Stabilization of peptides and proteins in implantable devices and formulations

Abstract

The present disclosure relates to the field of patient treatment using an implantable delivery device for the long-term release of a therapeutic agent. In particular, the present disclosure relates to a device, a formulation, and a method for stabilizing peptide and protein formulations.

Description

Technology Field Cross-reference regarding related applications

[0001] This application claims priority to U.S. provisional patent application No. 63 / 617,535 filed January 4, 2024, the contents of which are incorporated herein by reference in their entirety. Background Technology

[0002] Many subjects, including not only human subjects but also veterinary subjects, require long-term treatment with therapeutic agents. To improve compliance, many subjects can benefit from the compliance provided by implantable devices that release the desired therapeutic agent at a desired rate over an extended period. In some cases, stabilization of the therapeutic agent is required.

[0003] There is a need in the field to protect therapeutic polypeptides and / or proteins placed within an implantable device from endogenous proteases and peptidases. The present disclosure meets this need and also provides other advantages.

[0004] In one embodiment, the present disclosure provides a device for treating the condition of an object, and the device is A capsule configured for transplantation and having a storage; Therapeutic agent placed within the storage; and It comprises a nanoporous membrane having multiple pores that provide a diffusion pathway for the therapeutic agent outside the reservoir; The therapeutic agent is a peptide or protein, and The device further includes a protease inhibitor.

[0005] In one embodiment, the therapeutic agent is an incretin mimic.

[0006] In one embodiment, the therapeutic agent is exenatide or semaglutide.

[0007] In one embodiment, the therapeutic agent is exenatide, liraglutide, semaglutide, dulaglutide, efpeglenatide, tyrzepatide, albiglutide, lixisenatide, retatrutide, pramlintide, surbodutide, cotadutide, cagrisema (cagrilintide + semaglutide) or an analogue or derivative thereof.

[0008] In one embodiment, the protease inhibitor is provided in a sustained-release configuration.

[0009] In one embodiment, the sustained-release formulation is a low-solubility form of a protease inhibitor.

[0010] In one embodiment, the protease inhibitor is incorporated into a sustained-release matrix, a sustained-release capsule, or a microparticle.

[0011] In one embodiment, the device further includes a solvent.

[0012] In one embodiment, the present disclosure provides a formulation for treating a condition of an object, the formulation is included in a device, and the device is A capsule configured for transplantation and having a storage; Treatment placed within the storage; It comprises a nanoporous membrane having multiple pores that provide a diffusion pathway for the therapeutic agent outside the reservoir; The therapeutic agent is a peptide or protein, and The device further includes a protease inhibitor.

[0013] In one embodiment, the present disclosure provides a method for treating the condition of an object, and the method As a step of providing a device, the device A capsule configured for transplantation and having a storage; Treatment placed within the storage; It comprises a nanoporous membrane having multiple pores that provide a diffusion pathway for the therapeutic agent outside the reservoir; The therapeutic agent is a peptide or protein, and The device further comprises a protease inhibitor, step; and It includes the step of implanting the device.

[0014] These and other aspects, purposes, and embodiments will become more apparent when read together with the detailed description and drawings of the present disclosure that follow. Brief explanation of the drawing

[0015] FIG. 1a shows a diagram of a device according to the present disclosure having a single storage.

[0016] FIG. 1b shows a diagram of a device according to the present disclosure having two repositories. Specific details for implementing the invention

[0017] The present disclosure relates to the field of organ treatment of a subject using an implantable device that provides continuous delivery of a therapeutic agent.

[0018] Embodiments of the present disclosure include formulations and devices comprising one or more therapeutic agents, and optionally one or more stabilizers, release rate regulators, or other beneficial agents for the therapeutic agents.

[0019] In some embodiments, the stabilizer is a protease inhibitor.

[0020] In some embodiments, the protease inhibitor is provided in a sustained-release configuration.

[0021] Additionally, embodiments of the present disclosure include a method for treating a subject using the device and formulation of the present invention. definition

[0022] "Polypeptide" refers to a molecule having a backbone chain of two or more amino acid residues. Some polypeptides may have additional associated groups, such as metal ions in metalloproteins, organic small molecules as in heme proteins, or carbohydrate groups as in glycoproteins.

[0023] “Peptide” and “protein” refer to subgroups of polypeptides or oligopeptides. In this disclosure, the definitions of peptide and protein follow the practice of the U.S. Food and Drug Administration (FDA), which defines peptide as a polypeptide having 40 or fewer amino acid residues and protein as a polypeptide having more than 40 amino acid residues.

[0024] Incretin mimics refer to preparations that act like incretin hormones and include preparations that bind to one or more of the human GLP-1 receptor, human GIP receptor, and human glucagon receptor, as well as preparations that bind to the human amylin receptor. Some incretin mimics include inhibitors of dipeptidyl peptidase-4 (DPP-4), which effectively increase the circulating levels of GLP-1 type peptides.

[0025] Those skilled in the art will understand that incretin mimics include, for example, exenatide, liraglutide, semaglutide, dulaglutide, tyrzepatide, albiglutide, lixisenatide, and cotadutide. Dual receptor incretin mimics include, for example, tyrzepatide, triple receptor incretin mimics include retatrutide, and amylin receptor incretin mimics include cagrilintide and pramlintide, and include analogs and derivatives of any of these agents, such as therapeutic agents having at least 95%, or 90%, or at least 85%, or 80%, or at least 75%, or 70% amino acid homology with any of these agents.

[0026] "Formulation of a therapeutic agent" refers to the actual state in which the therapeutic agent is present in a product or an intermediate for the manufacture of a product, and includes the therapeutic agent, and optionally, any additional therapeutic agent used, any formulation excipient used, and any formulation solvent used.

[0027] A "membrane" refers to a permeable structure that allows the mass transport of molecules to occur from one side of the structure through the structure to the other.

[0028] "Porous membrane" refers to a membrane characterized by the presence of a two-phase system, wherein the membrane matrix material exhibits one phase, typically a continuous phase, which is infiltrated by open channels extending from one side of the membrane to the other, and is filled with a second phase, often a fluid phase, through which mass transport through the membrane can occur.

[0029] "Dense" or "non-porous membranes" refer to fluid-filled membranes without pores. In such membranes, mass transport can occur via a dissolution-diffusion mechanism, where the therapeutic agent dissolves in the membrane material itself and permeates the membrane by diffusing through it.

[0030] "Nanoporous membrane" and "nanopore membrane" are used interchangeably and refer to a porous membrane having a minimum diameter of less than 1,000 nanometers.

[0031] "Nanotube membrane" refers to a nanoporous membrane, where the pores are formed by an array of nanotubes.

[0032] "Titania nanotube membrane" refers to an array of titania nanotubes on a titanium substrate, wherein at least some of the titania nanotubes have both ends open and can allow diffusion from one side of the membrane to the other through the titania nanotubes.

[0033] The "molecular diameter" of a polymer refers to the diameter of the polymer's radius of rotation sphere, which is a physical measure of molecular size and is defined as twice the mass-weighted average distance from the center of the molecule to each mass element within the molecule.

[0034] "Stokes diameter" or "hydrodynamic diameter" refers to the dimensions of a molecule and its associated water molecules as they move through an aqueous solution, and is defined as the radius of an equivalent hard sphere diffusing at the same speed as the molecule being observed.

[0035] "Ion exchange resin" refers to a polymer containing acidic or basic groups, or a combination thereof, which is made insoluble, for example, by cross-linking, and is capable of exchanging anions or cations, or a combination thereof, with a surrounding medium.

[0036] As used in this disclosure, “fluid” and “fluid form” refer to states of fluidable substances, including but not limited to gases, solutions, suspensions, emulsions, colloids, dispersions, etc.

[0037] "Fluid contact" refers to an object being in contact with a fluid.

[0038] "Neutral pH" refers to a pH of about 6.0 to about 8.0 or about 6.5 to about 7.5.

[0039] "Protease inhibitors" refer to molecules that inhibit the action of proteases. An example of a protease inhibitor is a compound that disables the active site of a protease by covalently modifying it. Proteases fall into various categories, including serine proteases, cysteine ​​proteases, threonine proteases, aspartate proteases, glutamine proteases, and metalloproteases. Examples of protease inhibitors include, but are not limited to, aprotinin (Trasylol), gabexate mesylate, benzamidine, and phenylmethylsulfonyl fluoride (PMSF). Serine protease inhibitors (serpines) and small serine protease inhibitors (smapin) are peptides or polysaccharides that protect the payload by irreversibly inhibiting serine proteases.

[0040] As used herein, “protease” generally refers to endogenous enzymes naturally produced in the body and includes trypsin, chymotrypsin, elastase (all serine proteases), pepsin (aspartate protease), cathepsin (cysteine ​​protease), substrate metalloprotease (MMP) (metalloprotease), thrombin, plasmin, and kallikrein; these enzymes play a role in digestion, blood coagulation, immune responses, and tissue remodeling.

[0041] The "administration" of a therapeutic agent refers to the act of injecting, implanting, or otherwise depositing the therapeutic agent by transdermally accessing the subject's body, such as in administration by injection or implantation means. The therapeutic agent may be contained within a drug delivery device, such as the device of FIG. 1a or FIG. 1b. The therapeutic agent may be delivered by the implantable device when the implantable device is administered.

[0042] "Administering" a therapeutic agent refers to the act of injecting, implanting, or otherwise depositing the therapeutic agent by transdermally accessing the subject's body, such as by administration via injection or implantation means. The therapeutic agent may be contained within a drug delivery device.

[0043] As used herein, the term "implant" broadly refers to any type of implanted or implantable object or device. Drug delivery devices or implants according to the present disclosure are not limited to any specific shape. The final shape and size of an implant in the body may vary and may differ depending on the therapeutic purpose. device

[0044] As illustrated in FIG. 1a, the device of the present invention comprises a capsule (101) suitable for implantation, and the capsule has a reservoir (102) suitable for holding therapeutic agents and stabilizers, release rate regulators, or other beneficial agents. In some embodiments, more than one reservoir exists. The capsule may be made of any suitable biocompatible material. In some embodiments, the capsule is made of a medical metal such as titanium or stainless steel, or of a medical polymer material such as silicon, polyurethane, polyacrylate, polyolefin, polyester, polyamide, etc. In some embodiments, the capsule is made of a plurality of materials. In some embodiments of the present disclosure, the capsule is made of titanium.

[0045] In some embodiments, the capsule is made of a single piece of material. In some embodiments, the capsule is made of multiple pieces of material and is, for example, a capsule having a cap having a membrane as a pathway for the release of a therapeutic agent, having a reservoir for holding a therapeutic agent and a stabilizer, a release rate regulator, or other beneficial agent, the cap may be attached to the reservoir by any suitable means such as welding, bonding, press-fitting, or the use of screw fastening means, or any combination thereof.

[0046] The capsule may have any suitable size or shape. In some embodiments of the present disclosure, the capsule is cylindrical, facilitating implantation into the body by a tubular implantation device such as a needle or trocar.

[0047] The device of the present disclosure has at least one membrane attached to a capsule and in fluid contact with a reservoir as described, and the membrane provides a pathway for mass transport of a therapeutic agent contained within the reservoir out of the reservoir and into the body of a subject to whom the capsule is implanted. In the present disclosure, “attached to a capsule” refers to a component being in a fixed position relative to the capsule and being connected directly or indirectly to the capsule using any suitable means including welding, bonding, press-fitting and the use of threaded means, or any combination thereof. In the case of the membrane, as described in U.S. Patent No. 9,814,867 and illustrated in FIG. 1a, the nanotube membrane is part of a nanotube array (103), some of which are still attached to a grown titanium substrate (104), and the substrate may be attached to the capsule. At least some of the nanotubes are open on both sides, as in (105) of FIG. 1a, to enable mass transport of the therapeutic agent out of the reservoir. The membrane may be attached to the capsule such that the substrate side faces the reservoir or faces away from the reservoir.

[0048] In certain cases, the device has a reservoir capacity with a volume of about 10 microliters to about 10 milliliters. In certain cases, the reservoir has a volume of about 10 μL to about 100 μL, e.g., 10, 20, 25, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, or 100 μL. In other cases, the reservoir is about 25-60 μL or about 30-50 μL, or even about 30-45 μL. In other cases, the reservoir is about 100-500 microliters, 150-450 microliters, or about 150-350 microliters, e.g., about 250 microliters.

[0049] Further descriptions of the device of the present disclosure can be found in U.S. Patent Publications Nos. US20220008345 and US20210246271, and U.S. Patent No. 9,814,867 and U.S. Patent No. 9,770,412, which are incorporated herein by reference.

[0050] FIG. 1b is a diagram of a device according to the present disclosure having two storage units separated by an enclosure. Various identification numbers in FIG. 1b correspond to the same features of FIG. 1a, but with the letter b added. membrane

[0051] An embodiment of the present disclosure includes at least one membrane that provides a route for mass transport of a therapeutic agent out of the storage of the device of the present disclosure.

[0052] In the embodiments of the present disclosure, various types of membranes may be used.

[0053] The membranes of the present disclosure comprise dense and porous membranes; the porous membrane comprises a nanoporous membrane and a nanotube membrane.

[0054] Materials suitable for the membrane of the present disclosure include organic and inorganic materials, polymers, ceramics, metals, metal oxides, and combinations thereof. Materials suitable for the membrane include silicon, silica, titanium, and titania.

[0055] In some embodiments, the membrane is a nanoporous membrane. In some embodiments, the membrane is a nanotube membrane. In some embodiments, the membrane is a titania nanotube membrane. Embodiments of the present disclosure are particularly useful as sustained delivery devices for therapeutic agents in which the release of the therapeutic agent is controlled by the nanoporous membrane.

[0056] Some embodiments of the present disclosure include a titania nanotube membrane as described in U.S. Patent No. 9,814,867. The pore size of the membrane of the present disclosure may be controlled by a process as described in U.S. Patent No. 9,770,412. Generally, the average pore size of the membrane of the present disclosure may be 1 to 1,000 nanometers. In some embodiments, an average pore size greater than 1,000 nanometers may exist. In some embodiments, the average pore size is 1 to 5 nanometers. In some embodiments, the average pore size is 5 to 10 nanometers. In some embodiments, the average pore size is 10 to 50 nanometers. In some embodiments, the average pore size is 50 to 100 nanometers. In some embodiments, the average pore size is 100 to 1,000 nanometers. In some embodiments, a pore size of less than 1 nanometer may exist.

[0057] In some embodiments, the membrane pores have a diameter of the same order of size as the hydrodynamic diameter of a dissolved substance, such as a therapeutic agent, within the formulation. In some embodiments, the pores have a diameter smaller than the hydrodynamic diameter of the dissolved substance within the formulation. Due to the finite size of the pores, these membranes can act as size-blocking filters for the dissolved substance within the formulation of a drug delivery system.

[0058] In some embodiments, the membrane pores have a diameter ranging from 1 to 5 times the molecular diameter of the drug molecule diffusing through the aqueous phase, or 1, 2, 3, 4, or 5 times. In some embodiments, the membrane pores have a diameter as described in U.S. Patent No. 11,129,791. Under these conditions, it has been found that a drug release rate can be achieved that is not controlled by the concentration gradient between the reservoir and the environment in which the drug is released, as is seen in typical Fickian diffusion, and can approach a more constant release rate over time.

[0059] The membrane pores are in fluid contact with the therapeutic agent within the reservoir, allowing molecules of the therapeutic agent to diffuse into and out of the pores and into the environment surrounding the device. The profile of the release rate over time can be any desired profile. In some embodiments, the profile is a decay profile following normal Pycian diffusion out of the reservoir. In some embodiments, the release rate profile is a non-Pycian profile, such as a constant rate or an approximate constant rate profile. A constant rate profile is sometimes referred to as a zero-order release rate profile. Some embodiments have a rapid increase in the drug release rate at the beginning of the profile. Some embodiments have a slow increase in the release rate at the beginning of the release rate profile.

[0060] The implantable drug delivery system of the present disclosure may have one or more membranes. For example, the implantable drug delivery system may have one, two, three, four or more membranes. Membrane types include nanoporous membranes and non-porous membranes. Different nanoporous membranes may have the same or different pore diameters. When the implantable drug delivery system has more than one membrane having the same pore diameter, each membrane may provide a diffusion pathway for the therapeutic agent. Alternatively, the membranes may each have different pore diameters so that one or more of the membranes do not provide a diffusion pathway for the therapeutic agent. In some embodiments, when two membranes are present in the implantable drug delivery system, optionally only one membrane provides a diffusion pathway for the therapeutic agent.

[0061] Further descriptions of the membranes of the present disclosure can be found in U.S. Patents No. 9,814,867 and 9,770,412. remedy

[0062] Some embodiments of the present disclosure include low molecular weight therapeutics, sometimes referred to as “small molecule drugs.” Some embodiments of the present disclosure include high molecular weight therapeutics, such as peptides and proteins, carbohydrates and nucleic acids, and combinations thereof, such as glycoproteins. If the therapeutic is a nucleic acid, a nuclease inhibitor may be present in the device.

[0063] Some embodiments of the present disclosure include more than one type of therapeutic agent.

[0064] The therapeutic agent of the present disclosure may exist in any desired state, including fluid and solid forms.

[0065] Any suitable therapeutic agent may be included in the embodiments of the present disclosure. For example, the therapeutic agent may be a small molecule drug having a molecular weight of less than about 1000 g / mol, or less than about 750 g / mol, or less than about 500 g / mol.

[0066] In some embodiments of the present disclosure, the therapeutic agent is a peptide or a protein. In some embodiments of the device, the therapeutic agent is an incretin mimic. In certain aspects, the therapeutic agent is present in a reservoir.

[0067] Exenatide (natural, recombinant, and synthetic, also known as exendin-4) refers to the compound with CAS number 141758-74-9; semaglutide (recombinant and synthetic) refers to the compound with CAS number 910463-68-2; liraglutide refers to the compound with CAS number 204656-20-2; dulaglutide refers to the compound with CAS number 923950-08-7; efpeglenatide refers to the compound with CAS number 1296200-77-5; tyrzepatide refers to the compound with CAS number 2023788-19-2; albiglutide refers to the compound with CAS number 782500-75-8; and lixisenatide refers to the compound with CAS number 320367-13-3; Retatrutide refers to the compound of CAS number 2381089-83-2; Framlintide refers to the compound of CAS number 196078-30-5; Surbodutide refers to the compound of CAS number 2805997-46-8; Cotadutide refers to the compound of CAS number 1686108-82-6; and Cagrisema (cagrilintide + semaglutide) refers to a formulation having the compound of CAS number 910463-68-2 and the compound of CAS number 1415456-99-3. These compounds may also include alternative pharmaceutical salts that retain the amino acid sequence.

[0068] In some embodiments, the amount of polypeptide therapeutic agent present is about 10 μg to about 50 mg, e.g., 100 μg, 200 μg, 300 μg, 400 μg, 500 μg, 600 μg, 700 μg, 800 μg, 900 μg, 1 mg, 10 mg, 20 mg, 30 mg, 40 mg, or 50 mg or more. Formulation

[0069] The formulations of the present disclosure comprise at least one therapeutic agent disposed in a reservoir, and optionally one or more stabilizers, release rate regulators, or other beneficial agents. Beneficial agents include buffers, solubility regulators, surfactants, soluble high molecular weight and low molecular weight stabilizers, antioxidants, antimicrobial agents, etc. A list of potentially suitable inactive ingredients used in medicines currently marketed in the United States can be found on the U.S. Food and Drug Administration (FDA) website.

[0070] The therapeutic agents and stabilizers, release rate regulators, or other beneficial formulations of the present disclosure may be combined in any suitable combination for manufacturing the device of the present disclosure in any suitable state by any suitable means.

[0071] In some cases, the formulations of the present disclosure are solid formulations, such as crystallized or freeze-dried powders.

[0072] In some cases, the formulations of the present disclosure are fluid or liquid formulations, such as true solutions.

[0073] In some cases, the formulations of the present disclosure are mixed formulations such as suspensions and emulsions.

[0074] Any of the components of a formulation including not only therapeutic agents but also stabilizers, release rate regulators, or other beneficial agents may exist in any of the above states, such as a suspension of a stabilizer in a drug solution or a suspension of a drug in an emulsion of a release rate regulator.

[0075] The solvent of the present disclosure may be any of an aqueous, organic, or mixed aqueous-organic solvent.

[0076] A therapeutic agent, stabilizer, release rate controller, or other beneficial agent in a fluid or solid state may be combined with a therapeutic agent in a fluid or solid state. A therapeutic agent with a stabilizer, release rate controller, or other beneficial agent may be combined in a solid state in a first step and converted to a fluid state in a subsequent step. A therapeutic agent with a stabilizer, release rate controller, or other beneficial agent may be combined in a fluid state in a first step and converted to a solid state in a subsequent step. The combination of a therapeutic agent with a stabilizer, release rate controller, or other beneficial agent may be carried out by any suitable method including dry powder mixing and the preparation of a fluid mixture of the therapeutic agent with the stabilizer, release rate controller, or other beneficial agent. These options may be combined in any suitable combination and permutation.

[0077] Some embodiments of the present disclosure include therapeutic agents that require stabilization. In some embodiments, stabilization is provided by a pH adjuster.

[0078] The stabilization mechanisms provided by embodiments of the present disclosure include chemical or physical mechanisms, and combinations of both. See also U.S. Patent Publication No. US20210246271.

[0079] Some embodiments of the present disclosure include therapeutic agents requiring control of the release rate. In some embodiments, control of the release rate is provided by a pH regulator, see US20220008345. In some embodiments, control of the release rate is provided by a surfactant, see WO2023 / 235302.

[0080] Some embodiments of the present disclosure include a stabilizer for a therapeutic peptide or protein in the form of a protease inhibitor. Some embodiments of the present disclosure include more than one protease inhibitor, such as one, two, three or more protease inhibitors.

[0081] In some cases, protease inhibitors are pH regulators, such as acidic molecules that lower the pH of a formulation to a level where the related protease or proteases are inactive, or basic molecules that raise the pH for the same purpose. A formulation for controlling the pH of a formulation is disclosed in U.S. Patent Publication No. 20220008345.

[0082] In some cases, protease inhibitors are metal ion chelators. Many proteolytic enzymes, such as substrate metalloproteases, depend on metal ions for their biological activity. By chelating these ions, the activity of these proteases is inhibited. Examples of metal ion chelators include, but are not limited to, dimercaprol, penicillamine, trientine, deferasirox, deferifron, deferoxamine, succimer, 2,2'-bipyridyl, EDTA, and EGTA.

[0083] In some cases, protease inhibitors are surfactants. Surfactants can inhibit protease activity by causing the unfolding of the active natural stereostructure of the protease inhibitor. Formulations containing surfactants are disclosed in WO2023 / 235302.

[0084] In some cases, protease inhibitors are declustering agents or detergents. Declustering agents act similarly to surfactants in that they destroy the active natural stereochemistry of protease inhibitors. Examples of declustering agents include, but are not limited to, quaternary ammonium ions such as urea and guanidinium chloride.

[0085] In some cases, protease inhibitors are compounds specifically developed or under investigation to inhibit proteases in disease conditions such as HIV, hepatitis C, Covid 19, and various types of cancer. Examples of such protease inhibitors include, but are not limited to, atazanavir, darunavir, indinavir, lopinavir, ritonavir, saquinavir, elpinavir, amprenavir, fosamprenavir, tipranavir, darunavir, boceprevir, telaprevir, simeprevir, asunaprevir, glecaprevir, grazoprevir, paritaprevir, remdesivir, marimastat, and batimastat.

[0086] In some cases, proteases are molecules generally known for their protease inhibitory activity, such as aprotinin, bestatin, chymostatin, and pepstatin.

[0087] In some cases, protease inhibitors are anti-cathepsin-L, clofazimin, glycopeptide antibiotics (e.g., teicoplanin, dalbavancin, oritavancin or telavancin), rifampicin, chloroquine, interferon (e.g., interferon a, interferon b or interferon c), clenbuterol or heparin.

[0088] The type of protease inhibitor can be experimentally determined using in vivo studies. The action of proteases is often highly specific to a particular amino acid or amino acid sequence. When protease-degrading activity is detected upon implantation of the device and peptide or protein formulation of the present disclosure, the identity of the associated protease can be revealed through characterization of the fragment. For example, cathepsin L tends to cleave amino acid chains between the N-terminal side of a dibasic residue and the dibasic residue.

[0089] Alternatively, possible proteolytic enzymes can be identified by recovering the internal solution of the extracted device of the present disclosure and performing proteomic analysis.

[0090] Based on the detected proteases, a suitable protease inhibitor may be selected. If the activity of more than one protease is detected, more than one protease inhibitor may be selected.

[0091] In certain cases, the concentration of the protease inhibitor is about 1 nM to about 100 mM. In certain cases, the concentration of the protease inhibitor is about 1 nM to about 500 nM. In certain cases, the concentration of the protease inhibitor is about 1 μM to about 800 μM. In other specific cases, the effective concentration range of the protease inhibitor is about 1 μM to 500 mM, or about 1-10 μM, or about 1 μM to about 500 μM, or about 500 μM to about 100 mM, or about 1 mM to about 50 mM.

[0092] In some embodiments, the composition of the present disclosure comprises a beneficial substance, such as a therapeutic agent, and a protease inhibitor for protecting the beneficial substance. In some cases, the protease inhibitor is significantly smaller than the therapeutic agent, and the release rate of the therapeutic agent through the membrane of the device is significantly slower than the release rate of the protease inhibitor, resulting in the premature depletion of the protease inhibitor. In certain cases, the amount of protease inhibitor present is increased to ensure that a sufficient amount exists to protect the therapeutic polypeptide or protein. In certain cases, the therapeutic agent is depleted before the protease inhibitor is depleted.

[0093] In some cases, protease inhibitors provide long-term stability through sustained-release mechanisms. Mechanisms include, but are not limited to, erosion, polymer biodegradation, dissolution, enzymatic cleavage, hydrolysis, proteolytic cleavage, encapsulation, etc.

[0094] For example, in a device using a nanotube membrane, the release rate of a therapeutic agent from a reservoir can be controlled by matching the dimensions of the nanotube to the molecular or hydrodynamic dimensions of the therapeutic agent in such a way that a sustained-release release rate profile is achieved. However, if the therapeutic agent requires a protease inhibitor, the dimensions of the protease inhibitor molecule are hardly matched to the dimensions of the therapeutic agent molecule, resulting in mismatched release of the two components. In the case of protein or peptide therapeutic agents, the protease inhibitor is almost always significantly smaller than the therapeutic agent molecule, leading to rapid depletion of the protease inhibitor from the reservoir. Some embodiments of the present disclosure provide a device and a composition for overcoming the problem of mismatched release rates of the therapeutic agent molecule and the protease inhibitor from the device.

[0095] In some embodiments, the device comprises a composition comprising a beneficial substance, such as a therapeutic agent, and one or more protease inhibitors, wherein the protease inhibitors provide long-term stability through a sustained-release mechanism. Such sustained-release mechanisms may include, for example, a degradable polymeric form of the protease inhibitor, a low-solubility formulation of the protease inhibitor, a protease inhibitor formulated as a sustained-release micro or macro formulation, or a protease inhibitor formulated as a membrane-encapsulated formulation. The terms polymer and oligomer are often used with somewhat overlapping meanings and refer to molecules composed of multiple monomer building blocks or subunits. In this disclosure, polymer refers collectively to any molecule composed of three or more monomer subunits.

[0096] In a sustained-release matrix, the carrier material and the released agent are mixed together as a mixture, where control of the release rate is achieved by the dimensions and composition of the mixture.

[0097] In sustained-release capsules, the released formulation within the center is surrounded by a shell or film of a release rate-regulating material, where control of the release rate is achieved by the dimensions and composition of the shell or film.

[0098] Additionally, or alternatively, providing a protease inhibitor on a sustained-release carrier can facilitate incorporation into the storage during manufacturing, unlike, for example, incorporating a precisely measured amount of powder.

[0099] In some embodiments of the present disclosure, a sustained-release formulation of a protease inhibitor is present in a reservoir, wherein the protease inhibitor is incorporated into a sustained-release matrix. Sustained-release matrices are well known in the field of pharmaceutical dosage form development, and many suitable types of matrices are available, including polymer and non-polymer matrices, and biodegradable and biostable matrices. Suitable manufacturing methods include bulk mixing of the components, e.g., mixing in a polymer mixer or mixer-extruder, followed by a sizing or size reduction step such as extrusion or milling. The preparation of controlled-release matrices in particle form is often carried out using a spray dryer.

[0100] Most of the emission rates of components dispersed in a Western-type matrix are commonly represented by the Higuchi equation, M t = K * t 0.5 It decreases over time according to, where M t ε is the fraction of the component released at time t, and K is a constant depending on the physicochemical properties of the matrix and the component. Therefore, the fraction of the released drug plotted against the square root of time becomes a straight line for 60%–80% of the component release.

[0101] In some embodiments of the present disclosure, a sustained-release formulation of a protease inhibitor is present in a reservoir, wherein the protease inhibitor is incorporated into a sustained-release capsule. The encapsulation of pharmaceutical ingredients is well known in the field of pharmaceutical dosage form development, and many suitable types of encapsulation materials are available, including polymer and non-polymer encapsulation materials, as well as biodegradable and biostable encapsulation materials. The manufacture of controlled-release capsules in particulate form is often performed using a fan coater, a spray coater, or a fluid bed coater.

[0102] The release rate of an ingredient contained within an encapsulated dosage form is determined by the thickness and permeability (constant value) of the encapsulating material to the ingredient, and the concentration gradient of the ingredient across the encapsulating membrane. As long as a solid ingredient is present within the encapsulated dosage form, the concentration gradient is approximately constant, and therefore the release rate over time is also constant.

[0103] Biodegradable encapsulation materials may include, but are not limited to, polyesters, polyphosphazenes, polyanhydrides, and polyorthoesters.

[0104] Biostable encapsulation materials may include, but are not limited to, polyolefins, polyvinyl, polyethers, polyurethanes, polyamides, polycarbonates, and silicones.

[0105] Non-polymer encapsulation materials may include, but are not limited to, waxes such as beeswax, microcrystalline wax, and paraffin wax.

[0106] The Western-type configuration can be in a microscopic or macroscopic form. In the microscopic form, the matrix can exist in the form of microparticles or nanoparticles. In the macroscopic form, the matrix can exist in any desired configuration, such as spheres, rods, strips, or configurations of irregular shapes.

[0107] A more extensive description of sustained-release technology applicable to the embodiments of the present disclosure is disclosed in U.S. Patent Publication No. 20160220496, which is incorporated herein by reference. A person skilled in the art of developing pharmaceutical dosage forms will be able to apply such technology without excessive experimentation.

[0108] In certain cases, the formulation is about 50 mg to about 400 mg, or about 100 to 300 mg, or about 150 to 250 mg. Treatment methods

[0109] Some embodiments of the present disclosure provide a method for treating a disease or condition in a subject using the device and formulation of the present disclosure. Some embodiments of the present disclosure provide a method for managing body weight. Some embodiments of the present disclosure provide a method for managing type 2 diabetes. Subjects include human and veterinary subjects.

[0110] The method comprises the steps of providing a device of the present disclosure comprising a therapeutic agent and, optionally, one or more stabilizers, release rate regulators, or other beneficial agents, and implanting the device into a subject to treat a disease or condition. Implantation may be performed by any means known to those skilled in the art, for example, by using a hollow needle or a trocar to insert the device subcutaneously.

[0111] In some embodiments, the present disclosure relates to a method comprising the steps of providing a weight management device suitable for implantation and implanting the device into a subject, wherein the step of providing the device is repeated at a frequency of less than once a week.

[0112] In some aspects, the device is implanted at a frequency of once a week to once a month, once a month to once every three months, once every three months to once every six months, once every six months to once every 12 months, and once every 12 months to once every 24 months.

[0112] For the purposes of the present disclosure, the frequency limits described in the present disclosure are included in the indicated periods, for example, once a month to once every three months includes a frequency of exactly one month to exactly three months.

[0113] Some embodiments of the present disclosure are methods for managing body weight in an object, Steps for providing weight management products, Step of administering a weight management agent to a subject, and A method is provided that includes the step of administering a weight management agent to a subject at a frequency of less than once a week.

[0114] In some aspects, the weight management agent is administered at one of the following frequencies: once a week to once a month, once a month to once every three months, once every three months to once every six months, once every six months to once every 12 months, and once every 12 months to once every 24 months.

[0115] The following patent documents are incorporated herein by reference: U.S. Patent Application Publications Nos. US 20220008345 and US 20210246271, and U.S. Patent Nos. 9,814,867, 9,770,412, 10,045,943, 10,479,868, 11,129,791 and WO2023 / 235302, the whole of which is incorporated herein by reference. Examples

[0116] In the following predictive example, the device used comprises a titanium capsule with a length of approximately 25 mm and a diameter of 2.25 mm. A titanium substrate having a titanium oxide nanoporous membrane is welded to one end of the device. The nanoporous membrane has a diameter of 0.3 mm and consists of approximately 6,000,000 nanopores. The average diameter of the nanopores at the end of the substrate is approximately 20 nm. A silicon diaphragm is located at the other end of the device.

[0117] As further described below, the sustained-release formulation of the manufactured protease inhibitor is incorporated into a reservoir before the membrane and septum are finally attached to the device.

[0118] The formulation solvent is filled into the device according to the methods of U.S. Patent Publication No. 20220008345 and U.S. Patent Publication No. 2021024627. Briefly, the formulation solution is loaded into a filling device having a hollow needle for penetrating a diaphragm. A vacuum is applied to the membrane of the device to lower the pressure inside the reservoir, and the formulation passes through the diaphragm via the needle and is injected into the reservoir.

[0119] The composition of the formulation solvent varies depending on the therapeutic agent being considered for the device. A typical formulation will be based on sterile water for injection containing 0.154 mM NaCl as an isotonic agent. An additional protease inhibitor may be required and may be provided by embodiments of the present disclosure.

[0120] The durability of the presence of a protease inhibitor within the device of the present disclosure can be tested by measuring the release of the protease inhibitor from the device in vitro. The in vitro test is performed by immersing the device in 3 ml of 26 mM bis-tris buffer, pH 7.4, 154 mM NaCl on a shaking plate at 37°C and measuring the amount of protease inhibitor released at regular intervals by reverse-phase HPLC. Example 1.

[0121] Marimastert, a protease inhibitor with anti-substrate metalloproteinase activity, and n-butyl methacrylate as a matrix polymer are co-dissolved in acetone in a ratio of 1-3. The total solid content of the solution is 10%. The solution is spray-dried into microparticles in a Buchi Mini Spray Dryer B-290.

[0122] The manufacturing and testing of the device are performed as described above.

[0123] Briefly, 10 milligrams of microparticles are loaded into a titanium reservoir, and the formulation is filled into the device as described. In vitro release rate tests are performed on a shaking plate at 37° in 26 mM bis-tris buffer, pH 7.4, and 154 mM NaCl. Sampling of the release rate solution and testing for marimastat are performed weekly to demonstrate the release of marimastat for at least two months. The release of marimastat demonstrates the presence of an active protease inhibitor within the device, thereby creating a protective environment for peptides and proteins against substrate metalloproteinases.

[0124] Calpeptin is an inhibitor of cathepsin L, a ubiquitous protease. Calpeptin release strips are prepared by mixing 1 part calpeptin with 2 parts of the silicon precursor MED-4830 Binary Silicon until a visually uniform paste is achieved. The paste is spread onto a glass sheet to form a film approximately 1 mm thick and cured at 85°C for 24 hours. The film is then cut into strips approximately 1 × 1 × 7 mm in size.

[0125] The manufacturing and testing of the device are performed as described above.

[0126] Briefly, a 10 mg calpeptin strip is loaded into a titanium reservoir, and the formulation is loaded into the device as described. In vitro release rate tests are performed on a shaking plate at 37° in 26 mM bis-tris buffer, pH 7.4, and 154 mM NaCl. Sampling of the release rate solution and testing for calpeptin are performed weekly to demonstrate the release of calpeptin for at least two months. The release of calpeptin demonstrates the presence of an active protease inhibitor within the formulation device, thereby creating a protective environment for peptides and proteins against cathepsin-L.

[0127] The embodiments and examples described herein are provided for illustrative purposes only, and various modifications or changes will be suggested to those skilled in the art in light of this, which are understood to be included within the spirit and scope of this application and the scope of the appended claims. All publications, patents, and patent applications cited herein are incorporated herein by reference in their entirety for all purposes.

Claims

Claim 1 A device for treating the condition of a subject, wherein the device comprises a capsule configured for implantation and having a reservoir, a therapeutic agent disposed within the reservoir, and a nanoporous membrane having a plurality of pores providing a diffusion path for the therapeutic agent outside the reservoir; wherein the therapeutic agent is a peptide or protein, and the device further comprises a protease inhibitor. Claim 2 A device in which the therapeutic agent in paragraph 1 is an incretin mimic. Claim 3 A device according to claim 1, wherein the therapeutic agent is a member selected from the group consisting of exenatide, liraglutide, semaglutide, dulaglutide, efpeglenatide, tirzepatide, albiglutide, lixisenatide, retatrutide, pramlintide, survodutide, cotadutide, cagrisema, and analogs or derivatives thereof. Claim 4 A device according to paragraph 2, wherein the incretin mimic is exenatide or semaglutide. Claim 5 A device according to any one of claims 1 to 4, wherein the protease inhibitor is provided in a sustained-release configuration. Claim 6 In paragraph 5, a device in which the sustained-release composition is a low-solubility form of a protease inhibitor. Claim 7 A device according to claim 5, wherein a protease inhibitor is incorporated into a sustained-release matrix or a sustained-release capsule. Claim 8 A device according to any one of claims 1 to 7, wherein the device further comprises a solvent. Claim 9 A formulation for treating the condition of a subject, wherein the formulation is included in a device, the device comprises a capsule configured for implantation and having a reservoir, a therapeutic agent disposed within the reservoir, and a nanoporous membrane having a plurality of pores providing a diffusion pathway for the therapeutic agent outside the reservoir; wherein the therapeutic agent is a peptide or a protein, and the device further comprises a protease inhibitor. Claim 10 In paragraph 9, a formulation in which the therapeutic agent is an incretin mimic. Claim 11 A formulation according to claim 9, wherein the therapeutic agent is a member selected from the group consisting of exenatide, liraglutide, semaglutide, dulaglutide, effeglenatide, tyrzepatide, albiglutide, lixisenatide, retatrutide, pramlintide, survodutide, cotadutide, cagrisema, and analogs or derivatives thereof. Claim 12 A device according to claim 10, wherein the incretin mimic is exenatide or semaglutide. Claim 13 A formulation according to any one of claims 9 to 12, wherein the protease inhibitor is provided in a sustained-release configuration. Claim 14 In paragraph 13, a formulation in which the sustained-release composition is a low-solubility form of a protease inhibitor. Claim 15 In paragraph 13, a formulation in which a protease inhibitor is incorporated into a sustained-release matrix or a sustained-release capsule. Claim 16 A formulation according to any one of claims 9 to 15, wherein the device further comprises a solvent. Claim 17 A method for treating the condition of a subject, the method comprising the step of providing a device, wherein the device comprises a capsule configured for implantation and having a reservoir, a therapeutic agent disposed within the reservoir, and a nanoporous membrane having a plurality of pores providing a diffusion path for the therapeutic agent outside the reservoir, wherein the therapeutic agent is a peptide or protein, and the device further comprises a protease inhibitor; and the step of implanting the device. Claim 18 In paragraph 17, a method in which the therapeutic agent is an incretin mimic. Claim 19 In paragraph 18, the method wherein the incretin mimic is exenatide or semaglutide. Claim 20 A method according to any one of claims 17 to 19, wherein the protease inhibitor is provided in a sustained-release configuration. Claim 21 In paragraph 20, the method wherein the sustained-release composition is a low-solubility form of a protease inhibitor. Claim 22 In paragraph 20, a method in which a protease inhibitor is incorporated into a sustained-release matrix or a sustained-release capsule. Claim 23 A method according to any one of claims 17 to 22, wherein the device further comprises a solvent. Claim 24 A method according to any one of claims 17 to 23, wherein the step of providing a weight management device suitable for implantation and the step of implanting said device into a subject are repeated at a frequency of less than once a week. Claim 25 A method according to claim 24, wherein the device is implanted at a frequency of one of once a week to once a month, once a month to once every three months, once every three months to once every six months, once every six months to once every twelve months, and once every twelve months to once every twenty-four months. Claim 26 A method for managing weight in a subject, comprising the steps of providing a weight management agent, administering the weight management agent to the subject, and repeating the step of administering the weight management agent to the subject at a frequency of less than once a week. Claim 27 A method according to claim 26, wherein a weight management agent is administered at one of the following frequencies: once a week to once a month, once a month to once every three months, once every three months to once every six months, once every six months to once every twelve months, and once every twelve months to once every twenty-four months.

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