Stabilization of peptides and proteins in implantable devices and formulations

Abstract

The disclosure pertains to the field of treatment of patients with implantable delivery devices for long-term release of therapeutic agents. In particular, the disclosure pertains to devices, formulations and methods to stabilize peptide and protein formulations.

Description

STABILIZATION OF PEPTIDES AND PROTEINS IN IMPLANTABLE DEVICES AND FORMULATIONSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to US Provisional Patent Application No. 63 / 617,535, filed January 4, 2024, the contents of which are hereby incorporated by reference in their entirety.BACKGROUND

[0002] Many subjects, human as well as veterinary, are in need of long-term treatment with therapeutic agents. In order to improve adherence, many subjects would benefit from the compliance provided by an implantable device releasing a desired therapeutic agent at a desired rate for an extended period of time. In some instances, stabilization of the therapeutic agents is required.

[0003] There is a need in the art for protecting therapeutic polypeptides and or proteins disposed within implantable devices from endogenous proteases and peptidases. The current disclosure satisfies this need and offers other advantages as well.BRIEF DESCRIPTION OF THE DISCLOSURE

[0004] In one embodiment, the disclosure provides a device for treating a condition in a subject, the device comprising: a capsule configured for implantation and having a reservoir; a therapeutic agent disposed within the reservoir; and a nanoporous membrane with a plurality of pores providing a diffusion path for the therapeutic agent out of the reservoir; wherein the therapeutic agent is a peptide or protein, the device further comprising a protease inhibitor.

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

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

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

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

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

[0010] In one embodiment, the protease inhibitor in incorporated in an extended-release matrix, extended-release capsule or microparticle.

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

[0012] In one embodiment, the disclosure provides a formulation for treating a condition in a subject, the formulation contained in a device, the device comprising: a capsule configured for implantation and having a reservoir; a therapeutic agent disposed within the reservoir; a nanoporous membrane with a plurality of pores providing a diffusion path for the therapeutic agent out of the reservoir; and wherein the therapeutic agent is a peptide or protein, the device further comprising a protease inhibitor.

[0013] In one embodiment the disclosure provides a method for treating a condition in a subject, the method comprising: providing a device, the device comprising: a capsule configured for implantation and having a reservoir; a therapeutic agent disposed within the reservoir; a nanoporous membrane with a plurality of pores providing a diffusion path for the therapeutic agent out of the reservoir; wherein the therapeutic agent is a peptide or protein,the device further comprising a protease inhibitor, and implanting the device.

[0014] These and other aspects, objects and embodiments will become more apparent when read with the detailed description of the disclosure and figures that follow.BRIEF DESCRIPTION OF THE DRAWINGS

[0015] FIG 1 A represents a diagram of a device according to the disclosure with one reservoir.

[0016] FIG IB represents a diagram of a device according to the disclosure with two reservoirs.DETAILED DESCRIPTION OF THE DISCLOSURE

[0017] The disclosure pertains to the field of long-term treatment of subjects with implantable devices providing a sustained delivery of therapeutic agents.

[0018] Embodiments of the disclosure include devices and formulations including one or more therapeutic agents and, optionally, one or more stabilizing, release rate modifying, or otherwise beneficial agents for the therapeutic agent.

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

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

[0021] Additionally, embodiments of the disclosure include methods of treatment of a subject with devices and formulations of the invention.DEFINITIONS

[0022] “Polypeptides” refers to molecules with a backbone chain of 2 or more amino acid residues. Some polypeptides may have additional associated groups, such as metal ions in metalloproteins, small organic molecules such as in heme proteins, or carbohydrate groups such as in glycoproteins.

[0023] “Peptides” and “Proteins” refers to subgroups of polypeptides or oligopeptides. In this disclosure the definition of peptides and proteins follows the practice of the United StatesFood and Drug Administration, the FDA, which defines peptides as polypeptides with up to 40 amino acid residues, and proteins as polypeptides with more than 40 amino acid residues.

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

[0025] A skilled person can appreciate that the incretin mimetics include, for example, exenatide, liraglutide, semaglutide, dulaglutide, tirzepatide, albiglutide, lixisenatide and cotadutide. Dual receptor incretin mimetics include, for example, tirzepatide, that triple receptor incretin mimetics include retatrutide, and that amylin receptor incretin mimetics include cagrilintide and pramlintide, as well as analogs and derivatives of any of these agents, such as therapeutic agents that have 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 a therapeutic agent is present in a product or in a product fabrication intermediate, and includes the therapeutic agent, plus, optionally, any used additional therapeutic agents, any used formulation excipients and any used formulation solvents.

[0027] “Membrane” refers to a permeable structure allowing mass transport of molecules from one side of the structure to the other through the structure.

[0028] “Porous membranes” refers to membranes characterized by the presence of a two- phase system, in which membrane matrix material represents one phase, typically a continuous phase, which is permeated by open channels extending from one side of the membrane to the other, and filled with a second phase, often a fluid phase, through which mass transport through the membrane can take place.

[0029] “Dense” or “non-porous membranes” refers to membranes without fluid filled pores. In such membranes mass transport may take place by a dissolution-diffusion mechanism, in which therapeutic agents permeate the membrane by dissolving in the membrane material itself, and diffusing through it.

[0030] “Nanoporous membrane” and “nanopore membrane” are used interchangeably, and refer to porous membranes in which the pores have a smallest diameter of less than 1000 nanometer.

[0031] “Nanotube membrane” refers to a nanoporous membrane, wherein pores are formed by an array of nanotubes.

[0032] “ Titania nanotube membrane” refers to an array of titania nanotubes on a titanium substrate where at least a portion of the titania nanotubes are open at both ends and capable of allowing diffusion from one side of the membrane to the other through the titania nanotubes.

[0033] ‘ Molecular diameter” of a polymer refers to the diameter of the sphere of gyration of the polymer, which is a physical measure of the size of a molecule, and is defined as two times the mass weighted average distance from the core of a molecule to each mass element in the molecule.

[0034] “ Stokes diameter” or “hydrodynamic diameter” refers to the dimension of a molecule plus its associated water molecules as it moves through an aqueous solution, and is defined as the radius of an equivalent hard sphere diffusing at the same rate as the molecule under observation.

[0035] “Ion exchange resin” refers to a polymer comprising acidic or basic groups, or a combination thereof, made insoluble, for instance by cross-linking, and capable of exchanging anions or cations, or a combination thereof, with a medium surrounding it.

[0036] “Fluid” and “fluid form” as used in this disclosure refers to flowable states of matter and includes, but is not limited to gases, solutions, suspensions, emulsions, colloids, dispersions and the like.

[0037] “Fluid contact” refers to an entity being in contact with a fluid.

[0038] “Neutral pH” refers to a pH between about 6.0 and about 8.0 or about 6.5 and about 7.5.

[0039] “Protease inhibitor” refers to a molecule that inhibits the action of proteases. One example of a protease inhibitor is a compound that covalently modifies the active site of the protease rendering it inoperative. There are various categories of proteases including serine proteases, cysteine proteases, threonine proteases, aspartate proteases, glutamine proteases and metalloproteases. Example of protease inhibitors include without limitation aprotinin(Trasylol), Gabexate mesylate, benzamidine and phenylmethyl sulfonyl fluoride (PMSF). Serine protease inhibitors (serpins) and small serine protease inhibitors (smapins) are peptides or polysaccharides that inhibit serine proteases irreversibly and thereby protect the payload.

[0040] “Protease” as used herein, is typically an endogenous enzyme, which is naturally produced within the body, and includes trypsin, chymotrypsin, elastase (all serine proteases), pepsin (aspartic protease), cathepsins (cysteine proteases), matrix metalloproteinases (MMPs) (metalloproteases), thrombin, plasmin, and kallikrein; these enzymes play roles in digestion, blood clotting, immune response, and tissue remodeling.

[0041] “Administration” of a therapeutic agent, as in administration by an injectable or implantable means, refers to the act of percutaneously accessing the body of a subject and injecting or implanting, or otherwise depositing, the therapeutic agent. The therapeutic agent may be contained within a drug delivery device such as the device of FIG. 1A or IB. The therapeutic agent may be delivered by an implantable device, once the implantable device is administered.

[0042] “Administering” a therapeutic agent, as in administering by an injectable or implantable means, refers to the act of percutaneously accessing the body of a subject and injecting or implanting, or otherwise depositing, the therapeutic agent. The therapeutic agent may be contained within a drug delivery device.

[0043] The term “implant”, as used herein, refers widely to any type of implanted or implantable object or device. The drug delivery device or implant according to the disclosure is not limited to any particular shape. The final shape and size of the implant in the body can vary and may depend on the purpose of the treatment.DEVICES

[0044] As illustrated in Fig. 1A, devices of the invention include a capsule 101 suitable for implantation, wherein the capsule has a reservoir 102 suitable for holding a therapeutic agent and a stabilizing, release rate modifying, or otherwise beneficial agent. In some embodiments, more than one reservoir is present. The capsule may be made of any suitable biocompatible material. In some embodiments the capsule is made of a medical grade metal, such as titanium or stainless steel, or of a medical grade polymeric material, such as silicone, polyurethane, polyacrylate, polyolefin, polyester, polyamide and the like. In someembodiments, the capsule is made of multiple materials. In some embodiments of the 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 materials, for instance a capsule having a reservoir for holding a therapeutic agent and a stabilizing, release rate modifying, or otherwise beneficial agent and having a cap holding a membrane as a pathway for release of the therapeutic agent, wherein the cap can be attached to the reservoir by any suitable means, such as welding, gluing, press fitting or using threaded means, or any combination of these.

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

[0047] Devices of the disclosure have at least one membrane, as described, attached to the capsule and in fluid contact with the reservoir, wherein the membrane provides a pathway for mass transport of a therapeutic agent included within the reservoir out of that reservoir and into the body of a subject into which the capsule has been implanted. In this disclosure “attached to the capsule” refers to a component being fixed in place with respect to the capsule, and connected to the capsule directly or indirectly, by using any suitable means, including by welding, gluing, press-fitting and by using threaded means, or by any combination of these. In the case of membranes as described in US Patent No. 9,814,867, and as illustrated in Fig. 1A, the nanotube membranes are part of an array of nanotubes 103, some of which are still attached to the titanium substrate 104 from which they were grown, and the substrate may be attached to the capsule. At least some of the nanotubes are open on both sides, 105 in Fig. 1 A, to allow for mass transport of a therapeutic agent out of the reservoir. The membranes may be attached to the capsule with the substrate side facing into the reservoir or facing away from the reservoir.

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

[0049] Further descriptions of devices of the disclosure may be found in US Patent Pub. Nos. US20220008345 and US20210246271, and US Patent No. 9,814,867 and US Patent No. 9,770,412, incorporated herein by reference.

[0050] FIG IB is a diagram of a device according to the disclosure with two reservoirs separated by an enclosure. The various identifying numbers in FIG. IB corresponding to the same features in FIG. 1 A but appended with the letter b.MEMBRANES

[0051] Embodiments of the disclosure include at least one membrane providing a pathway for mass transport of a therapeutic agent out of a reservoir of a device of the disclosure.

[0052] A wide variety of membranes can be used in embodiments of the present disclosure.

[0053] Membranes of the disclosure include dense and porous membranes; porous membranes include nanoporous membranes and nanotube membranes.

[0054] Suitable materials for membranes of the disclosure include organic and inorganic materials, polymers, ceramics, metals, metal oxides and combinations thereof. Suitable materials 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 disclosure are particularly useful as sustained delivery devices for therapeutic agents, in which the release of the agents is controlled by a nanoporous membrane.

[0056] Some embodiments of the disclosure comprise a titania nanotube membrane, such as described in US Patent No. 9,814,867. The pore size of membranes of the disclosure can be controlled by processes such as described in US Patent No. 9,770,412. Generally, average pore sizes of membranes of the disclosure may be between 1 and 1000 nanometer. In some embodiments, average pore sizes larger than 1000 nanometer may be present. In some embodiments the average pore size is from 1 to 5 nanometer. In some embodiments, the average pore size is from 5 to 10 nanometer. In some embodiments, the average pore size is from 10 to 50 nanometer. In some embodiments, the average pore size is from 50 to 100 nanometer. In some embodiments, the average pore size is from 100 to 1000 nanometer. In some embodiments, pore sizes of less than 1 nanometer may be present.

[0057] In some embodiments, the membrane pores have a diameter of the same order of magnitude as the hydrodynamic diameter of dissolved substances, such as a therapeutic agent in a formulation. In some embodiments, the pores have a diameter smaller than the hydrodynamic diameter of dissolved substances in a formulation. Because of the finite size of the pores, such membranes may act as a size cut-off filter for dissolved substances in the formulations of the drug delivery systems.

[0058] In some embodiments, the membrane pores have diameters in a range of 1-5 times or 1, 2, 3, 4, or 5 times the molecular diameter of the drug molecules diffusing through their aqueous phase. In some embodiments, the membrane pores have diameters, as described in, US Patent No. 11, 129,791. It has been shown that under those conditions drug release rates may be achieved that are not controlled by a concentration gradient between the reservoir and the environment into which the drug is released as would be seen in typical Fickian diffusion, and that may approach a more constant release rate over time.

[0059] The membrane pores are in fluid contact with the therapeutic agent in the reservoir, such that molecules of the therapeutic agent are able to diffuse into and out of the pores and into an environment surrounding the device. The profile of the release rate over time may be any desired profile. In some embodiments the profile is a declining profile, in accordance with regular Fickian diffusion out of the reservoir. In some embodiments the release rate profile is non-Fickian, like a constant rate or near-constant rate profile. Constant rate profiles are sometimes referred to as zero-order release rate profiles. Some embodiments, have a spike in drug release rate at early time points in the profile. Some embodiments have slow ramp up of release rates at early time points in the release rate profile.

[0060] The implantable drug delivery system of the present disclosure can have one or more membranes. For example, the implantable drug delivery system can have 1, 2, 3, 4, or more membranes. Membrane types include nanoporous and non-porous membranes. Different nanoporous membranes can have the same or different pore diameters. When the implantable drug delivery system has more than one membrane each with the same pore diameter, each membrane can provide a diffusion pathway for the therapeutic agent. Alternatively, the membranes can each have different pore diameters such that one or more of the membranes does not provide a diffusion pathway for the therapeutic agent. In some embodiments, when two membranes are present in the implantable drug delivery system, and optionally only one membrane provides a diffusion pathway for the therapeutic agent.

[0061] Further descriptions of membranes of the disclosure may be found in US Patent Nos. 9,814,867 and 9,770,412.THERAPEUTIC AGENTS

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

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

[0064] Therapeutic agents of the disclosure may be present in any desired state, including fluid and solid forms.

[0065] Any suitable therapeutic agent can be incorporated into embodiments of the disclosure. For example, the therapeutic agent can be a small molecule drug, such as one 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 disclosure the therapeutic agent is a peptide or protein. In some embodiments of the device the therapeutic agent is an incretin mimetic. In certain aspects, the therapeutic agent is present within the reservoir.

[0067] Exenatide (natural, recombinant and synthetic, also called exendin-4) refers to the compound with CAS No. 141758-74-9; Semaglutide (recombinant and synthetic) refers to the compound with CAS No. 910463-68-2; Liraglutide refers to the compound with CAS No.204656-20-2; Dulaglutide refers to the compound with CAS No. 923950-08-7; Efpeglenatide refers to the compound with CAS No. 1296200-77-5; Tirzepatide refers to the compound with CAS No. 2023788-19-2; Albiglutide refers to the compound with CAS No. 782500-75- 8; Lixisenatide refers to the compound with CAS No. 320367-13-3; Retatrutide refers to the compound with CAS No. 2381089-83-2; Pramlintide refers to the compound with CAS No. 196078-30-5; Survodutide refers to a compound with CAS No. 2805997-46-8; Cotadutide refers to a compound with CAS No. 1686108-82-6; and Cagrisema (cagrilintide+semaglutide) refers to a formulation having a compound with CAS No. 910463-68-2 and a compound with CAS No. 1415456-99-3. These compounds may also include alternative pharmaceutical salts, which maintain their amino acid sequences.

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

[0069] Formulations of the disclosure include at least one therapeutic agent and, optionally, one or more stabilizing, release rate modifying, or otherwise beneficial agents, disposed within the reservoir. Beneficial agents include buffering agents, solubility modifiers, surfactants, soluble high and low molecular weight stabilizers, anti-oxidants, antimicrobials and the like. A list of potentially suitable inactive ingredients used in currently marketed pharmaceutical products in the US can be found on the website of the United States Food and Drug Administration (FDA).

[0070] Therapeutic agents and stabilizing, release rate modifying, or otherwise beneficial agents in this disclosure may be combined in any suitable combination in preparing devices of the disclosure, by any suitable means, and in any suitable state.

[0071] In some instances, formulations of the disclosure are solid formulations, such as crystallized or lyophilized powders.

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

[0073] In some instances, formulations of the disclosures are mixed formulations, such as suspensions and emulsions.

[0074] Any of the components of the formulation, therapeutic agents as well as stabilizing, release rate modifying or otherwise beneficial agents can be in any of the above states, such as a suspension of stabilizer in a solution of a drug, or a suspension of a drug in an emulsion of a release rate modifying agent.

[0075] Solvents of the disclosure may be any of aqueous, organic, or mixed aqueous- organic solvents.

[0076] Therapeutic agents, stabilizing, release rate modifying, or otherwise beneficial agents in a fluid or solid state may be combined with therapeutic agents in a fluid or solidstate. Stabilizing, release rate modifying, or otherwise beneficial agents and therapeutic agents may be combined in their solid states in a first step, and brought into a fluid state in a later step. Stabilizing, release rate modifying, or otherwise beneficial agents and therapeutic agents may be combined in fluid states in a first step, and brought into a solid state in a later step. Combination of the therapeutic agent and the stabilizing, release rate modifying, or otherwise beneficial agent may be performed by any suitable method, including dry powder mixing and preparing a fluid mixture of the therapeutic agent and the stabilizing, release rate modifying, or otherwise beneficial agent. These options may be combined in any suitable combination and permutation.

[0077] Some embodiments of the disclosure comprise a therapeutic agent in need of stabilization. In some embodiments, stabilization is provided by pH-controlling agents.

[0078] Stabilization mechanisms provided by embodiments of the disclosure include chemical or physical mechanisms, as well as combinations of both. See also US Patent Pub. No. US20210246271.

[0079] Some embodiments of the disclosure comprise a therapeutic agent in need of release rate modification. In some embodiments release rate modification is provided by pH- controlling agents, see US20220008345. In some embodiments release rate modification is provided by surfactants, see WO2023 / 235302.

[0080] Some embodiments of the disclosure comprise a stabilizer for a therapeutic peptide or protein in the form of a protease inhibitor. Some embodiments of the disclosure comprise more than one protease inhibitor, such as 1, 2, 3 or more protease inhibitors.

[0081] In some instances, the protease inhibitor is a pH modifier, such as an acidic molecule, lowering the pH of a formulation to a level where the relevant protease or proteases is or are inactive, or a basic molecule increasing the pH for the same purpose. Formulations to control the pH of a formulation have been disclosed in US Patent Pub. No. 20220008345.

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

[0083] In some instances, the protease inhibitor is a surfactant. Surfactants may inhibit protease activity by causing unfolding of the active native conformation of the protease inhibitor. Formulations with surfactants have been disclosed in WO2023 / 235302.

[0084] In some instances, the protease inhibitor is a declustering agent or detergent. Declustering agents work similarly to surfactants, in that they disrupt the active native conformation of the protease inhibitor. Examples of declustering agents include, but are not limited to, urea and quaternary ammonium ions, such as guanidinium chloride.

[0085] In some instances, the protease inhibitor is a compound specifically developed or under investigation to inhibit proteases in disease states, 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, elfinavir, amprenavir, fosamprenavir, tipranavir, darunavir, boceprevir, telaprevir and simeprevir, asunaprevir, glecaprevir, grazoprevir, paritaprevir and remdesivir, marimastat and batimastat.

[0086] In some instances, the protease is a molecule generally known for protease inhibitor activity, such as but not limited to aprotinin, bestatin and chymostatin and pepstatins.

[0087] In some instances, the protease inhibitor is anti-cathepsin-L, clofamizine, a glycopeptide antibiotic (such as teicoplanin, dalbavancin, oritavancin or telavancin) rifampicin, chloroquine, an interferon (such as interferon a, interferon b or interferon c), clenbuterol or heparin.

[0088] The type of protease inhibitor can be determined experimentally using in-vivo studies. The action of proteolytic enzymes is often highly specific for certain amino acids or amino acid sequences. If, upon implantation of a device and a peptide or protein formulation of the disclosure, proteolytic activity is detected, identification of the nature of the fragments may reveal the identity of the protease involved. For instance, cathepsin L tends to cleave amino acids chains at the N-terminal side of dibasic residues as well as between the dibasic residues.

[0089] Alternatively, inside solutions of explanted devices of the disclosure can be retrieved, and proteomic analysis can be performed to identify possible proteolytic enzymes.

[0090] Based on detected proteases, suitable protease inhibitors can be selected. In those cases where activity of more than one protease is detected, more than one protease inhibitor may be selected.

[0091] In certain instances, the protease inhibitor concentration is about 1 nM to about 100 mM. In certain instances, the protease inhibitor concentration is about 1 nM to about 500 nM. In certain instances, the protease inhibitor concentration is about 1 pM to about 800 pM. In certain other instances, the effective concentration range for the protease inhibitor is about 1 pM to 500 mM, or about 1-10 pM, or about 1 pM to about 500 pM, or about 500 pM to about 100 mM, or about 1 mM to about 50 mM.

[0092] In some embodiments, compositions of the disclosure comprise a beneficial substance, such as a therapeutic agent and a protease inhibitor for protection of the beneficial substance. In some instances, the protease inhibitors are significantly smaller than the therapeutic agents and the rate of release of the therapeutic agent through a membrane of a device is significantly slower than the rate of release of the protease inhibitor, leading to premature depletion of the protease inhibitor. In certain instances, the amount of protease inhibitor present is increased to ensure enough is present to protect the therapeutic polypeptide or protein. In certain instances, the therapeutic agent is depleted before the protease inhibitor is depleted.

[0093] In some instances, the protease inhibitor provides long term stability through an extended-release mechanism for the protease inhibitor. Mechanisms include, but are not limited to, erosion, biodegradation of a polymer, dissolution, enzymatic cleavage, hydrolysis, proteolytic cleavage, encapsulation and the like.

[0094] For instance, in devices using a nanotube membrane, the rate of release of a therapeutic agent from the reservoir may be controlled by matching the dimensions of the nanotubes to molecular dimensions or hydrodynamic dimensions of the therapeutic agent in such a manner that an extended-release rate profile is achieved. However, when the therapeutic agent requires a protease inhibitor, the dimensions of protease inhibitor molecules rarely match the dimensions of the therapeutic agent molecules, resulting in unmatched release of the two components. In the case of protein or peptide therapeutic agents, the protease inhibitors are almost always significantly smaller than the therapeutic agent molecules, leading to rapid depletion of the protease inhibitor from the reservoir. Some embodiments of the present disclosure provide devices and compositions to overcome the problem of unmatched release rates of therapeutic agent molecules and protease inhibitors from a device.

[0095] In some embodiments, the devices comprise a composition that includes a beneficial substance, such as a therapeutic agent, as well as one or more protease inhibitors, wherein the protease inhibitors provide long term stability through an extended-release mechanism. Such extended-release mechanisms may include, for instance, degradable polymeric forms of the protease inhibitor, formulations of low solubility forms of the protease inhibitor, protease inhibitors formulated into slow release micro- or macro formulations, or protease inhibitors formulated into a membrane-encapsulated formulation. The terms polymer and oligomer are often used in a somewhat overlapping sense, and refer to molecules composed of multiple monomeric building blocks or subunits. Polymer, in this disclosure, collectively refers to any molecule composed of more than three monomeric subunits.

[0096] In sustained release matrices, the carrier material and the released agent are mixed together into a mixture, wherein control over the release rate is achieved by the dimensions and composition of the mixture.

[0097] In sustained release capsules, the released agent in a core is surrounded by a shell or film of a release rate controlling material, wherein control over the release rate is achieved by the dimensions and composition of the shell or film.

[0098] Additionally, or alternatively, providing protease inhibitors in a sustained release carrier may facilitate their incorporation in the reservoir during manufacturing, as opposed to, for instance, incorporating a precisely weighed out quantity of a powder.

[0099] In some embodiments of the disclosure, an extended-release configuration of a protease inhibitor is present in a reservoir, wherein the protease inhibitor is incorporated in a sustained-release matrix. Sustained release matrices are well known in the art of pharmaceutical dosage form development, and many suitable types of matrices are available, including polymeric and non-polymeric matrices and biodegradable and bio-stable matrices. Suitable preparation methods include bulk mixing of ingredients, for instance in a polymer mixer or mixer-extruder, followed by a sizing or size reduction step, such as extrusion or milling. Preparation of controlled release matrices in particulate form is often performed with spray dryers.

[0100] A major portion of the release rate of ingredients dispersed in a sustained-release matrix often declines over time according to the Higuchi equation, Mt = K * t°5, where Mt is the fraction of the ingredient released at time t, and K is a constant depending on the matrix and the physicochemical properties of the ingredient. Hence the fraction of drug releasedplotted against the square root of time becomes a straight line for 60%-80% of the release of the ingredients.

[0101] In some embodiments of the disclosure, an extended-release configuration of a protease inhibitor is present in a reservoir, wherein the protease inhibitor is incorporated in a sustained release capsule. Encapsulation of pharmaceutical ingredients is well-known in the art of pharmaceutical dosage form development, and many suitable type of encapsulation materials are available, including polymeric and non-polymeric encapsulation materials, and biodegradable and bio-stable encapsulation materials. Preparation of controlled release capsules in particulate form is often performed with pan coaters, spray coaters or fluidized bed coaters.

[0102] The release rate of ingredients contained within an encapsulated dosage form are determined by the thickness and permeability of the encapsulation material for the ingredient (Constant values), and the concentration gradient of the ingredient over the encapsulating membrane. As long as solid ingredient is present inside the encapsulated dosage form the concentration gradient is approximately constant, and therefore so is the release rate over time.

[0103] Biodegradable encapsulation materials can include, but are not limited to, polyesters, poly-phosphazenes, poly-anhydrides, and poly-ortho esters.

[0104] Biostable encapsulation materials can include, but are not limited to, poly-olefines, poly-vinyls, poly-ethers, poly-urethanes, poly-amides poly-carbonates and silicones.

[0105] Non-polymeric encapsulation materials can include, but are not limited to, waxes, such as bees wax, microcrystalline wax and paraffin wax.

[0106] The extended-release configurations can be in microscopic or macroscopic form. In microscopic form the matrices can be present in microparticle or nanoparticle form. In macroscopic form the matrices can be present in any desired configuration, such as spheres, rods, strips, or irregular shaped configurations.

[0107] More extensive descriptions of extended-release technologies applicable to embodiments of the disclosure are disclosed in US Patent Pub. No. 20160220496, incorporated herein by reference. Those with ordinary skills in the art of pharmaceutical dosage form development will be able to apply such technologies without undue experimentation.

[0108] In certain instances, the formulation is about 50 mg to about 400 mg, or about 100- 300 mg, or about 150-250 mg.TREATMENT METHODS

[0109] Some embodiments of the disclosure provide methods of treating a disease or condition in subjects using devices and formulations of the disclosure. Some embodiments of the disclosure provide methods to manage body weight. Some embodiments of the disclosure provide methods to manage Type 2 Diabetes. Subjects include human and veterinary subjects.

[0110] The methods include providing a device of the disclosure including a therapeutic agent and, optionally, one or more stabilizing, release rate modifying, or otherwise beneficial agents and implanting the device in the subject, thereby treating the disease or condition. The implantation can be performed by any means known to one of skill in the art, for instance through subcutaneous insertion of the device using a hollow needle or trocar.[OHl] Some embodiments, the disclosure relates to methods of providing a device for weight management suitable for implantation and implanting the device in the subject, wherein the step of providing the device is repeated with a frequency of less frequent than once a week.

[0112] In some aspects, a device is implanted with a frequency that is one of: between once a week and once a month, and between once a month and once every 3 months, and between once every 3 months and once every 6 months, and between once every 6 months and once every 12 months, and between once every 12 months and once every 24 months.

[0113] For purposes of this disclosure, the limits of the frequencies described in the disclosure are included in the time periods indicated, such that, for instance, a frequency of between once a month and once every 3 months includes the frequency of exactly one month to exactly 3 months.

[0114] Some embodiments of the disclosure provide method of managing body weight in a subject, comprising: 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 with a frequency of less frequent than once a week.

[0115] In some aspects, the weight management agent is administered with a frequency that is one of between once a week and once a month, and between once a month and once every 3 months, and between once every 3 months and once every 6 months, and between once every 6 months and once every 12 months, and between once every 12 months and once every 24 months.

[0116] The following patent documents are hereby incorproarted by refence: US Patent Application Publication Nos. US 20220008345 and US 20210246271, and US Patents Nos. 9,814,867, 9,770,412, 10,045,943, 10,479,868, 11,129,791 and WO2023 / 235302, which are incorporated herein in their entireties by reference.EXAMPLES

[0117] In the prophetic examples below, the devices that are used include titanium capsules of approximately 25 mm length and 2.25 mm diameter. A titanium substrate with a titanium oxide nanoporous membrane is welded to one end of the device. The nanoporous membrane has a diameter of 0.3 mm and is composed of about 6,000,000 nanopores. The average diameter of the nanopores at the substrate end is approximately 20 nm. A silicone septum is located at the other end of the device.

[0118] Extended-release configurations of protease inhibitors, prepared as described further below, are incorporated into the reservoirs before final attachment of the membrane and septum on the device.

[0119] A formulation solvent is filled into the device as per methods in US Patent Pub No. 20220008345 and US Patent Pub No. 2021024627. Briefly, the formulation solution is loaded into a filler apparatus with a hollow needle to pierce the septum. A vacuum is applied to the membrane of the device to reduce the pressure inside the reservoir, and the formulation is injected through the septum into the reservoir through the needle.

[0120] The composition of the formulation solvent depends on the therapeutic agent being considered for the device. Typical formulations will be based on sterile water for injection with 0.154mM NaCl as a tonicity agent. Additional protease inhibitors may be required, and may be may be provided by embodiments of the disclosure.

[0121] The durability of the presence of protease inhibitors inside devices of the disclosure may be tested by measuring release of the protease inhibitors from the devices in vitro. Invitro testing is performed by submerging the devices in 3 ml of a 26 mM bis-tris buffer, pH 7.4, 154 mM NaCl on a shaker plate at 37°C and measuring the amounts of protease inhibitor released at regular intervals by reverse phase HPLC.EXAMPLE 1.

[0122] Marimastat, a protease inhibitor with anti-matrix-metalloproteinase activity, and n- butyl-methacrylate as a matrix polymer are co-dissolved in acetone in a 1-3 ratio. The total solid content of the solution is 10%. The solution is spray-dried into microparticles on a Buchi Mini Spray Dryer B-290.

[0123] Preparation and testing of the devices is performed as described above.

[0124] Briefly, 10 milligrams of microparticles are loaded into a titanium reservoir, after which the device is filled with formulation as described. In vitro release rate testing is performed in 26 mM bis-tris buffer, pH 7.4, 154 mM NaCl on a shaker plate at 37°. Sampling of release rate solution and testing for marimastat is performed on a weekly basis, and shows release of marimastat for at least 2 months. The release of marimastat demonstrates the presence of active protease inhibitor inside the device, thereby creating a protective environment for peptides and proteins against matrix metalloproteinases.

[0125] Calpeptin is an inhibitor of the ubiquitous protease Cathepsin L. Calpeptin releasing strips are manufactured by mixing 1 part calpeptin with 2 parts of silicone precursor MED- 4830 Binary Silicon until a visually uniform paste was achieved. The paste is drawn into a film of about 1 mm thick on a glass sheet, and cured for 24 hours minutes at 85°C. The film is then cut into strips of approximately 1x1x7mm.

[0126] Preparation and testing of the devices is performed as described above.

[0127] Briefly, 10 milligrams of calpeptin strips are loaded into a titanium reservoir, after which the device is filled with formulation as described. In vitro release rate testing is performed in 26 mM bis-tris buffer, pH 7.4, 154 mM NaCl on a shaker plate at 37°. Sampling of release rate solution and testing for calpeptin is performed on a weekly basis, and shows release of calpeptin for at least 2 months. The release of calpeptin demonstrates the presence of active protease inhibiter inside the formulation device, thereby creating a protective environment for peptides and proteins against cathepsin-L.

[0128] It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will besuggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.

Claims

WHAT IS CLAIMED IS:1 A device for treating a condition in a subject, the device comprising: a capsule configured for implantation and having a reservoir; a therapeutic agent disposed within the reservoir; and a nanoporous membrane with a plurality of pores providing a diffusion path for the therapeutic agent out of the reservoir; wherein the therapeutic agent is a peptide or protein, the device further comprising a protease inhibitor.

2. The device of claim 1, wherein therapeutic agent is an incretin mimetic.

3. The device of claim 1, wherein 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 an analog or derivative thereof.

4. The device of claim 2, wherein incretin mimetic is exenatide or semaglutide.

5. The device of any one of claims 1-4, wherein the protease inhibitor is provided in an extended-release configuration.

6. The device of claim 5, wherein the extended-release configuration is a low-solubility form of the protease inhibitor.

7. The device of claim 5, wherein the protease inhibitor in incorporated in an extended-release matrix or extended-release capsule.

8. The device of any one of claims 1-7, wherein the device further comprises a solvent.

9. A formulation for treating a condition in a subject, the formulation contained in a device, the device comprising: a capsule configured for implantation and having a reservoir;a therapeutic agent disposed within the reservoir; and a nanoporous membrane with a plurality of pores providing a diffusion path for the therapeutic agent out of the reservoir; wherein the therapeutic agent is a peptide or protein, the device further comprising a protease inhibitor.

10. The formulation of claim 9, wherein therapeutic agent is an incretin mimetic.

11. The formulation of claim 9, wherein 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 an analog or derivative thereof.

12. The device of claim 10, wherein incretin mimetic is exenatide or semaglutide.

13. The formulation of any one of claims 9-12, wherein the protease inhibitor is provided in an extended-release configuration.

14. The formulation of claim 13, wherein the extended-release configuration is a low-solubility form of the protease inhibitor.

15. The formulation of claim 13, wherein the protease inhibitor in incorporated in an extended-release matrix or extended-release capsule.

16. The formulation of any one of claims 9-15, wherein the device further comprises a solvent.

17. A method for treating a condition in a subject, the method comprising: providing a device, the device comprising: a capsule configured for implantation and having a reservoir; a therapeutic agent disposed within the reservoir; and a nanoporous membrane with a plurality of pores providing a diffusion path for the therapeutic agent out of the reservoir; wherein the therapeutic agent is a peptide or protein, the device further comprising a protease inhibitor, andimplanting the device.

18. The method of claim 17, wherein therapeutic agent is an incretin mimetic.

19. The method of claim 18, wherein incretin mimetic is exenatide or semaglutide.

20. The method of any one of claims 17-19, wherein the protease inhibitor is provided in an extended-release configuration.

21. The method of claim 20, wherein the extended-release configuration is a low-solubility form of the protease inhibitor.

22. The method of claim 20, wherein the protease inhibitor in incorporated in an extended-release matrix or extended-release capsule.

23. The method of any one of claims 17-22, wherein the device further comprises a solvent.

24. The method of any of claims 17-23, wherein the steps of providing a device for weight management suitable for implantation and implanting the device in the subject are repeated with a frequency of less frequent than once a week.

25. The method of claim 24, wherein a device is implanted with a frequency that is one of: between once a week and once a month, and between once a month and once every 3 months, and between once every 3 months and once every 6 months, and between once every 6 months and once every 12 months, and between once every 12 months and once every 24 months.

26. A method of managing body weight in a subject, comprising 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 with a frequency of less frequent than once a week.

27. The method of claim 26, wherein the weight management agent is administered with a frequency that is one of: between once a week and once a month, and between once a month and once every 3 months, and between once every 3 months and once every 6 months, and between once every 6 months and once every 12 months, and between once every 12 months and once every 24 months.

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