. The method according to Clause 76, wherein the plurality of polymer nanoparticles are amphiphilic.

[0277] Clause 78. The method according to Clause 76 or 77, wherein the plurality of polymer nanoparticles are poly(ethylene glycol)-block-poly(lactic acid) (PEG-PLA) nanoparticles.

[0278] Clause 79. The method according to any one of Clauses 47 to 78, wherein the concentration of the polymer in the dynamic hydrogel is 0.5 wt% to 5 wt%, 0.5 wt% to 4 wt%, 0.5 wt% to 3 wt%, 0.5 wt% to 2 wt%, 0.5 wt% to 1 wt%, 1 wt% to 5 wt%, 1 wt% to 4 wt%, 1 wt% to 3 wt%, 1 wt% to 2 wt%, 2 wt% to 5 wt%, 2 wt% to 4 wt%, 2 wt% to 3 wt%, 3 wt% to 5 wt%, 3 wt% to 4 wt%, or 4 wt% to 5 wt%.

[0279] Clause 80. The method according to any one of Clauses 47 to 79, wherein the concentration of nanoparticles in the dynamic hydrogel is 1 wt% to 12 wt%, 1 wt% to 10 wt%, 1 wt% to 8 wt%, 1 wt% to 5 wt%, 1 wt% to 3 wt%, 3 wt% to 12 wt%, 3 wt% to 10 wt%, 3 wt% to 8 wt%, 3 wt% to 5 wt%, 5 wt% to 12 wt%, 5 wt% to 10 wt%, 5 wt% to 8 wt%, 8 wt% to 12 wt%, 8 wt% to 10 wt%, or 10 wt% to 12 wt%.

[0280] Clause 81. The method according to any one of Clauses 47 to 80, wherein when measured at 25°C at an angular frequency of 0.1 rad / s to 100 rad / s in the linear viscoelastic region of the dynamic hydrogel, the storage modulus of the dynamic hydrogel encapsulating the acylated peptide is 10 Pa to 1000 Pa or 50 Pa to 500 Pa.

[0281] Clause 82. The method according to any one of Clauses 47 to 81, wherein the yield stress of the dynamic hydrogel encapsulating the acylated peptide is from 1 Pa to 500 Pa or from 20 Pa to 200 Pa when measured at 25°C.

[0282] Clause 83. The method according to any one of Clauses 47 to 82, wherein when at 25°C for 1000 s -1 When measured at a shear rate of 100 mPa-s to 1000 mPa-s, the viscosity of the dynamic hydrogel encapsulating the acylated peptide was 100 mPa-s to 1000 mPa-s.

[0283] Clause 84. The method according to any one of Clauses 47 to 83, wherein, after administration to a subject, the composition delivers the acylated peptide to the subject for a treatment period of at least 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 28 days, 35 days, 40 days, 50 days, 60 days, 70 days, 80 days, 90 days, 100 days, 110 days, 120 days, 150 days, 180 days, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or 1 year.

[0284] Clause 85. The method according to any one of Clauses 47 to 84, wherein, after administration to a subject, the composition delivers the acylated peptide to the subject at a rate of 0.25 mg / week to 25 mg / week, 0.5 mg / week to 20 mg / week, 0.5 mg / week to 2 mg / week, 0.5 mg / week to 1.5 mg / week, 1 mg / week to 2 mg / week, 2 mg / week to 5 mg / week, 5 mg / week to 15 mg / week, 5 mg / week to 10 mg / week, 10 mg / week to 20 mg / week, 10 mg / week to 15 mg / week, 12 mg / week to 25 mg / week, 12 mg / week to 15 mg / week, 15 mg / week to 25 mg / week, 15 mg / week to 20 mg / week, or 20 mg / week to 25 mg / week.

[0285] Clause 86. The method according to any one of Clauses 47 to 85, wherein, after administration to a subject, the composition produces in the serum of the subject an average concentration of the acylated peptide of 50 ng / mL to 500 ng / mL, 100 ng / mL to 300 ng / mL, or 150 ng / mL to 250 ng / mL.

[0286] Clause 87. The method according to any one of Clauses 47 to 86, wherein, after administration to a subject, the composition produces less than or equal to 2000 ng / mL of C in the serum of the subject. max The acylated peptide.

[0287] Clause 88. The method according to any one of Clauses 47 to 87, wherein the composition is administered via subcutaneous injection.

[0288] Clause 89. The method according to any one of Clauses 47 to 88, wherein the disease or condition includes diabetes or diabetes-related conditions.

[0289] Clause 90. The method described in Clause 89, wherein the diabetes or diabetes-related condition includes prediabetes, type 1 diabetes, type 2 diabetes, hyperglycemia, or impaired glucose tolerance.

[0290] Clause 91. The method according to any one of Clauses 47 to 90, wherein the disease or condition includes one or more of obesity, overweight, eating disorders, or obstructive sleep apnea.

[0291] Clause 92. The method according to any one of Clauses 47 to 91, wherein the disease or condition includes one or more of the following: cardiovascular disease, liver disease, nervous system disease or neurodegenerative disease, inflammatory disease, kidney disease, bone disease, hormonal disease or gastrointestinal disease.

[0292] Clause 93. A composition for treating a disease or condition, said composition comprising: a dynamic hydrogel comprising a polymer and a plurality of nanoparticles, wherein the polymer is non-covalently crosslinked with the plurality of nanoparticles; and an incretin mimicry encapsulated by said dynamic hydrogel.

[0293] Clause 94. The composition according to Clause 93, wherein the incretin mimic comprises a glucagon-like peptide-1 receptor agonist (GLP-1 RA).

[0294] Clause 95. The composition according to Clause 94, wherein the GLP-1 RA comprises exenatide, exenatide-LAR, lixinatide, liraglutide, smegglutide, abiglutide, duraglutide, ipegglutide, enoglutide, ifenopeptide, cotoditide, mastodide, BI 45690, telpogide, LY3493269, VK2735, CT-868, AMG133, or retaglutide.

[0295] Clause 96. The composition according to Clause 94, wherein the GLP-1 RA comprises liraglutide, smegglutide, enoglutide, cotodextrin, mastodextrin, telpogextrin, or retaglutide.

[0296] Clause 97. The composition according to any one of Clauses 94 to 96, wherein the GLP-1 RA comprises a peptide having a sequence having at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, or 99% sequence identity with any one of SEQ ID NO: 1 or SEQ ID NO: 2.

[0297] Clause 98. The composition according to any one of Clauses 93 to 97, wherein the incretin mimic binds only to the GLP-1 receptor.

[0298] Clause 99. The composition according to any one of Clauses 93 to 97, wherein the incretin mimic binds to a GLP-1 receptor and at least one additional receptor.

[0299] Clause 100. The composition according to Clause 99, wherein the at least one additional receptor comprises one or more of glucagon receptor or gastric inhibitory peptide (GIP) receptor.

[0300] Clause 101. The composition according to any one of Clauses 93 to 100, wherein the incretin mimic comprises an acylated peptide.

[0301] Clause 102. The composition according to Clause 101, wherein the acylated peptide comprises a lipophilic substituent.

[0302] Clause 103. The composition according to Clause 102, wherein the acylated peptide is encapsulated in the dynamic hydrogel via the interaction between the lipophilic substituent and the dynamic hydrogel.

[0303] Clause 104. The composition according to Clause 103, wherein the plurality of nanoparticles have a hydrophobic surface that interacts with the lipophilic substituents of the dynamic hydrogel.

[0304] Clause 105. The composition according to any one of Clauses 102 to 104, wherein the lipophilic substituent comprises an acyl group of a fatty acid.

[0305] Clause 106. The composition according to Clause 105, wherein the fatty acid is a C4 fatty acid, C6 fatty acid, C8 fatty acid, C10 fatty acid, C12 fatty acid, C14 fatty acid, C15 fatty acid, C16 fatty acid, C17 fatty acid, C18 fatty acid, C20 fatty acid, C22 fatty acid, C24 fatty acid, C26 fatty acid, C28 fatty acid, C30 fatty acid, C32 fatty acid, C34 fatty acid, C36 fatty acid, or C38 fatty acid.

[0306] Clause 107. The composition according to any one of Clauses 102 to 106 further comprises a binding agent that interacts with the lipophilic substituent to physically encapsulate the acylated peptide in the dynamic hydrogel.

[0307] Clause 108. The composition according to Clause 107, wherein the binder comprises albumin.

[0308] Clause 109. The composition according to Clause 107 or 108, wherein the binder is larger than the mesh size of the dynamic hydrogel.

[0309] Clause 110. The composition according to any one of Clauses 101 to 109, wherein the acylated peptide is smaller than the pore size of the dynamic hydrogel.

[0310] Clause 111. The composition according to any one of Clauses 101 to 110 further comprises a dispersant that inhibits the aggregation of the acylated peptide.

[0311] Clause 112. The composition according to Clause 111, wherein the dispersant comprises one or more of a surfactant or an osmotic pressure regulator.

[0312] Clause 113. The composition according to Clause 112, wherein the dispersant comprises the surfactant, and the surfactant comprises one or more of polysorbate, sorbitol fatty acid ester, poloxamer, polyoxyethylene alkyl ether, alkyl sulfate or a salt thereof, fatty acid, fatty acid alcohol or phospholipid.

[0313] Clause 114. The composition according to Clause 112 or 113, wherein the dispersant comprises the osmotic pressure regulator, and the osmotic pressure regulator comprises one or more of propylene glycol, glycerol, or mannitol.

[0314] Clause 115. The composition according to any one of Clauses 112 to 114, wherein the concentration of the dispersant in the composition is 0.1 mg / mL to 5 mg / mL, 0.5 mg / mL to 1.5 mg / mL, 1 mg / mL to 2 mg / mL, 1 mg / mL to 50 mg / mL, 5 mg / mL to 40 mg / mL, 10 mg / mL to 30 mg / mL, or 16 mg / mL to 26 mg / mL.

[0315] Clause 116. The composition according to any one of Clauses 93 to 100, wherein the incretin mimic comprises a peptide conjugated to a protein or protein fragment.

[0316] Clause 117. The composition according to any one of Clauses 93 to 100, wherein the incretin mimic comprises a peptide encapsulated in microparticles.

[0317] Clause 118. The composition according to any one of Clauses 93 to 117, wherein the composition comprises 1 mg to 250 mg of the incretin mimic.

[0318] Clause 119. The composition according to any one of Clauses 93 to 118, wherein the concentration of the incretin mimic in the composition is from 1 mg / mL to 200 mg / mL.

[0319] Clause 120. The composition according to any one of Clauses 93 to 119, wherein said polymer comprises a hydrophobically modified polysaccharide.

[0320] Clause 121. The composition according to Clause 120, wherein the hydrophobically modified polysaccharide comprises a hydrophobically modified cellulose derivative.

[0321] Clause 122. The composition according to Clause 121, wherein the hydrophobically modified cellulose derivative is dodecyl modified hydroxypropyl methylcellulose (HPMC-C12).

[0322] Clause 123. The composition according to any one of Clauses 93 to 122, wherein the plurality of nanoparticles comprises a plurality of polymer nanoparticles.

[0323] Clause 124. The composition according to Clause 123, wherein the plurality of polymer nanoparticles are amphiphilic.

[0324] Clause 125. The composition according to Clause 123 or 124, wherein the plurality of polymer nanoparticles are poly(ethylene glycol)-block-poly(lactic acid) (PEG-PLA) nanoparticles.

[0325] Clause 126. The composition according to any one of Clauses 93 to 125, wherein the concentration of the polymer in the dynamic hydrogel is 0.5 wt% to 5 wt%, 0.5 wt% to 4 wt%, 0.5 wt% to 3 wt%, 0.5 wt% to 2 wt%, 0.5 wt% to 1 wt%, 1 wt% to 5 wt%, 1 wt% to 4 wt%, 1 wt% to 3 wt%, 1 wt% to 2 wt%, 2 wt% to 5 wt%, 2 wt% to 4 wt%, 2 wt% to 3 wt%, 3 wt% to 5 wt%, 3 wt% to 4 wt%, or 4 wt% to 5 wt%.

[0326] Clause 127. The composition according to any one of Clauses 93 to 126, wherein the concentration of nanoparticles in the dynamic hydrogel is 1 wt% to 12 wt%, 1 wt% to 10 wt%, 1 wt% to 8 wt%, 1 wt% to 5 wt%, 1 wt% to 3 wt%, 3 wt% to 12 wt%, 3 wt% to 10 wt%, 3 wt% to 8 wt%, 3 wt% to 5 wt%, 5 wt% to 12 wt%, 5 wt% to 10 wt%, 5 wt% to 8 wt%, 8 wt% to 12 wt%, 8 wt% to 10 wt%, or 10 wt% to 12 wt%.

[0327] Clause 128. The composition according to any one of Clauses 93 to 127, wherein the storage modulus of the dynamic hydrogel encapsulating the incretin mimic is 10 Pa to 1000 Pa or 50 Pa to 500 Pa when measured at 25°C at an angular frequency of 0.1 rad / s to 100 rad / s in the linear viscoelastic region of the dynamic hydrogel.

[0328] Clause 129. The composition according to any one of Clauses 93 to 128, wherein the yield stress of the dynamic hydrogel encapsulating the incretin mimic is from 1 Pa to 500 Pa or from 20 Pa to 200 Pa when measured at 25°C.

[0329] Clause 130. The composition according to any one of Clauses 93 to 129, wherein when at 25°C for 1000 s -1 When measured at a shear rate of 100 mPa-s to 1000 mPa-s, the viscosity of the dynamic hydrogel encapsulating the incretin mimic was 100 mPa-s to 1000 mPa-s.

[0330] Clause 131. The composition according to any one of Clauses 93 to 130, wherein, after administration to a subject, the composition delivers the incretin mimic to the subject for a treatment period of at least 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 28 days, 35 days, 40 days, 50 days, 60 days, 70 days, 80 days, 90 days, 100 days, 110 days, 120 days, 150 days, 180 days, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or 1 year.

[0331] Clause 132. The composition according to any one of Clauses 93 to 131, wherein, after administration to the subject, the composition delivers the incretin mimic to the subject at a rate of 0.25 mg / week to 25 mg / week, 0.5 mg / week to 20 mg / week, 0.5 mg / week to 2 mg / week, 0.5 mg / week to 1.5 mg / week, 1 mg / week to 2 mg / week, 2 mg / week to 5 mg / week, 5 mg / week to 15 mg / week, 5 mg / week to 10 mg / week, 10 mg / week to 20 mg / week, 10 mg / week to 15 mg / week, 12 mg / week to 25 mg / week, 12 mg / week to 15 mg / week, 15 mg / week to 25 mg / week, 15 mg / week to 20 mg / week, or 20 mg / week to 25 mg / week.

[0332] Clause 133. The composition according to any one of Clauses 93 to 132, wherein, after administration to a subject, the composition produces in the serum of the subject an average concentration of the incretin mimic at a rate of 50 ng / mL to 500 ng / mL, 100 ng / mL to 300 ng / mL, or 150 ng / mL to 250 ng / mL.

[0333] Clause 134. The composition according to any one of Clauses 93 to 133, wherein, after administration to a subject, the composition produces less than or equal to 2000 ng / mL of C in the serum of the subject. max The aforementioned incretin mimicry.

[0334] Clause 135. The composition according to any one of Clauses 93 to 134, wherein the composition is configured for administration via subcutaneous injection.

[0335] Clause 136. The composition according to any one of Clauses 93 to 135, wherein the disease or condition includes diabetes or diabetes-related conditions.

[0336] Clause 137. The composition according to Clause 136, wherein the diabetes or diabetes-related condition includes prediabetes, type 1 diabetes, type 2 diabetes, hyperglycemia, or impaired glucose tolerance.

[0337] Clause 138. The composition according to any one of Clauses 93 to 137, wherein the disease or condition includes one or more of obesity, overweight, eating disorders, or obstructive sleep apnea.

[0338] Clause 139. The composition according to any one of Clauses 93 to 138, wherein the disease or condition includes one or more of the following: cardiovascular disease, liver disease, nervous system disease or neurodegenerative disease, inflammatory disease, kidney disease, bone disease, hormonal disease or gastrointestinal disease.

[0339] Clause 140. A method of treating a disease or condition, the method comprising: applying a composition to a subject, wherein the composition comprises: a dynamic hydrogel comprising a polymer and a plurality of nanoparticles, wherein the polymer is non-covalently crosslinked with the plurality of nanoparticles; and an incretin mimic, the incretin mimic being encapsulated by the dynamic hydrogel.

[0340] Clause 141. The method according to Clause 140, wherein the incretin mimic comprises a glucagon-like peptide-1 receptor agonist (GLP-1 RA).

[0341] Clause 142. The method according to Clause 141, wherein the GLP-1 RA comprises exenatide, exenatide-LAR, lixinatide, liraglutide, smegglutide, abiglutide, duraglutide, ipegglutide, enoglutide, ifenopeptide, cotoditide, mastodide, BI 45690, telpogide, LY3493269, VK2735, CT-868, AMG133, or retaglutide.

[0342] Clause 143. The method according to Clause 141, wherein the GLP-1 RA comprises liraglutide, smegglutide, enoglutide, cotodextrin, mastodextrin, telpogextrin, or retaglutide.

[0343] Clause 144. The method according to any one of Clauses 140 to 143, wherein the GLP-1 RA comprises a peptide having a sequence having at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, or 99% sequence identity with either SEQ ID NO: 1 or SEQ ID NO: 2.

[0344] Clause 145. The method according to any one of Clauses 140 to 144, wherein the incretin mimic binds only to the GLP-1 receptor.

[0345] Clause 146. The method according to any one of Clauses 140 to 144, wherein the incretin mimic binds to the GLP-1 receptor and at least one additional receptor.

[0346] Clause 147. The method according to Clause 146, wherein the at least one additional receptor comprises one or more of glucagon receptor or gastric inhibitory peptide (GIP) receptor.

[0347] Clause 148. The method according to any one of Clauses 140 to 147, wherein the incretin mimic comprises an acylated peptide.

[0348] Clause 149. The method according to Clause 148, wherein the acylated peptide comprises a lipophilic substituent.

[0349] Clause 150. The method according to Clause 149, wherein the acylated peptide is encapsulated in the dynamic hydrogel via an interaction between the lipophilic substituent and the dynamic hydrogel.

[0350] Clause 151. The method according to Clause 150, wherein the plurality of nanoparticles have a hydrophobic surface that interacts with the lipophilic substituents of the dynamic hydrogel.

[0351] Clause 152. The method according to any one of Clauses 149 to 151, wherein the lipophilic substituent comprises an acyl group of a fatty acid.

[0352] Clause 153. The method according to Clause 152, wherein the fatty acid is a C4 fatty acid, C6 fatty acid, C8 fatty acid, C10 fatty acid, C12 fatty acid, C14 fatty acid, C15 fatty acid, C16 fatty acid, C17 fatty acid, C18 fatty acid, C20 fatty acid, C22 fatty acid, C24 fatty acid, C26 fatty acid, C28 fatty acid, C30 fatty acid, C32 fatty acid, C34 fatty acid, C36 fatty acid, or C38 fatty acid.

[0353] Clause 154. The method according to any one of Clauses 149 to 153, wherein the composition further comprises a binding agent that interacts with the lipophilic substituent to physically encapsulate the acylated peptide in the dynamic hydrogel.

[0354] Clause 155. The method according to Clause 154, wherein the binding agent comprises albumin.

[0355] Clause 156. The method according to Clause 154 or 155, wherein the binder is larger than the mesh size of the dynamic hydrogel.

[0356] Clause 157. The method according to any one of Clauses 148 to 156, wherein the acylated peptide is smaller than the pore size of the dynamic hydrogel.

[0357] Clause 158. The composition further comprises a dispersant that inhibits the aggregation of the acylated peptide according to any one of Clauses 148 to 157.

[0358] Clause 159. The method according to Clause 158, wherein the dispersant comprises one or more of a surfactant or an osmotic pressure regulator.

[0359] Clause 160. The method according to Clause 159, wherein the dispersant comprises the surfactant, and the surfactant comprises one or more of polysorbate, sorbitol fatty acid ester, poloxamer, polyoxyethylene alkyl ether, alkyl sulfate or its salt, fatty acid, fatty acid alcohol or phospholipid.

[0360] Clause 161. The method according to Clause 159 or 160, wherein the dispersant comprises the osmotic pressure regulator, and the osmotic pressure regulator comprises one or more of propylene glycol, glycerol, or mannitol.

[0361] Clause 162. The method according to any one of Clauses 159 to 161, wherein the concentration of the dispersant in the composition is 0.1 mg / mL to 5 mg / mL, 0.5 mg / mL to 1.5 mg / mL, 1 mg / mL to 2 mg / mL, 1 mg / mL to 50 mg / mL, 5 mg / mL to 40 mg / mL, 10 mg / mL to 30 mg / mL, or 16 mg / mL to 26 mg / mL.

[0362] Clause 163. The method according to any one of Clauses 140 to 147, wherein the incretin mimic comprises a peptide conjugated to a protein or protein fragment.

[0363] Clause 164. The method according to any one of Clauses 140 to 147, wherein the incretin mimic comprises a peptide encapsulated in microparticles.

[0364] Clause 165. The method according to any one of Clauses 140 to 164, wherein the composition comprises 1 mg to 250 mg of the incretin mimic.

[0365] Clause 166. The method according to any one of Clauses 140 to 165, wherein the concentration of the incretin mimic in the composition is from 1 mg / mL to 200 mg / mL.

[0366] Clause 167. The method according to any one of Clauses 140 to 166, wherein said polymer comprises a hydrophobically modified polysaccharide.

[0367] Clause 168. The method according to Clause 167, wherein the hydrophobically modified polysaccharide comprises a hydrophobically modified cellulose derivative.

[0368] Clause 169. The method according to Clause 168, wherein the hydrophobically modified cellulose derivative is dodecyl modified hydroxypropyl methylcellulose (HPMC-C12).

[0369] Clause 170. The method according to any one of Clauses 140 to 169, wherein the plurality of nanoparticles comprises a plurality of polymer nanoparticles.

[0370] Clause 171. The method according to Clause 170, wherein the plurality of polymer nanoparticles are amphiphilic.

[0371] Clause 172. The method according to Clause 170 or 171, wherein the plurality of polymer nanoparticles are poly(ethylene glycol)-block-poly(lactic acid) (PEG-PLA) nanoparticles.

[0372] Clause 173. The method according to any one of Clauses 140 to 172, wherein the concentration of the polymer in the dynamic hydrogel is 0.5 wt% to 5 wt%, 0.5 wt% to 4 wt%, 0.5 wt% to 3 wt%, 0.5 wt% to 2 wt%, 0.5 wt% to 1 wt%, 1 wt% to 5 wt%, 1 wt% to 4 wt%, 1 wt% to 3 wt%, 1 wt% to 2 wt%, 2 wt% to 5 wt%, 2 wt% to 4 wt%, 2 wt% to 3 wt%, 3 wt% to 5 wt%, 3 wt% to 4 wt%, or 4 wt% to 5 wt%.

[0373] Clause 174. The method according to any one of Clauses 140 to 173, wherein the concentration of nanoparticles in the dynamic hydrogel is 1 wt% to 12 wt%, 1 wt% to 10 wt%, 1 wt% to 8 wt%, 1 wt% to 5 wt%, 1 wt% to 3 wt%, 3 wt% to 12 wt%, 3 wt% to 10 wt%, 3 wt% to 8 wt%, 3 wt% to 5 wt%, 5 wt% to 12 wt%, 5 wt% to 10 wt%, 5 wt% to 8 wt%, 8 wt% to 12 wt%, 8 wt% to 10 wt%, or 10 wt% to 12 wt%.

[0374] Clause 175. The method according to any one of Clauses 140 to 174, wherein the storage modulus of the dynamic hydrogel encapsulating the incretin mimic is 10 Pa to 1000 Pa or 50 Pa to 500 Pa when measured at 25°C at an angular frequency of 0.1 rad / s to 100 rad / s in the linear viscoelastic region of the dynamic hydrogel.

[0375] Clause 176. The method according to any one of Clauses 140 to 175, wherein the yield stress of the dynamic hydrogel encapsulating the incretin mimic is 1 Pa to 500 Pa or 20 Pa to 200 Pa when measured at 25°C.

[0376] Clause 177. The method according to any one of Clauses 140 to 176, wherein when at 25°C for 1000 s -1 When measured at a shear rate of 100 mPa-s to 1000 mPa-s, the viscosity of the dynamic hydrogel encapsulating the incretin mimic was 100 mPa-s to 1000 mPa-s.

[0377] Clause 178. The method according to any one of Clauses 140 to 177, wherein, after administration to a subject, the composition is delivered to the subject for a treatment period of at least 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 28 days, 35 days, 40 days, 50 days, 60 days, 70 days, 80 days, 90 days, 100 days, 110 days, 120 days, 150 days, 180 days, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or 1 year.

[0378] Clause 179. The method according to any one of Clauses 140 to 178, wherein, after administration to a subject, the composition delivers the incretin mimic to the subject at a rate of 0.25 mg / week to 25 mg / week, 0.5 mg / week to 20 mg / week, 0.5 mg / week to 2 mg / week, 0.5 mg / week to 1.5 mg / week, 1 mg / week to 2 mg / week, 2 mg / week to 5 mg / week, 5 mg / week to 15 mg / week, 5 mg / week to 10 mg / week, 10 mg / week to 20 mg / week, 10 mg / week to 15 mg / week, 12 mg / week to 25 mg / week, 12 mg / week to 15 mg / week, 15 mg / week to 25 mg / week, 15 mg / week to 20 mg / week, or 20 mg / week to 25 mg / week.

[0379] Clause 180. The method according to any one of Clauses 140 to 179, wherein, after administration to a subject, the composition produces in the serum of the subject an average concentration of the incretin mimic at a rate of 50 ng / mL to 500 ng / mL, 100 ng / mL to 300 ng / mL, or 150 ng / mL to 250 ng / mL.

[0380] Clause 181. The method according to any one of Clauses 140 to 180, wherein, after administration to a subject, the composition produces less than or equal to 2000 ng / mL of C in the serum of the subject. max The aforementioned incretin mimicry.

[0381] Clause 182. The method according to any one of Clauses 140 to 181, wherein the composition is administered via subcutaneous injection.

[0382] Clause 183. The method according to any one of Clauses 140 to 182, wherein the disease or condition includes diabetes or diabetes-related conditions.

[0383] Clause 184. The method described in Clause 183, wherein the diabetes or diabetes-related condition includes prediabetes, type 1 diabetes, type 2 diabetes, hyperglycemia, or impaired glucose tolerance.

[0384] Clause 185. The method according to any one of Clauses 140 to 184, wherein the disease or condition includes one or more of obesity, overweight, eating disorders or obstructive sleep apnea.

[0385] Clause 186. The method according to any one of Clauses 140 to 185, wherein the disease or condition includes one or more of the following: cardiovascular disease, liver disease, nervous system disease or neurodegenerative disease, inflammatory disease, kidney disease, bone disease, hormonal disease or gastrointestinal disease.

[0386] in conclusion

[0387] While numerous embodiments have been described above regarding compositions and methods for delivering incretin mimics, this technology can be applied to other applications and / or other methods, such as delivering other types of therapeutic peptides. Furthermore, embodiments other than those described herein are also within the scope of this technology. Additionally, several other embodiments of this technology may have configurations, components, or processes different from those described herein. Therefore, those skilled in the art will accordingly understand that the technology can have other embodiments with additional elements, or the technology may have embodiments without the above references. Figures 1A-17Other implementations of several features shown and described.

[0388] The description of embodiments of this technology is not intended to be exhaustive or to limit the technology to the precise forms disclosed above. Where context permits, singular or plural terms may also include plural or singular terms, respectively. Although specific embodiments and examples of the technology have been described above for illustrative purposes, as those skilled in the art will recognize, various equivalent modifications are possible within the scope of the technology. For example, although the steps are presented in a given order, alternative embodiments may perform the steps in a different order. The various embodiments described herein can also be combined to provide other embodiments.

[0389] As used herein, the terms “usually,” “substantially,” “about,” and similar terms are used as approximations rather than as terms of degree and are intended to account for inherent variations in measured or calculated values ​​that would be recognized by one of ordinary skill in the art. As used herein, the terms “about” and “approximately” with respect to numbers can include numbers falling within the range of 10%, 5%, or 1% in either direction (greater than or less than) of the number, unless otherwise stated or apparent from the context (unless such a number would exceed 100% of the possible value).

[0390] Furthermore, when referring to a list of two or more items, unless the word "or" is explicitly limited to meaning only a single item excluding other items, its use in such a list should be interpreted as including: (a) any single item in the list, (b) all items in the list, or (c) any combination of items in the list. As used herein, the phrase "and / or" in "A and / or B" means only A, only B, and A and B. Furthermore, the term "including / contains" is used throughout to mean at least the stated feature, such that no further number of the same feature and / or additional features of an additional type are excluded.

[0391] As used herein, the term “object” can refer to any animal, including but not limited to humans and non-human animals (e.g., dogs, cats, cows, horses, sheep, pigs, poultry, fish, crustaceans, etc.).

[0392] As used herein, in the context of two or more nucleic acid or polypeptide sequences, the term "identity" percentage can refer to two or more sequences or subsequences that, when compared and aligned for maximum correspondence, have a specified percentage of identical nucleotide or amino acid residues, as measured by visual inspection, such as using one of the sequence comparison algorithms described below (e.g., BLASTP and BLASTN or other algorithms available to a technician). Depending on the application, the "identity" percentage may be present in a region of the sequences being compared, such as in a functional domain, or alternatively, across the full length of the two sequences to be compared. For sequence comparisons, typically one sequence is used as a reference sequence to be compared with the test sequence. When using a sequence comparison algorithm, the test sequence and reference sequence are input into a computer, and if necessary, the coordinates of the subsequences are specified, along with the sequence algorithm program parameters. The sequence comparison algorithm then calculates the sequence identity percentage of the test sequence relative to the reference sequence based on the specified program parameters. For the purposes of this paper, the BLAST algorithm described in Altschul et al. (J. Mol. Biol. 215:403-410 (1990)) can be used for identity percentage and sequence similarity analysis. The software used for BLAST analysis is publicly available from the National Center for Biotechnology Information.

[0393] In the event of any conflict between this disclosure and any material incorporated herein by reference, this disclosure shall prevail.

[0394] It should also be understood that, for illustrative purposes, specific embodiments have been described herein, but various modifications may be made without departing from the art. Furthermore, while advantages associated with certain embodiments of the art have been described in the context of these embodiments, other embodiments may also exhibit such advantages, and not all embodiments must exhibit such advantages that fall within the scope of the art. Therefore, this disclosure and related art may cover other embodiments not expressly shown or described herein.

Claims

1. A composition for treating a disease or condition, said composition comprising: Dynamic hydrogel comprising a polymer and a plurality of nanoparticles, wherein the polymer is non-covalently crosslinked with the plurality of nanoparticles; and Acylated peptides, which are encapsulated by the dynamic hydrogel.

2. The composition according to claim 1, wherein the acylated peptide comprises a lipophilic substituent.

3. The composition of claim 2, wherein the acylated peptide is encapsulated in the dynamic hydrogel via the interaction between the lipophilic substituent and the dynamic hydrogel.

4. The composition of claim 3, wherein the plurality of nanoparticles have a hydrophobic surface that interacts with the lipophilic substituents of the dynamic hydrogel.

5. The composition according to any one of claims 2 to 4, wherein the lipophilic substituent comprises an acyl group of a fatty acid.

6. The composition according to claim 5, wherein the fatty acid is a C4 fatty acid, C6 fatty acid, C8 fatty acid, C10 fatty acid, C12 fatty acid, C14 fatty acid, C15 fatty acid, C16 fatty acid, C17 fatty acid, C18 fatty acid, C20 fatty acid, C22 fatty acid, C24 fatty acid, C26 fatty acid, C28 fatty acid, C30 fatty acid, C32 fatty acid, C34 fatty acid, C36 fatty acid, or C38 fatty acid.

7. The composition according to any one of claims 2 to 6, further comprising a binding agent that interacts with the lipophilic substituent to physically encapsulate the acylated peptide in the dynamic hydrogel.

8. The composition of claim 7, wherein the binder comprises albumin.

9. The composition according to claim 7 or 8, wherein the binder is larger than the mesh size of the dynamic hydrogel.

10. The composition according to any one of claims 1 to 9, wherein the acylated peptide is smaller than the pore size of the dynamic hydrogel.

11. The composition according to any one of claims 1 to 10, further comprising a dispersant that inhibits the aggregation of the acylated peptide.

12. The composition of claim 11, wherein the dispersant comprises one or more of a surfactant or an osmotic pressure regulator.

13. The composition of claim 12, wherein the dispersant comprises the surfactant, and the surfactant comprises one or more of polysorbate, sorbitol fatty acid ester, poloxamer, polyoxyethylene alkyl ether, alkyl sulfate, fatty acid, fatty acid alcohol or phospholipid.

14. The composition according to claim 12 or 13, wherein the dispersant comprises the osmotic pressure regulator, and the osmotic pressure regulator comprises one or more of propylene glycol, glycerol, or mannitol.

15. The composition according to any one of claims 12 to 14, wherein the concentration of the dispersant in the composition is 0.1 mg / mL to 5 mg / mL, 0.5 mg / mL to 1.5 mg / mL, 1 mg / mL to 2 mg / mL, 1 mg / mL to 50 mg / mL, 5 mg / mL to 40 mg / mL, 10 mg / mL to 30 mg / mL, or 16 mg / mL to 26 mg / mL.

16. The composition according to any one of claims 1 to 15, wherein the acylated peptide comprises an incretin mimic.

17. The composition of claim 16, wherein the incretin mimic comprises a glucagon-like peptide-1 receptor agonist (GLP-1 RA).

18. The composition of claim 17, wherein the GLP-1 RA comprises one or more of liraglutide, smegglutide, enoglutide, cotodextrin, mastodextrin, telpogextrin, or retaglutide.

19. The composition according to any one of claims 16 to 18, wherein the incretin mimic binds only to the GLP-1 receptor.

20. The composition according to any one of claims 16 to 18, wherein the incretin mimic binds to a GLP-1 receptor and at least one additional receptor.

21. The composition of claim 20, wherein the at least one additional receptor comprises one or more of glucagon receptor or gastric inhibitory peptide (GIP) receptor.

22. The composition according to any one of claims 1 to 15, wherein the acylated peptide comprises an analog of a proglucagon-derived peptide.

23. The composition according to any one of claims 1 to 15, wherein the acylated peptide comprises an amylin analogue.

24. The composition according to any one of claims 1 to 15, wherein the acylated peptide comprises an insulin analog.

25. The composition according to any one of claims 1 to 24, wherein the composition comprises 1 mg to 250 mg of the acylated peptide.

26. The composition according to any one of claims 1 to 25, wherein the concentration of the acylated peptide in the composition is from 1 mg / mL to 200 mg / mL.

27. The composition according to any one of claims 1 to 26, wherein the polymer comprises a hydrophobically modified polysaccharide.

28. The composition of claim 27, wherein the hydrophobically modified polysaccharide comprises a hydrophobically modified cellulose derivative.

29. The composition according to claim 28, wherein the hydrophobically modified cellulose derivative is dodecyl modified hydroxypropyl methylcellulose (HPMC-C12).

30. The composition according to any one of claims 1 to 29, wherein the plurality of nanoparticles comprises a plurality of polymer nanoparticles.

31. The composition of claim 30, wherein the plurality of polymer nanoparticles are amphiphilic.

32. The composition according to claim 30 or 31, wherein the plurality of polymer nanoparticles are poly(ethylene glycol)-block-poly(lactic acid) (PEG-PLA) nanoparticles.

33. The composition according to any one of claims 1 to 32, wherein the concentration of the polymer in the dynamic hydrogel is 0.5 wt% to 5 wt%, 0.5 wt% to 4 wt%, 0.5 wt% to 3 wt%, 0.5 wt% to 2 wt%, 0.5 wt% to 1 wt%, 1 wt% to 5 wt%, 1 wt% to 4 wt%, 1 wt% to 3 wt%, 1 wt% to 2 wt%, 2 wt% to 5 wt%, 2 wt% to 4 wt%, 2 wt% to 3 wt%, 3 wt% to 5 wt%, 3 wt% to 4 wt%, or 4 wt% to 5 wt%.

34. The composition according to any one of claims 1 to 33, wherein the concentration of the nanoparticles in the dynamic hydrogel is 1 wt% to 12 wt%, 1 wt% to 10 wt%, 1 wt% to 8 wt%, 1 wt% to 5 wt%, 1 wt% to 3 wt%, 3 wt% to 12 wt%, 3 wt% to 10 wt%, 3 wt% to 8 wt%, 3 wt% to 5 wt%, 5 wt% to 12 wt%, 5 wt% to 10 wt%, 5 wt% to 8 wt%, 8 wt% to 12 wt%, 8 wt% to 10 wt%, or 10 wt% to 12 wt%.

35. The composition according to any one of claims 1 to 34, wherein the storage modulus of the dynamic hydrogel encapsulating the acylated peptide is 10 Pa to 1000 Pa or 50 Pa to 500 Pa when measured at 25°C at an angular frequency of 0.1 rad / s to 100 rad / s in the linear viscoelastic region of the dynamic hydrogel.

36. The composition according to any one of claims 1 to 35, wherein the yield stress of the dynamic hydrogel encapsulating the acylated peptide is 1 Pa to 500 Pa or 20 Pa to 200 Pa when measured at 25°C.

37. The composition according to any one of claims 1 to 36, wherein when at 25°C for 1000 s -1 When measured at a shear rate of 100 mPa-s to 1000 mPa-s, the viscosity of the dynamic hydrogel encapsulating the acylated peptide was 100 mPa-s to 1000 mPa-s.

38. The composition according to any one of claims 1 to 37, wherein, After administration to a subject, the composition delivers the acylated peptide to the subject for a treatment period of at least 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 28 days, 35 days, 40 days, 50 days, 60 days, 70 days, 80 days, 90 days, 100 days, 110 days, 120 days, 150 days, 180 days, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or 1 year.

39. The composition according to any one of claims 1 to 38, wherein, After administration to a subject, the composition delivers the acylated peptide to the subject at rates of 0.25 mg / week to 25 mg / week, 0.5 mg / week to 20 mg / week, 0.5 mg / week to 2 mg / week, 0.5 mg / week to 1.5 mg / week, 1 mg / week to 2 mg / week, 2 mg / week to 5 mg / week, 5 mg / week to 15 mg / week, 5 mg / week to 10 mg / week, 10 mg / week to 20 mg / week, 10 mg / week to 15 mg / week, 12 mg / week to 25 mg / week, 12 mg / week to 15 mg / week, 15 mg / week to 25 mg / week, 15 mg / week to 20 mg / week, or 20 mg / week to 25 mg / week.

40. The composition according to any one of claims 1 to 39, wherein, Upon administration to a subject, the composition produces an average concentration of the acylated peptide in the subject's serum of 50 ng / mL to 500 ng / mL, 100 ng / mL to 300 ng / mL, or 150 ng / mL to 250 ng / mL.

41. The composition according to any one of claims 1 to 40, wherein, After administration to a subject, the composition produces C in the subject's serum at a concentration of less than or equal to 2000 ng / mL. max The acylated peptide.

42. The composition according to any one of claims 1 to 41, wherein the composition is configured for administration via subcutaneous injection.

43. The composition according to any one of claims 1 to 42, wherein the disease or condition includes diabetes or diabetes-related conditions.

44. The composition of claim 43, wherein the diabetes or diabetes-related condition includes prediabetes, type 1 diabetes, type 2 diabetes, hyperglycemia, or impaired glucose tolerance.

45. The composition according to any one of claims 1 to 44, wherein the disease or condition includes one or more of obesity, overweight, eating disorders, or obstructive sleep apnea.

46. ​​The composition according to any one of claims 1 to 45, wherein the disease or condition includes one or more of the following: cardiovascular disease, liver disease, nervous system disease or neurodegenerative disease, inflammatory disease, kidney disease, bone disease, hormonal disease or gastrointestinal disease.

47. A method for treating a disease or condition, said method comprising: The composition is applied to an object, wherein the composition comprises: Dynamic hydrogel comprising a polymer and multiple nanoparticles, wherein the polymer is non-covalently crosslinked with the multiple nanoparticles, and Acylated peptides, which are encapsulated by the dynamic hydrogel.

48. The method of claim 47, wherein the acylated peptide comprises a lipophilic substituent.

49. The method of claim 48, wherein the acylated peptide is encapsulated in the dynamic hydrogel via the interaction between the lipophilic substituent and the dynamic hydrogel.

50. The method of claim 49, wherein the plurality of nanoparticles have a hydrophobic surface that interacts with the lipophilic substituents of the dynamic hydrogel.

51. The method according to any one of claims 48 to 50, wherein the lipophilic substituent comprises an acyl group of a fatty acid.

52. The method according to claim 51, wherein the fatty acid is a C4 fatty acid, C6 fatty acid, C8 fatty acid, C10 fatty acid, C12 fatty acid, C14 fatty acid, C15 fatty acid, C16 fatty acid, C17 fatty acid, C18 fatty acid, C20 fatty acid, C22 fatty acid, C24 fatty acid, C26 fatty acid, C28 fatty acid, C30 fatty acid, C32 fatty acid, C34 fatty acid, C36 fatty acid, or C38 fatty acid.

53. The method according to any one of claims 48 to 52, wherein the composition further comprises a binding agent that interacts with the lipophilic substituent to physically encapsulate the acylated peptide in the dynamic hydrogel.

54. The method of claim 53, wherein the binding agent comprises albumin.

55. The composition according to claim 53 or 54, wherein the binder is larger than the mesh size of the dynamic hydrogel.

56. The method according to any one of claims 47 to 55, wherein the acylated peptide is smaller than the pore size of the dynamic hydrogel.

57. The method according to any one of claims 47 to 56, wherein the composition further comprises a dispersant that inhibits the aggregation of the acylated peptide.

58. The method of claim 57, wherein the dispersant comprises one or more of a surfactant or an osmotic pressure regulator.

59. The method of claim 58, wherein the dispersant comprises the surfactant, and the surfactant comprises one or more of polysorbate, sorbitol fatty acid ester, poloxamer, polyoxyethylene alkyl ether, alkyl sulfate or its salt, fatty acid, and fatty acid alcohol.

60. The method according to claim 58 or 59, wherein the dispersant comprises the osmotic pressure regulator, and the osmotic pressure regulator comprises one or more of propylene glycol, glycerol, or mannitol.

61. The method according to any one of claims 58 to 60, wherein the concentration of the dispersant in the composition is 0.1 mg / mL to 5 mg / mL, 0.5 mg / mL to 1.5 mg / mL, 1 mg / mL to 2 mg / mL, 1 mg / mL to 50 mg / mL, 5 mg / mL to 40 mg / mL, 10 mg / mL to 30 mg / mL, or 16 mg / mL to 26 mg / mL.

62. The method according to any one of claims 47 to 61, wherein the acylated peptide comprises an incretin mimic.

63. The method of claim 62, wherein the incretin mimic comprises a glucagon-like peptide-1 receptor agonist (GLP-1 RA).

64. The method of claim 63, wherein the GLP-1 RA comprises one or more of liraglutide, smegglutide, enoglutide, cotodextrin, mastodextrin, telpogextrin, or retaglutide.

65. The method according to any one of claims 62 to 64, wherein the incretin mimic binds only to the GLP-1 receptor.

66. The method according to any one of claims 62 to 64, wherein the incretin mimic binds to the GLP-1 receptor and at least one additional receptor.

67. The method of claim 66, wherein the at least one additional receptor comprises one or more of glucagon receptor or gastric inhibitory peptide (GIP) receptor.

68. The method according to any one of claims 47 to 67, wherein the acylated peptide comprises an analog of a proglucagon-derived peptide.

69. The method according to any one of claims 47 to 67, wherein the acylated peptide comprises an amylin analogue.

70. The method according to any one of claims 47-67 or 69, wherein the acylated peptide comprises an insulin analog.

71. The method according to any one of claims 47 to 70, wherein the composition comprises 1 mg to 250 mg of the acylated peptide.

72. The method according to any one of claims 47 to 71, wherein the concentration of the acylated peptide in the composition is from 1 mg / mL to 200 mg / mL.

73. The method according to any one of claims 47 to 72, wherein the polymer comprises a hydrophobically modified polysaccharide.

74. The method of claim 73, wherein the hydrophobically modified polysaccharide comprises a hydrophobically modified cellulose derivative.

75. The method according to claim 74, wherein the hydrophobically modified cellulose derivative is dodecyl modified hydroxypropyl methylcellulose (HPMC-C12).

76. The method according to any one of claims 47 to 75, wherein the plurality of nanoparticles comprises a plurality of polymer nanoparticles.

77. The method of claim 76, wherein the plurality of polymer nanoparticles are amphiphilic.

78. The method according to claim 76 or 77, wherein the plurality of polymer nanoparticles are poly(ethylene glycol)-block-poly(lactic acid) (PEG-PLA) nanoparticles.

79. The method according to any one of claims 47 to 78, wherein the concentration of the polymer in the dynamic hydrogel is 0.5 wt% to 5 wt%, 0.5 wt% to 4 wt%, 0.5 wt% to 3 wt%, 0.5 wt% to 2 wt%, 0.5 wt% to 1 wt%, 1 wt% to 5 wt%, 1 wt% to 4 wt%, 1 wt% to 3 wt%, 1 wt% to 2 wt%, 2 wt% to 5 wt%, 2 wt% to 4 wt%, 2 wt% to 3 wt%, 3 wt% to 5 wt%, 3 wt% to 4 wt%, or 4 wt% to 5 wt%.

80. The method according to any one of claims 47 to 79, wherein the concentration of the nanoparticles in the dynamic hydrogel is 1 wt% to 12 wt%, 1 wt% to 10 wt%, 1 wt% to 8 wt%, 1 wt% to 5 wt%, 1 wt% to 3 wt%, 3 wt% to 12 wt%, 3 wt% to 10 wt%, 3 wt% to 8 wt%, 3 wt% to 5 wt%, 5 wt% to 12 wt%, 5 wt% to 10 wt%, 5 wt% to 8 wt%, 8 wt% to 12 wt%, 8 wt% to 10 wt%, or 10 wt% to 12 wt%.

81. The method according to any one of claims 47 to 80, wherein when measured at 25°C at an angular frequency of 0.1 rad / s to 100 rad / s in the linear viscoelastic region of the dynamic hydrogel, the storage modulus of the dynamic hydrogel encapsulating the acylated peptide is 10 Pa to 1000 Pa or 50 Pa to 500 Pa.

82. The method according to any one of claims 47 to 81, wherein the yield stress of the dynamic hydrogel encapsulating the acylated peptide is 1 Pa to 500 Pa or 20 Pa to 200 Pa when measured at 25°C.

83. The method according to any one of claims 47 to 82, wherein when at 25°C for 1000 s -1 When measured at a shear rate of 100 mPa-s to 1000 mPa-s, the viscosity of the dynamic hydrogel encapsulating the acylated peptide was 100 mPa-s to 1000 mPa-s.

84. The method according to any one of claims 47 to 83, wherein, After administration to a subject, the composition delivers the acylated peptide to the subject for a treatment period of at least 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 28 days, 35 days, 40 days, 50 days, 60 days, 70 days, 80 days, 90 days, 100 days, 110 days, 120 days, 150 days, 180 days, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or 1 year.

85. The method according to any one of claims 47 to 84, wherein, After administration to a subject, the composition delivers the acylated peptide to the subject at rates of 0.25 mg / week to 25 mg / week, 0.5 mg / week to 20 mg / week, 0.5 mg / week to 2 mg / week, 0.5 mg / week to 1.5 mg / week, 1 mg / week to 2 mg / week, 2 mg / week to 5 mg / week, 5 mg / week to 15 mg / week, 5 mg / week to 10 mg / week, 10 mg / week to 20 mg / week, 10 mg / week to 15 mg / week, 12 mg / week to 25 mg / week, 12 mg / week to 15 mg / week, 15 mg / week to 25 mg / week, 15 mg / week to 20 mg / week, or 20 mg / week to 25 mg / week.

86. The method according to any one of claims 47 to 85, wherein, Upon administration to a subject, the composition produces an average concentration of the acylated peptide in the subject's serum of 50 ng / mL to 500 ng / mL, 100 ng / mL to 300 ng / mL, or 150 ng / mL to 250 ng / mL.

87. The method according to any one of claims 47 to 86, wherein, After administration to a subject, the composition produces C in the subject's serum at a concentration of less than or equal to 2000 ng / mL. max The acylated peptide.

88. The method according to any one of claims 47 to 87, wherein the composition is administered via subcutaneous injection.

89. The method according to any one of claims 47 to 88, wherein the disease or condition includes diabetes or diabetes-related conditions.

90. The method of claim 89, wherein the diabetes or diabetes-related condition includes prediabetes, type 1 diabetes, type 2 diabetes, hyperglycemia, or impaired glucose tolerance.

91. The method according to any one of claims 47 to 90, wherein the disease or condition includes one or more of obesity, overweight, eating disorders, or obstructive sleep apnea.

92. The method according to any one of claims 47 to 91, wherein the disease or condition includes one or more of the following: cardiovascular disease, liver disease, nervous system disease or neurodegenerative disease, inflammatory disease, kidney disease, bone disease, hormonal disease or gastrointestinal disease.

93. A composition for treating a disease or condition, said composition comprising: Dynamic hydrogel comprising a polymer and a plurality of nanoparticles, wherein the polymer is non-covalently crosslinked with the plurality of nanoparticles; and An incretin mimic, which is encapsulated in the dynamic hydrogel.

94. The composition of claim 93, wherein the incretin mimic comprises a glucagon-like peptide-1 receptor agonist (GLP-1 RA).

95. The composition of claim 94, wherein the GLP-1 RA comprises exenatide, exenatide-LAR, lixinatide, liraglutide, smegglutide, abiglutide, duraglutide, ipegglutide, enoglutide, ifenopeptide, crotonide, mascara, BI 45690, telpoglycinide, LY3493269, VK2735, CT-868, AMG133, or retaglutide.

96. The composition of claim 94, wherein the GLP-1 RA comprises liraglutide, smegglutide, enoglutide, cotodextrin, mastodextrin, telpogextrin, or retaglutide.

97. The composition according to any one of claims 94 to 96, wherein the GLP-1 RA comprises a peptide having a sequence having at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, or 99% sequence identity with any one of SEQ ID NO: 1 or SEQ ID NO:

2.

98. The composition according to any one of claims 93 to 97, wherein the incretin mimic binds only to the GLP-1 receptor.

99. The composition according to any one of claims 93 to 97, wherein the incretin mimic binds to a GLP-1 receptor and at least one additional receptor.

100. The composition of claim 99, wherein the at least one additional receptor comprises one or more of glucagon receptor or gastric inhibitory peptide (GIP) receptor.

101. The composition according to any one of claims 93 to 100, wherein the incretin mimic comprises an acylated peptide.

102. The composition of claim 101, wherein the acylated peptide comprises a lipophilic substituent.

103. The composition of claim 102, wherein the acylated peptide is encapsulated in the dynamic hydrogel via the interaction between the lipophilic substituent and the dynamic hydrogel.

104. The composition of claim 103, wherein the plurality of nanoparticles have a hydrophobic surface that interacts with the lipophilic substituents of the dynamic hydrogel.

105. The composition according to any one of claims 102 to 104, wherein the lipophilic substituent comprises an acyl group of a fatty acid.

106. The composition according to claim 105, wherein the fatty acid is a C4 fatty acid, C6 fatty acid, C8 fatty acid, C10 fatty acid, C12 fatty acid, C14 fatty acid, C15 fatty acid, C16 fatty acid, C17 fatty acid, C18 fatty acid, C20 fatty acid, C22 fatty acid, C24 fatty acid, C26 fatty acid, C28 fatty acid, C30 fatty acid, C32 fatty acid, C34 fatty acid, C36 fatty acid, or C38 fatty acid.

107. The composition according to any one of claims 102 to 106, further comprising a binding agent that interacts with the lipophilic substituent to physically encapsulate the acylated peptide in the dynamic hydrogel.

108. The composition of claim 107, wherein the binder comprises albumin.

109. The composition according to claim 107 or 108, wherein the binder is larger than the mesh size of the dynamic hydrogel.

110. The composition according to any one of claims 101 to 109, wherein the acylated peptide is smaller than the pore size of the dynamic hydrogel.

111. The composition according to any one of claims 101 to 110, further comprising a dispersant that inhibits the aggregation of the acylated peptide.

112. The composition of claim 111, wherein the dispersant comprises one or more of a surfactant or an osmotic pressure regulator.

113. The composition of claim 112, wherein the dispersant comprises the surfactant, and the surfactant comprises one or more of polysorbate, sorbitol fatty acid ester, poloxamer, polyoxyethylene alkyl ether, alkyl sulfate or its salt, fatty acid, fatty acid alcohol or phospholipid.

114. The composition according to claim 112 or 113, wherein the dispersant comprises the osmotic pressure regulator, and the osmotic pressure regulator comprises one or more of propylene glycol, glycerol, or mannitol.

115. The composition according to any one of claims 112 to 114, wherein the concentration of the dispersant in the composition is 0.1 mg / mL to 5 mg / mL, 0.5 mg / mL to 1.5 mg / mL, 1 mg / mL to 2 mg / mL, 1 mg / mL to 50 mg / mL, 5 mg / mL to 40 mg / mL, 10 mg / mL to 30 mg / mL, or 16 mg / mL to 26 mg / mL.

116. The composition according to any one of claims 93 to 100, wherein the incretin mimic comprises a peptide conjugated to a protein or protein fragment.

117. The composition according to any one of claims 93 to 100, wherein the incretin mimic comprises a peptide encapsulated in microparticles.

118. The composition according to any one of claims 93 to 117, wherein the composition comprises 1 mg to 250 mg of the incretin mimic.

119. The composition according to any one of claims 93 to 118, wherein the concentration of the incretin mimic in the composition is from 1 mg / mL to 200 mg / mL.

120. The composition according to any one of claims 93 to 119, wherein the polymer comprises a hydrophobically modified polysaccharide.

121. The composition of claim 120, wherein the hydrophobically modified polysaccharide comprises a hydrophobically modified cellulose derivative.

122. The composition according to claim 121, wherein the hydrophobically modified cellulose derivative is dodecyl modified hydroxypropyl methylcellulose (HPMC-C12).

123. The composition according to any one of claims 93 to 122, wherein the plurality of nanoparticles comprises a plurality of polymer nanoparticles.

124. The composition of claim 123, wherein the plurality of polymer nanoparticles are amphiphilic.

125. The composition according to claim 123 or 124, wherein the plurality of polymer nanoparticles are poly(ethylene glycol)-block-poly(lactic acid) (PEG-PLA) nanoparticles.

126. The composition according to any one of claims 93 to 125, wherein the concentration of the polymer in the dynamic hydrogel is 0.5 wt% to 5 wt%, 0.5 wt% to 4 wt%, 0.5 wt% to 3 wt%, 0.5 wt% to 2 wt%, 0.5 wt% to 1 wt%, 1 wt% to 5 wt%, 1 wt% to 4 wt%, 1 wt% to 3 wt%, 1 wt% to 2 wt%, 2 wt% to 5 wt%, 2 wt% to 4 wt%, 2 wt% to 3 wt%, 3 wt% to 5 wt%, 3 wt% to 4 wt%, or 4 wt% to 5 wt%.

127. The composition according to any one of claims 93 to 126, wherein the concentration of the nanoparticles in the dynamic hydrogel is 1 wt% to 12 wt%, 1 wt% to 10 wt%, 1 wt% to 8 wt%, 1 wt% to 5 wt%, 1 wt% to 3 wt%, 3 wt% to 12 wt%, 3 wt% to 10 wt%, 3 wt% to 8 wt%, 3 wt% to 5 wt%, 5 wt% to 12 wt%, 5 wt% to 10 wt%, 5 wt% to 8 wt%, 8 wt% to 12 wt%, 8 wt% to 10 wt%, or 10 wt% to 12 wt%.

128. The composition according to any one of claims 93 to 127, wherein the storage modulus of the dynamic hydrogel encapsulating the incretin mimic is 10 Pa to 1000 Pa or 50 Pa to 500 Pa when measured at 25°C at an angular frequency of 0.1 rad / s to 100 rad / s in the linear viscoelastic region of the dynamic hydrogel.

129. The composition according to any one of claims 93 to 128, wherein the yield stress of the dynamic hydrogel encapsulating the incretin mimic is 1 Pa to 500 Pa or 20 Pa to 200 Pa when measured at 25°C.

130. The composition according to any one of claims 93 to 129, wherein when at 25°C for 1000 s -1 When measured at a shear rate of 100 mPa-s to 1000 mPa-s, the viscosity of the dynamic hydrogel encapsulating the incretin mimic was 100 mPa-s to 1000 mPa-s.

131. The composition according to any one of claims 93 to 130, wherein, After administration to a subject, the composition delivers the incretin mimic to the subject for a treatment period of at least 7, 8, 9, 10, 11, 12, 13, 14, 28, 35, 40, 50, 60, 70, 80, 90, 100, 110, 120, 150, 180 days, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or 1 year.

132. The composition according to any one of claims 93 to 131, wherein, After administration to the subject, the composition delivers the incretin mimic to the subject at rates of 0.25 mg / week to 25 mg / week, 0.5 mg / week to 20 mg / week, 0.5 mg / week to 2 mg / week, 0.5 mg / week to 1.5 mg / week, 1 mg / week to 2 mg / week, 2 mg / week to 5 mg / week, 5 mg / week to 15 mg / week, 5 mg / week to 10 mg / week, 10 mg / week to 20 mg / week, 10 mg / week to 15 mg / week, 12 mg / week to 25 mg / week, 12 mg / week to 15 mg / week, 15 mg / week to 25 mg / week, 15 mg / week to 20 mg / week, or 20 mg / week to 25 mg / week.

133. The composition according to any one of claims 93 to 132, wherein, Upon administration to a subject, the composition produces an average concentration of the incretin mimic in the subject's serum of 50 ng / mL to 500 ng / mL, 100 ng / mL to 300 ng / mL, or 150 ng / mL to 250 ng / mL.

134. The composition according to any one of claims 93 to 133, wherein, After administration to a subject, the composition produces C in the subject's serum at a concentration of less than or equal to 2000 ng / mL. max The aforementioned incretin mimicry.

135. The composition according to any one of claims 93 to 134, wherein the composition is configured for administration via subcutaneous injection.

136. The composition according to any one of claims 93 to 135, wherein the disease or condition includes diabetes or diabetes-related conditions.

137. The composition of claim 136, wherein the diabetes or diabetes-related condition includes prediabetes, type 1 diabetes, type 2 diabetes, hyperglycemia, or impaired glucose tolerance.

138. The composition according to any one of claims 93 to 137, wherein the disease or condition includes one or more of obesity, overweight, eating disorders, or obstructive sleep apnea.

139. The composition according to any one of claims 93 to 138, wherein the disease or condition includes one or more of the following: cardiovascular disease, liver disease, nervous system disease or neurodegenerative disease, inflammatory disease, kidney disease, bone disease, hormonal disease or gastrointestinal disease.

140. A method for treating a disease or condition, said method comprising: The composition is applied to an object, wherein the composition comprises: Dynamic hydrogel comprising a polymer and a plurality of nanoparticles, wherein the polymer is non-covalently crosslinked with the plurality of nanoparticles; and An incretin mimic, which is encapsulated in the dynamic hydrogel.

141. The method of claim 140, wherein the incretin mimic comprises a glucagon-like peptide-1 receptor agonist (GLP-1 RA).

142. The method of claim 141, wherein the GLP-1 RA comprises exenatide, exenatide-LAR, lixinatide, liraglutide, smegglutide, abiglutide, duraglutide, ipegglutide, enoglutide, ifenopeptide, crotonide, mascara, BI 45690, telpoglycinide, LY3493269, VK2735, CT-868, AMG133, or retaglutide.

143. The method of claim 141, wherein the GLP-1 RA comprises liraglutide, smegglutide, enoglutide, cotodextrin, mastodextrin, telpogextrin, or retaglutide.

144. The method according to any one of claims 140 to 143, wherein the GLP-1 RA comprises a peptide having a sequence having at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, or 99% sequence identity with any one of SEQ ID NO: 1 or SEQ ID NO:

2.

145. The method according to any one of claims 140 to 144, wherein the incretin mimic binds only to the GLP-1 receptor.

146. The method according to any one of claims 140 to 144, wherein the incretin mimic binds to a GLP-1 receptor and at least one additional receptor.

147. The method of claim 146, wherein the at least one additional receptor comprises one or more of glucagon receptor or gastric inhibitory peptide (GIP) receptor.

148. The method according to any one of claims 140 to 147, wherein the incretin mimic comprises an acylated peptide.

149. The method of claim 148, wherein the acylated peptide comprises a lipophilic substituent.

150. The method of claim 149, wherein the acylated peptide is encapsulated in the dynamic hydrogel via the interaction between the lipophilic substituent and the dynamic hydrogel.

151. The method of claim 150, wherein the plurality of nanoparticles have a hydrophobic surface that interacts with the lipophilic substituents of the dynamic hydrogel.

152. The method according to any one of claims 149 to 151, wherein the lipophilic substituent comprises an acyl group of a fatty acid.

153. The method according to claim 152, wherein the fatty acid is a C4 fatty acid, C6 fatty acid, C8 fatty acid, C10 fatty acid, C12 fatty acid, C14 fatty acid, C15 fatty acid, C16 fatty acid, C17 fatty acid, C18 fatty acid, C20 fatty acid, C22 fatty acid, C24 fatty acid, C26 fatty acid, C28 fatty acid, C30 fatty acid, C32 fatty acid, C34 fatty acid, C36 fatty acid or C38 fatty acid.

154. The method according to any one of claims 149 to 153, wherein the composition further comprises a binding agent that interacts with the lipophilic substituent to physically encapsulate the acylated peptide in the dynamic hydrogel.

155. The method of claim 154, wherein the binding agent comprises albumin.

156. The method according to claim 154 or 155, wherein the binder is larger than the mesh size of the dynamic hydrogel.

157. The method according to any one of claims 148 to 156, wherein the acylated peptide is smaller than the pore size of the dynamic hydrogel.

158. The method according to any one of claims 148 to 157, wherein the composition further comprises a dispersant that inhibits the aggregation of the acylated peptide.

159. The method of claim 158, wherein the dispersant comprises one or more of a surfactant or an osmotic pressure regulator.

160. The method of claim 159, wherein the dispersant comprises the surfactant, and the surfactant comprises one or more of polysorbate, sorbitol fatty acid ester, poloxamer, polyoxyethylene alkyl ether, alkyl sulfate or its salt, fatty acid, fatty acid alcohol or phospholipid.

161. The method according to claim 159 or 160, wherein the dispersant comprises the osmotic pressure regulator, and the osmotic pressure regulator comprises one or more of propylene glycol, glycerol, or mannitol.

162. The method according to any one of claims 159 to 161, wherein the concentration of the dispersant in the composition is 0.1 mg / mL to 5 mg / mL, 0.5 mg / mL to 1.5 mg / mL, 1 mg / mL to 2 mg / mL, 1 mg / mL to 50 mg / mL, 5 mg / mL to 40 mg / mL, 10 mg / mL to 30 mg / mL, or 16 mg / mL to 26 mg / mL.

163. The method according to any one of claims 140 to 147, wherein the incretin mimic comprises a peptide conjugated to a protein or protein fragment.

164. The method according to any one of claims 140 to 147, wherein the incretin mimic comprises a peptide encapsulated in microparticles.

165. The method according to any one of claims 140 to 164, wherein the composition comprises 1 mg to 250 mg of the incretin mimic.

166. The method according to any one of claims 140 to 165, wherein the concentration of the incretin mimic in the composition is from 1 mg / mL to 200 mg / mL.

167. The method according to any one of claims 140 to 166, wherein the polymer comprises a hydrophobically modified polysaccharide.

168. The method of claim 167, wherein the hydrophobically modified polysaccharide comprises a hydrophobically modified cellulose derivative.

169. The method according to claim 168, wherein the hydrophobically modified cellulose derivative is dodecyl modified hydroxypropyl methylcellulose (HPMC-C12).

170. The method according to any one of claims 140 to 169, wherein the plurality of nanoparticles comprises a plurality of polymer nanoparticles.

171. The method of claim 170, wherein the plurality of polymer nanoparticles are amphiphilic.

172. The method according to claim 170 or 171, wherein the plurality of polymer nanoparticles are poly(ethylene glycol)-block-poly(lactic acid) (PEG-PLA) nanoparticles.

173. The method according to any one of claims 140 to 172, wherein the concentration of the polymer in the dynamic hydrogel is 0.5 wt% to 5 wt%, 0.5 wt% to 4 wt%, 0.5 wt% to 3 wt%, 0.5 wt% to 2 wt%, 0.5 wt% to 1 wt%, 1 wt% to 5 wt%, 1 wt% to 4 wt%, 1 wt% to 3 wt%, 1 wt% to 2 wt%, 2 wt% to 5 wt%, 2 wt% to 4 wt%, 2 wt% to 3 wt%, 3 wt% to 5 wt%, 3 wt% to 4 wt%, or 4 wt% to 5 wt%.

174. The method according to any one of claims 140 to 173, wherein the concentration of the nanoparticles in the dynamic hydrogel is 1 wt% to 12 wt%, 1 wt% to 10 wt%, 1 wt% to 8 wt%, 1 wt% to 5 wt%, 1 wt% to 3 wt%, 3 wt% to 12 wt%, 3 wt% to 10 wt%, 3 wt% to 8 wt%, 3 wt% to 5 wt%, 5 wt% to 12 wt%, 5 wt% to 10 wt%, 5 wt% to 8 wt%, 8 wt% to 12 wt%, 8 wt% to 10 wt%, or 10 wt% to 12 wt%.

175. The method according to any one of claims 140 to 174, wherein the storage modulus of the dynamic hydrogel encapsulating the incretin mimic is 10 Pa to 1000 Pa or 50 Pa to 500 Pa when measured at 25°C at an angular frequency of 0.1 rad / s to 100 rad / s in the linear viscoelastic region of the dynamic hydrogel.

176. The method according to any one of claims 140 to 175, wherein the yield stress of the dynamic hydrogel encapsulating the incretin mimic is 1 Pa to 500 Pa or 20 Pa to 200 Pa when measured at 25°C.

177. The method according to any one of claims 140 to 176, wherein when at 25°C for 1000 s -1 When measured at a shear rate of 100 mPa-s to 1000 mPa-s, the viscosity of the dynamic hydrogel encapsulating the incretin mimic was 100 mPa-s to 1000 mPa-s.

178. The method according to any one of claims 140 to 177, wherein, After administration to a subject, the composition delivers the incretin mimic to the subject for a treatment period of at least 7, 8, 9, 10, 11, 12, 13, 14, 28, 35, 40, 50, 60, 70, 80, 90, 100, 110, 120, 150, 180 days, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or 1 year.

179. The method according to any one of claims 140 to 178, wherein, After administration to the subject, the composition delivers the incretin mimic to the subject at rates of 0.25 mg / week to 25 mg / week, 0.5 mg / week to 20 mg / week, 0.5 mg / week to 2 mg / week, 0.5 mg / week to 1.5 mg / week, 1 mg / week to 2 mg / week, 2 mg / week to 5 mg / week, 5 mg / week to 15 mg / week, 5 mg / week to 10 mg / week, 10 mg / week to 20 mg / week, 10 mg / week to 15 mg / week, 12 mg / week to 25 mg / week, 12 mg / week to 15 mg / week, 15 mg / week to 25 mg / week, 15 mg / week to 20 mg / week, or 20 mg / week to 25 mg / week.

180. The method according to any one of claims 140 to 179, wherein, Upon administration to a subject, the composition produces an average concentration of the incretin mimic in the subject's serum of 50 ng / mL to 500 ng / mL, 100 ng / mL to 300 ng / mL, or 150 ng / mL to 250 ng / mL.

181. The method according to any one of claims 140 to 180, wherein, After administration to a subject, the composition produces C in the subject's serum at a concentration of less than or equal to 2000 ng / mL. max The aforementioned incretin mimicry.

182. The method according to any one of claims 140 to 181, wherein the composition is administered via subcutaneous injection.

183. The method according to any one of claims 140 to 182, wherein the disease or condition includes diabetes or diabetes-related conditions.

184. The method of claim 183, wherein the diabetes or diabetes-related condition includes prediabetes, type 1 diabetes, type 2 diabetes, hyperglycemia, or impaired glucose tolerance.

185. The method according to any one of claims 140 to 184, wherein the disease or condition includes one or more of obesity, overweight, eating disorders, or obstructive sleep apnea.

186. The method according to any one of claims 140 to 185, wherein the disease or condition includes one or more of the following: cardiovascular disease, liver disease, nervous system disease or neurodegenerative disease, inflammatory disease, kidney disease, bone disease, hormonal disease or gastrointestinal disease.

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