Novel pharmaceutical compositions containing glucagon-like peptide-1 receptor agonists

Abstract

A pharmaceutical formulation useful for treating metabolic disorders or conditions is provided, comprising a plurality of particles suspended in a carrier system, the particles (a) having an average diameter based on weight, number, or volume of about 10 nm to about 700 μm, and (b) a solid core comprising at least one glucagon-like peptide-1 receptor agonist or a pharmaceutically acceptable salt thereof, at least partially coated with a coating of an inorganic material comprising a mixture of (i) zinc oxide and (ii) one or more other metal and / or metalloid oxides, wherein the atomic ratio ((i):(ii)) is at least about 1:10 and not more than about 10:1. The mixed oxide-coated particles are preferably synthesized by a vapor-phase coating technique, such as atomic layer deposition. The formulation may provide delayed or sustained release of the glucagon-like peptide-1 receptor agonist for treating metabolic disorders or conditions, such as type 2 diabetes and / or obesity, without a burst effect. The glucagon-like peptide-1 receptor agonist is preferably liraglutide.

Description

[Technical Field]

[0001] The present invention relates to novel formulations for use in the field of drug delivery, for example, and in particular in the treatment of type 2 diabetes and obesity.

[0002] The listing or discussion of an apparently prior-published document in this specification should not necessarily be taken as an acknowledgment that the document is part of the state of the art or general knowledge. [Background technology]

[0003] Diabetes mellitus is a metabolic disorder characterized by chronic hyperglycemia with impaired carbohydrate, fat, and / or protein metabolism resulting from defects in insulin secretion, insulin action, or both. It was one of the earliest diseases in history, described in Egyptian manuscripts around 1500 BC. However, the importance of insulin to the disease was not determined until the 1920s.

[0004] The most common form of diabetes (approximately 90% of all patients) is type 2 diabetes, which has recently become something of a global pandemic and represents a major healthcare burden: the number of patients diagnosed with this condition is predicted to exceed 500 million worldwide by 2035.

[0005] Type 2 diabetes is characterized by impaired insulin production and secretion by pancreatic beta cells and peripheral tissue insulin resistance.

[0006] Type 2 diabetes typically begins at a later age (e.g., middle age or old age), although rates are known to be increasing in younger people. It is not only associated with shorter life expectancy, but also invariably leads to long-term complications, including cardiovascular disease, end-stage renal disease, lower limb amputation, and blindness.

[0007] The etiology of type 2 diabetes is thought to be primarily related to diet, including excessive consumption and / or insufficient energy expenditure, particularly through exercise. In this regard, type 2 diabetes and obesity are interdependent, and weight loss is associated with improved prognosis in overweight type 2 diabetic patients and obese individuals. Indeed, the increasing global burden of type 2 diabetes is thought to be largely attributable to the rise of obesity, which has itself become something of a global pandemic over recent decades.

[0008] Glucagon-like peptide-1 (GLP-1) is a peptide hormone produced after tissue-specific post-translational processing of the proglucagon peptide. It is secreted by enteroendocrine L cells and specific neurons in the brainstem following food consumption. The amino acid sequence of human GLP-1 is described, inter alia, in Schmidt et al., Diabetologia, 28, 704 (1985), as a 37 amino acid residue peptide.

[0009] Along with glucose-dependent insulinotropic peptide (GIP), GLP-1 is an incretin, i.e., it has the ability to enhance insulin secretion and thereby lower blood glucose levels in a glucose-dependent manner. GLP-1-based therapy is associated with weight loss and a lower risk of hypoglycemia, making it particularly important in the treatment of patients with type 2 diabetes.

[0010] Endogenous GLP-1 is rapidly degraded, with a half-life of only about 2 minutes. This has led to substantial research into the development of GLP-1 receptor agonists, among other things, to increase GLP-1 activity. In this regard, GLP-1 and analogs and fragments thereof (hereinafter collectively referred to as "GLP-1 receptor agonists" or "GLP1RAs") are useful as insulin secretagogues and therefore for the treatment of diabetes (including type 1, particularly type 2 diabetes) and / or obesity.

[0011] Approved GLP1RAs include exenatide (Byetta®, Bydureon®, AstraZeneca), liraglutide (Victoza®, Saxenda®, Novo Nordisk), lixisenatide (Lyxumia®, Adlyxin®, Sanofi), albiglutide (Tanzeum®, GlaxoSmithKline), dulaglutide (Trulicity®, Eli Lilly), semaglutide (Ozempic®, Wegovy®, Novo Nordisk) and tirzepatide (Mounjaro®, Eli Lilly).

[0012] The above-listed GLP1RA-containing products are administered by subcutaneous injection into adipose tissue, usually the abdomen or upper thigh. Existing injectable formulations containing GLP1RA are typically injected once or twice daily (classified as short-acting GLP1Ra) or, still relatively frequently, weekly (long-acting GLP1Ra).

[0013] A review by Sikirica et al. in Diabetes Metab. Syndr. Obes., 10, 403 (2017) found that the percentage of patients reporting experiencing problems with GLP1RA injections was higher than average physician estimates. Frequently reported problems include painful injections and / or inconvenient regularity. The most common complaint reported by patients (56%) and physicians (32.6%) was that oral medications were preferred over injections.

[0014] These factors are known to cause patient adherence problems. Despite the general awareness among patients with type 2 diabetes that non-adherence can lead to significant adverse sequelae in the treatment of the condition, in a recent study reported by Weiss et al. in Patient Prefer. Adherence, 27, 2337 (2020), approximately 50% of patients do not strictly adhere to their prescribed medication regimen.

[0015] Semaglutide is formulated as an oral medication in the form of Rybelus® (Novo Nordisk), but the compound has very low oral bioavailability and must be taken daily on an empty stomach with up to 120 mL of water, with the recommendation to eat within 30 to 60 minutes afterwards, all of which may lead to patient compliance issues as well as an increased risk of adverse events.

[0016] In chronic conditions such as type 2 diabetes or obesity, it would be advantageous to provide an extended-release composition in which the active ingredient is released at a desired and predictable rate in vivo over weeks or months after injection to ensure better patient compliance and a more optimal pharmacokinetic profile wherever possible. Increased patient compliance is expected to contribute to improved overall therapeutic efficacy over time (and thus generally healthier patients), as well as healthcare and societal savings.

[0017] In any event, a more convenient, less frequently injected GLP1RA formulation is expected to be an attractive treatment option for all type 2 diabetes patients currently treated with GLP1RAs, even for those for whom adherence to the prescribed dosing regimen is not an issue.

[0018] For any sustained release composition, it is crucial that the release profile exhibits a minimal initial rapid release of the active ingredient, i.e., a high concentration of drug in the plasma immediately after administration. Such a "burst" release can result in undesirably high concentrations of the active ingredient and can be dangerous for drugs that have a narrow therapeutic window or are toxic at high plasma concentrations.

[0019] For injectable suspensions of active ingredients, it is also important that the size of the suspended particles be controlled so that they can be injected through a needle: if large agglomerated particles are present, they will not only block the needle through which the suspension is injected, but will also not form a stable suspension within the infusion solution (i.e., they will tend to sink to the bottom of the infusion solution instead).

[0020] Thus, there is an unmet clinical need for longer-lasting, more effective, and / or improved drug delivery systems involving GLP1RAs in the treatment of conditions such as type 2 diabetes and obesity.

[0021] Atomic layer deposition (ALD) is a technique used to deposit thin films on solid substrates, including a variety of materials, including organic, biological, and polymeric materials, as well as inorganic materials, such as metal oxides. It enables atomic-scale and near-atomic-scale manufacturing (ACSM) of materials, structures, devices, and systems for a wide range of applications (see, for example, Zhang et al. Nanomanuf. Metrol. 2022, https: / / doi.org / 10.1007 / s41871-022-00136-8). Due to its self-limiting properties, ALD can achieve atomic-level thicknesses controlled solely by adjusting the number of growth cycles. Furthermore, multiple layers can be deposited, and the properties of each layer can be customized at the atomic level.

[0022] Due to its atomic-level control, ALD is used, for example, as a key technology for the production of next-generation semiconductors, or in the atomic-level synthesis of advanced catalysts, and in the precise fabrication of nanostructures, nanoclusters, and single atoms (see, e.g., Zhang et al., supra).

[0023] This technique is typically performed at low pressure and high temperature. Film coatings are produced by alternately exposing a solid substrate in an ALD reactor chamber to vaporized reactants in the gas phase. The substrate can be a silicon wafer, granular material, or small particles (e.g., microparticles or nanoparticles).

[0024] The coated substrate is protected from chemical reaction (degradation) and physical change by the solid coating. ALD can also potentially be used to control the release rate of substrate materials in solvents, thereby potentially being used in the formulation of active pharmaceutical ingredients.

[0025] In ALD, a first precursor, which may be metal-containing, is delivered into the ALD reactor chamber (in a so-called "precursor pulse") and forms an adatom or molecular monolayer on the surface of the substrate. Excess first precursor is then purged from the reactor, and a second precursor, such as water, is then pulsed into the reactor. This reacts with the first precursor, thereby forming, for example, a monolayer of metal oxide on the substrate surface. A subsequent purge pulse is followed by a further pulse of the first precursor, thus initiating a new cycle of the same events (a so-called "ALD cycle").

[0026] The thickness of the film coating is controlled, among other things, by the number of ALD cycles performed.

[0027] In typical ALD processes, only atomic or molecular monolayers are produced during any one cycle, resulting in no discernible physical interfaces between these monolayers, which essentially become a continuum at the surface of the substrate.

[0028] International Patent Application WO2014 / 187995 describes a process in which multiple ALD cycles are performed, after which the resulting coated substrate is periodically removed from the reactor and subjected to a redispersion / agitation step to present new surfaces available for precursor adsorption.

[0029] The agitation step was primarily performed to address a problem observed with nanoparticles and microparticles, namely, particle agglomeration during the ALD coating process, resulting in "pinholes" formed by contact points between such particles. The redispersion / agitation step was performed by placing the coated substrate in water and sonicating, which resulted in deagglomeration and disruption of contact points between individual particles of the coated active material.

[0030] The particles were then returned to the reactor, and the powder ALD coating and powder deagglomeration steps were repeated three times for a total of four series of cycles. This process was found to allow for the formation of coated particles that were nearly pinhole-free (see also Hellrup et al., Int. J. Pharm., 529, 116 (2017)).

[0031] The present inventors have now produced novel injectable compositions containing one or more GLP1RAs, in which microparticles containing one or more GLP1RAs are coated with a specific mixed oxide coating layer using ALD, and the coated particles are suspended in a vehicle. This composition produces a favorable pharmacokinetic profile by releasing the active ingredient over an extended period of time, providing therapeutically effective levels of the drug in the systemic circulation, without any significant initial burst effect. Summary of the Invention

[0032] According to a first aspect of the present invention there is provided a pharmaceutical formulation useful for treating a metabolic disorder or condition comprising a plurality of particles suspended in a carrier system, said particles comprising: (a) have an average diameter based on weight, number, or volume of about 10 nm to about 700 μm; (b)(i) zinc oxide (ZnO); (ii) comprising a solid core comprising one or more GLP1RAs or pharmaceutically acceptable salts thereof at least partially coated with a coating of inorganic material comprising a mixture of one or more other metal and / or metalloid oxides; the atomic ratio ((i):(ii)) is at least about 1:10 and not more than about 10:1; This formulation is hereinafter referred to as the "formulation of the present invention." [Brief explanation of the drawings]

[0033] [Figure 1] FIG. 1 shows the results of coating integrity testing of two batches of coated liraglutide microparticles. [Figure 2]FIG. 2 shows the decrease in drug loading of coated and uncoated particles after gamma irradiation. [Figure 3] FIG. 3 shows a semi-logarithmic plot of mean plasma concentrations (ng / mL) versus time (days) following a single subcutaneous administration of liraglutide to rats at various formulations and dose levels. [Figure 4] Figure 4 shows the mean plasma concentrations in the same study over the first 72 hours. [Figure 5] FIG. 5 shows the in vitro release results for two batches of coated liraglutide microparticles. [Figure 6] FIG. 6 shows a semi-logarithmic plot of mean plasma concentrations (ng / mL) versus time (days) following a single subcutaneous dose of liraglutide to minipigs in various formulations. [Figure 7] FIG. 7 shows the in vitro release results for four batches of coated liraglutide microparticles. [Figure 8] FIG. 8 shows a semi-logarithmic plot of mean plasma concentrations (ng / mL) versus time (7 days) following a single subcutaneous dose of liraglutide in various formulations to rats. [Figure 9] FIG. 9 shows a semi-logarithmic plot of mean plasma concentrations (ng / mL) versus time (42 days) following a single subcutaneous dose of liraglutide in various formulations to rats. DETAILED DESCRIPTION OF THE INVENTION

[0034] Preferably, the atomic ratio ((i):(ii)) is between at least about 1:1 and at most about 6:1.

[0035] Coatings comprising a mixture of zinc oxide and one or more other metal and / or semi-metal oxides are hereinafter referred to as "mixed oxide" coatings or coating material(s).

[0036] It will be appreciated by those skilled in the art that the term "solid" includes any form of matter that retains its shape and density when unconfined and / or in which the molecules are generally as tightly packed as the repulsive forces between them allow. A solid core has at least a solid outer surface onto which a layer of coating material can be deposited. The interior of the solid core may also be solid, or alternatively, may be hollow. For example, if the particles are spray-dried before being placed in a reaction vessel, they may be hollow due to the spray-drying technique.

[0037] The solid core of the formulation of the present invention comprises one or more GLP1RAs or pharmaceutically acceptable salts thereof, and in this respect may consist essentially of said one or more GLP1RAs or said salts thereof, or may comprise said one or more GLP1RAs or said salts thereof together with other excipients or other active ingredients.

[0038] By "consisting essentially of" GLP1RA or a pharmaceutically acceptable salt thereof, we include that the solid core comprises essentially only said one or more GLP1RA or salts thereof, i.e., is free of non-biologically active substances such as excipients, carriers (see below) and other active substances. This means that the core may contain less than about 5%, such as less than about 2%, for example less than about 3%, including less than about 1%, of such other excipients and / or active substances.

[0039] Alternatively, the core comprising one or more GLP1RAs or pharmaceutically acceptable salts thereof may comprise such active ingredients mixed with one or more pharmaceutical ingredients, which may include pharmaceutically acceptable excipients such as adjuvants, diluents, or carriers, and / or may include other biologically active ingredients, such as those described below.

[0040] Non-biologically active adjuvants, diluents, and carriers that may be employed in cores coated according to the present invention may include pharmaceutically acceptable substances that are soluble in water, such as carbohydrates, e.g., sugars such as lactose and / or trehalose, and sugar alcohols such as mannitol, sorbitol, and xylitol, or pharmaceutically acceptable inorganic salts such as sodium chloride. Preferred carrier / excipient materials include sugars and sugar alcohols.

[0041] Such excipients are preferably incorporated into the core comprising GLP1RA or a pharmaceutically acceptable salt thereof via the process of spray drying, which will be understood by those skilled in the art to include any method of producing a dry powder from a liquid, including a solution or suspension (including a slurry), which involves using hot gases to rapidly dry and convert the liquid stream into solid particles comprising vaporized solvent and solutes previously dissolved in the solution and / or particles previously suspended in the evaporated liquid.

[0042] GLP1RA and its pharmaceutically acceptable salts can be presented in a crystalline, partially crystalline, and / or amorphous state. Regardless of the physical form, GLP1RA or its pharmaceutically acceptable salts can be in or converted to a solid state at about room temperature (e.g., about 18°C) and about atmospheric pressure. The active agent (and optionally other pharmaceutical ingredients described herein) should also remain in solid form while being coated, for example, in the ALD reactor, and should not physically or chemically degrade to a significant extent (i.e., about 10% w / w or less) while being coated or after being covered by the mixed metal oxide coating material.

[0043] Pharmaceutically acceptable salts of GLP1RA include addition salts, particularly acid addition salts.Such salts can be formed by conventional means, for example, by reacting the free acid or free base form of the active ingredient with one or more equivalents of a suitable base, or preferably an acid, optionally in a solvent or in a medium in which the salt is insoluble, and then removing the solvent or medium using standard techniques (for example, by vacuum, lyophilization or filtration).Salts can also be prepared by techniques known to those skilled in the art, for example, by exchanging the counterion of the active ingredient in the form of a salt with another counterion using a suitable ion exchange resin.

[0044] Particular salts that may be mentioned include, for example, acid addition salts with hydrochloric acid, L-lactic acid, phosphoric acid, (+)-L-tartaric acid, citric acid, propionic acid, butyric acid, hexanoic acid, L-aspartic acid, L-glutamic acid, succinic acid, ethylenediaminetetraacetic acid (EDTA), maleic acid, acids, especially acetic acid, more especially trifluoroacetic acid, and the like.

[0045] Preferred GLP1RAs include exenatide, lixisenatide, albiglutide, dulaglutide, tirzepatide, more preferably semaglutide, especially liraglutide, all of which are known in the prior art and can be synthesised by classical solution phase techniques, solid phase methods or recombinant peptide production, respectively.

[0046] The formulations of the present invention comprise a pharmacologically effective amount of one or more GLP1RAs or pharmaceutically acceptable salts thereof. Preferably, the solid core of the formulations of the present invention comprises a pharmacologically effective amount of one or more (preferably one) GLP1RAs or salts thereof.

[0047] The term "pharmacologically effective amount" refers to that amount of the relevant GLP1RA (alone or in combination) or salt(s) thereof, which is capable of producing a desired physiological change (such as a therapeutic effect) in a treated patient, whether administered alone or in combination with another active ingredient (which may or may not be another GLP1RA). Such biological or medical response or effect in a patient may be subjective (i.e., the subject gives an indication of or feels an effect), may include at least partial alleviation of the symptoms of the disease or disorder being treated, or a cure or prevention of said disease or disorder, and may be objective (i.e., measurable by some test or marker).

[0048] Therefore, the dosage of GLP1RA / salt thereof that may be administered to a patient should be sufficient to affect a therapeutic response over a reasonable and / or relevant time frame. Those skilled in the art will recognize that the selection of the exact dose and composition and the most appropriate delivery regimen will also be influenced not only by the pharmacological properties of the active ingredient and the components of the formulation, but also by, among other factors, the route of administration, the nature and severity of the condition being treated, the physical condition and mental acuity of the recipient, and the age, condition, weight, sex, and response of the patient being treated, the stage / severity of the disease, and genetic differences between patients.

[0049] The dosage of the GLP1RA / salt thereof may also be determined by the timing and frequency of administration. In any event, a medical professional or other skilled artisan can routinely determine the actual dosage of the relevant GLP1RA(s) and / or salt(s) thereof that is / are most suitable for an individual patient.

[0050] The GLP1RA(s) are released over an extended period of time, which can be at least about 3 days, e.g., about 5 days or about 7 days, and up to about 1 year, e.g., up to about 10 months, up to about 8 months, up to about 6 months, up to about 4 months, up to about 3 months, or up to about 4 weeks, e.g., up to about 3 weeks (e.g., up to about 2 weeks).

[0051] Therefore, an appropriate dose of GLP1RA or a pharmaceutically acceptable salt thereof in the formulation of the present invention will provide an exposure (e.g., AUC ) that provides at least the same therapeutic effect as / to those obtained for current, commercially available subcutaneous injections of the relevant GLP1RA(s) used in clinical practice. 最後 (area under the plasma concentration versus time curve to the last detectable concentration over an extended period of time), or more preferably, AUC ∞ A plasma concentration-time profile providing an AUC (defined as the area under the plasma concentration versus time curve to infinity) can be provided.

[0052] The formulations of the present invention are administered by injection (e.g., daily or weekly) of the relevant GLP1RA and are intended to reduce the total exposure (AUC) obtained from the relevant current standard of care / dosing regimen when administered by injection, e.g., intravenously, intramuscularly, or preferably subcutaneously. ∞ ) for any one of the above periods, which is about 80% to about 125% or less of AUC ∞ Exposure may be provided regarding:

[0053] More preferably, the total exposure (e.g., AUC ∞ ) is the total exposure (e.g., AUC ) obtained from the current standard of care / dosing regimen administered by injection of the relevant GLP1RA. ∞ ), at least about 50% (e.g., at least about 65%), at least about 75% (e.g., at least about 80%), such as at least about 85% of the total.

[0054] This allows for a dosage of the relevant GLP1RA or a pharmaceutically acceptable salt thereof within the formulations of the invention to provide, or be capable of providing, a daily or weekly dosage (i.e., the average dosage released per day or week from the formulation after injection over any of the above time periods) that is in the range of about 10% (e.g., about 15%) to about 80% (e.g., about 70%, e.g., about 65%) of the daily or weekly dosage (as needed) administered within the current standard of care, i.e., daily or weekly injection therapy (as needed).

[0055] Thus, the total dose that can be injected into a patient with a formulation of the present invention comprising liraglutide or a pharmaceutically acceptable salt thereof within the formulation of the present invention can range from about 15 mg (e.g., about 270 mg) to a maximum of about 1000 mg.

[0056] Thus, the total dose that may be injected into a patient by a formulation of the invention comprising semaglutide or a pharmaceutically acceptable salt thereof within the formulation of the invention may range from about 1 mg (e.g., about 30 mg) up to about 100 mg.

[0057] The formulations of the present invention provide a steady state release of the relevant GLP1RA after injection, which is expressed as a mean C 最大 This means that the mean C (maximum concentration observed in the plasma concentration versus time curve) is lower than that obtained from the current standard of care / dosing regimen administered by injection of the relevant GLP1RA (e.g., 20±15 ng / mL for liraglutide and 210±30 ng / mL for semaglutide). For formulations of the invention containing liraglutide, the mean C 最大 In the case of a formulation of the present invention containing semaglutide, the mean C 最大 can be about 75 to about 410 ng / mL.

[0058] The GLP1RA-containing core of the formulation of the present invention is provided in the form of nanoparticles, or more preferably microparticles, preferably having an average diameter by weight, number, or volume of about 50 nm (e.g., about 100 nm, e.g., about 250 nm) to about 30 μm, e.g., about 500 nm to about 100 μm, more specifically, about 1 μm (e.g., about 5 μm, including about 7 μm, about 9 μm, about 10 μm, or about 15 μm) up to about 50 μm, e.g., about 25 μm, e.g., about 20 μm.

[0059] As used herein, the term "weight-based average diameter" will be understood by those skilled in the art to include the average particle size characterized and defined from a particle size distribution by weight, i.e., a distribution in which the existing fraction (relative amount) in each size class is defined as the weight fraction obtained, for example, by sieving (e.g., wet sieving). As used herein, the term "number-based average diameter" will be understood by those skilled in the art to include the average particle size characterized and defined from a particle size distribution by number, i.e., a distribution in which the existing fraction (relative amount) in each size class is defined as the number fraction measured, for example, by microscopy. As used herein, the term "volume-based average diameter" will be understood by those skilled in the art to include the average particle size characterized and defined from a particle size distribution by volume, i.e., a distribution in which the existing fraction (relative amount) in each size class is defined as the volume fraction measured, for example, by laser diffraction. Those skilled in the art will also understand that there are other suitable ways of expressing average diameter, such as area-based average diameter, and these other expressions of average diameter are interchangeable with those used herein. Other instruments well known in the art can be employed to measure particle size, such as, for example, instruments sold by Malvern Instruments, Ltd (Worcestershire, UK) and Shimadzu (Kyoto, Japan).

[0060] The particles may be spherical, i.e., they may possess an aspect ratio of less than about 20, more preferably less than about 10, e.g., less than about 4, especially less than about 2, and / or may have a variation in radius (measured from the center of gravity to the particle surface) of at least about 90% of the particles that is not more than about 50% of the average value, e.g., not more than about 30% of that value, e.g., not more than about 20% of that value.

[0061] Nevertheless, the present invention also allows for the coating of particles of any shape. For example, irregularly shaped (e.g., "raisin" shaped), needle-shaped, flake-shaped, or cubic-shaped particles can be coated. For non-spherical particles, the size can be expressed, for example, as the size of a corresponding spherical particle of the same weight, volume, or surface area. Hollow particles, as well as particles with pores, interstices, etc., such as fibrous or "entangled" particles, can also be coated in accordance with the present invention.

[0062] The particles may be obtained in a form suitable for them to be coated, or in that form, for example, by a particle size reduction process (e.g., crushing, cutting, milling, or grinding to a particular weight-based average diameter (defined herein), for example, by utilizing wet grinding, dry grinding, air jet milling (including cryogenic micronization), ball milling such as planetary ball milling, as well as end runner mills, roller mills, vibratory mills, hammer mills, roller mills, fluid energy mills, pin mills, and the like. Alternatively, particles can be directly prepared to a suitable size and shape by, for example, spray drying, freeze drying, spray freeze drying, vacuum drying, precipitation including the use of supercritical fluids, or other top-down methods (i.e., reducing large particle sizes, for example, by grinding), or bottom-up methods (i.e., increasing small particle sizes, for example, by sol-gel techniques, crystallization, and the like). Alternatively, nanoparticles can be made by well-known techniques such as gas condensation, attrition, chemical precipitation, ion implantation, pyrolysis, hydrothermal synthesis, and the like.

[0063] To remove impurities that may result from the production of the particles, the particles may need to be washed and / or purified, and then dried (depending on how the core-containing particles were originally prepared). Drying can be carried out by many techniques known to those skilled in the art, including evaporation, spray drying, vacuum drying, freeze drying, fluidized bed drying, microwave drying, IR radiation, drum drying, etc. Once dried, the cores can then be deagglomerated by grinding, screening, milling, and / or dry sonication. Alternatively, the cores can be treated to remove any volatile materials that may be absorbed onto their surface, for example, by exposing the particles to vacuum and / or elevated temperatures.

[0064] The surface of the core may be chemically activated prior to applying the first layer of coating material, for example, by treatment with hydrogen peroxide, ozone, a free-radical-containing reactant, or by applying a plasma treatment to generate free oxygen radicals on the surface of the core, which may create favorable adsorption / nucleation sites on the core for (e.g., ALD) precursors.

[0065] Thus, the GLP1RA-containing core is at least partially coated with a coating material comprising a mixture of zinc oxide and one or more other metal and / or metalloid oxides, in an atomic ratio of zinc oxide to other oxide(s) of at least about 1:10 (e.g., at least about 1:6 or about 1:4, e.g., at least about 1:2), preferably at least about 1:1 (e.g., at least about 1.5:1, e.g., at least about 2:1) (including at least about 2.25:1, e.g., at least about 2.5:1 (e.g., at least about 3.25:1 or at least about 2.75:1 (including 3:1)), and up to (i.e., less than) about 10:1 (including up to about 8:1, about 7:1, about 6:1, or 5.5:1), or up to about 5:1, e.g., up to about 4.5:1 (including up to about 4:1) (e.g., up to about 3.75:1).

[0066] Preferred methods for applying the coating(s) to the core containing the biologically active agent include gas phase techniques such as ALD or related techniques such as atomic layer epitaxy (ALE), molecular layer deposition (MLD, a technique similar to ALD except that molecules (usually organic molecules) are deposited in each pulse instead of atoms), molecular layer epitaxy (MLE), chemical vapor deposition (CVD), atomic layer CVD, molecular layer CVD, physical vapor deposition (PVD), sputtering PVD, reactive sputtering PVD, evaporation PVD, and binary reaction sequence chemistry. ALD is the preferred coating method according to the present invention.

[0067] When ALD is used, the above-mentioned mixed oxide coatings can be prepared by supplying a first zinc-, other metal-, or metalloid-containing precursor to the ALD reactor chamber (in a so-called "precursor pulse") to form a zinc-, other metal-, or metalloid-containing monolayer of atoms or molecules adsorbed on the surface of the particles. A second precursor (e.g., water) is then pulsed into the reactor and reacts with the first precursor to form a monolayer of zinc, metal, or semimetal oxide, respectively, on the substrate surface. A subsequent purge pulse is followed by another pulse of the first precursor, thus initiating a new cycle of the same events (a so-called "ALD cycle").

[0068] When MLD is used, organic coatings, polymeric coatings and / or hybrid organic-inorganic coatings may be prepared by using MLD precursors, such as organic molecules containing difunctional groups, such as diols, diamines, diisocyanates, dichlorides, dialdehydes, and the like.

[0069] In most cases, the first of the series of reactions involves some functional group or free electron pair or radical on the surface to be coated, such as a hydroxy group (-OH) or a primary or secondary amino group (-NH or -NHR, where R is an aliphatic group, e.g., an alkyl group). Each reaction is advantageously carried out separately under conditions such that essentially all excess reagents and reaction products are removed before carrying out the subsequent reaction.

[0070] To produce a mixed oxide coating in which the atomic ratio of zinc oxide to one or more other metal and / or metalloid oxides (in terms of the number of monolayers applied) is between (for example) about 1:1 and up to about 6:1, one skilled in the art will understand that for every one ALD cycle (i.e., monolayer) of the other oxide(s), about one to about six ALD cycles of zinc oxide must also be deposited. For example, if a mixed oxide coating with a 3:1 atomic ratio (zinc:other oxide) is to be formed, three zinc-containing precursor pulses, each followed by a second precursor pulse, form three monolayers of zinc oxide, then one pulse of the other metal and / or metalloid-containing precursor, followed by a second precursor pulse, form one monolayer of the other metal and / or metalloid oxide. Alternatively, six monolayers of zinc oxide can be followed by two monolayers of the other oxide, or any other combination, to provide an overall atomic ratio of about 3:1. In this regard, the order of pulses to produce the relevant oxides is not important as long as the resulting atomic ratios ultimately fall within the relevant ranges.

[0071] Metal and / or metalloid elements other than zinc that may be mentioned include alkali metals, alkaline earth metals, noble metals, transition metals, post-transition metals, lanthanides, etc. Metals and metalloids that may be mentioned include aluminum, titanium, magnesium, iron, gallium, zirconium, niobium, hafnium, tantalum, lanthanum, and / or silicon, more preferably aluminum, titanium, magnesium, iron, gallium, and / or zirconium. Specific metal and / or metalloid elements that may be mentioned include aluminum and silicon.

[0072] In this regard, the mixed oxide coating material preferably comprises one or the other of aluminum oxide (Al2O3) and / or silicon dioxide (SiO2), or both.

[0073] A method for preparing a plurality of coated particles according to the present invention is provided, wherein the coated particles are made by applying, by vapor deposition techniques, precursors of at least two metal and / or semi-metal oxides that form a mixed oxide onto a solid core and / or a previously coated solid core. Precursors for forming metal or semi-metal oxides often include water, oxygen, ozone, and / or oxygen precursors such as hydrogen peroxide, and metal and / or semi-metal compounds, typically organometallic or organometallic compounds.

[0074] As non-limiting examples of precursors, the precursor of zinc oxide can be water and a di-C1-C5 alkyl zinc such as diethyl zinc. The precursor of aluminum oxide can be water and a tri-C1-C5 alkyl aluminum such as trimethyl aluminum. The precursor of silicon oxide (silica) can be water as the oxygen precursor, as well as silane, alkyl silane, amino silane, and orthosilicate tetraethyl ester. Precursors of iron oxide include oxygen, ozone and water as oxygen precursors, as well as di-C1-C5 alkyl iron, dicyclopropyl iron, and FeCl3. It will be understood that one skilled in the art will recognize which precursors are suitable for the purposes disclosed herein.

[0075] In ALD, the layer of coating material can be applied at a process temperature of about 20°C to about 800°C, or about 40°C to about 200°C, e.g., about 40°C to about 150°C, e.g., about 50°C to about 100°C. The optimal process temperature depends on the reactivity of the precursors and / or materials employed in the core (including biologically active agents, e.g., GLP1RA / salts) and / or the melting point of the core material(s). Lower temperatures, such as about 30°C to about 100°C, are preferably used. In particular, in one embodiment of the present method, a temperature of about 20°C to about 80°C, e.g., about 30°C to about 70°C, e.g., about 40°C to about 60°C, e.g., about 50°C, is employed.

[0076] The inventors have found that when coatings comprising zinc oxide are applied using ALD at low temperatures, such as from about 50°C to about 100°C, the coating material is predominantly crystalline in nature (unlike other coating materials such as aluminum oxide, titanium oxide, and silicon dioxide, which form amorphous layers).

[0077] As alluded to above, without being limited by theory, it is understood that because zinc oxide is crystalline, when zinc oxide alone is employed as a coating material, interfaces may form between adjacent crystals of zinc oxide deposited by ALD, through which a carrier system, vehicle, or solvent (e.g., an aqueous solvent system) in which zinc oxide is partially soluble may penetrate after suspension therein. It is believed that this may result in dissolution that is too rapid for the depot-forming composition that is intended to be made.

[0078] Furthermore, previous studies have shown that formulations containing certain active ingredients coated with zinc oxide have lower relative bioavailability than uncoated active ingredients when suspended in aqueous media. This decrease in relative bioavailability is believed to be due to the degradation of the active ingredient before it can be released into the systemic circulation. It is believed that penetration of water through the crystalline interface within the zinc oxide coating leads to hydrolysis of the active ingredient within the interior of the coated particles.

[0079] It has now been discovered that these problems can be alleviated by creating mixed oxide coatings as described herein. In particular, by forming mixed oxide coatings as described herein that are primarily, if not entirely, composed of zinc oxide, it has been possible to coat active ingredients with a coating that appears to be essentially amorphous, or a composite between crystalline and amorphous materials, and / or that may reduce the penetration of infusion vehicles such as water. In this regard, it appears that the perceived presence of interfaces described above may be reduced or entirely avoided by employing the mixed oxide aspects of the present invention in either a heterogeneous manner (wherein other oxides "fill" the gaps formed by the interfaces) or a homogeneous manner (wherein a true composite of mixed oxide materials is formed during deposition, potentially avoiding interfaces in the first place).

[0080] The vapor deposition reactor chamber used may optionally and / or preferably be a stationary vapor deposition reactor chamber. The term "stationary," in the context of a vapor deposition reactor chamber, will be understood to mean that the reactor chamber remains stationary during use to carry out a vapor deposition technique, excluding negligible movement and / or vibration, such as caused by associated machinery.

[0081] Furthermore, for example, as described below, a so-called "stop-flow" process can be employed. Using the stop-flow process, a first precursor is supplied to a reactor chamber and allowed to contact the core within the reactor chamber for a predetermined period (which can be considered a soak time) before the first precursor is purged from the reactor chamber. During the predetermined period, there is preferably a substantial absence of pumps that can cause gas flow and / or a substantial absence of mechanical agitation of the core.

[0082] The use of a stop-flow process can improve coating uniformity by allowing each gas to conform and diffuse into high-aspect-ratio substrates, such as powders. The advantage can be even more pronounced when using slower-reactive precursors, since the precursors are given more time to react on the surface. This can be particularly evident when depositing mixed oxide coatings according to the present invention. For example, when depositing mixed zinc oxide / aluminum oxide coatings as described herein, we found that zinc-containing precursors, such as diethylzinc (DEZ), have a lower reaction probability toward the substrate surface than aluminum-containing precursors, such as trimethylaluminum (TMA).

[0083] In addition to producing coatings with good shell integrity and more controlled release profiles, employing such a stopped-flow process may improve the ability to achieve specific coating compositions.

[0084] For example, as noted above, when attempting to employ gas phase techniques to produce a coating with a 3:1 atomic ratio between zinc and aluminum in the resulting shell, it was found that a ratio much closer to 3:1 could be achieved using a stopped-flow process than when depositing the material using a continuous flow of precursors.

[0085] Preferably and / or optionally, a "multi-pulse" technique may be employed to deliver the first precursor, the second precursor, or both precursors to the reactor chamber.

[0086] Using such multi-pulse techniques, each precursor can be delivered to the reactor chamber as multiple "sub-pulses," each lasting a short period of time, such as from 1 second to about 1 minute (depending on the size and nature of the vapor deposition reactor), rather than as a single continuous pulse. The precursor may be allowed to contact the core within the reactor chamber for a predetermined period of time, such as from about 1 to 500 seconds, about 2 to 250 seconds, about 3 to 100 seconds, about 4 to 50 seconds, or about 5 to 10 seconds, e.g., 9 seconds, after each sub-pulse. Again, depending on the size and nature of the vapor deposition reactor, this period can extend up to several minutes (e.g., up to about 30 minutes). The introduction of sub-pulses, followed by a period of soak time, can be repeated a predetermined number of times, such as from about 5 to 1000 times, about 10 to 250 times, or about 20 to 50 times, in a single step.

[0087] Formulations of the present invention that may be mentioned include those not prepared by such a stopped flow process.

[0088] The core may be coated with one or more separate, individual layers of a mixed oxide coating as defined herein. Preferably, two or more separate, individual mixed oxide layers, coatings or shells (these terms are used interchangeably herein) are applied sequentially (i.e., "separately applied") to the solid core containing GLP1RA(s).

[0089] The "separate application" of "separate layer, coating, or shell" means that a solid core may be coated with a first layer of coating material, which layer is formed by two or more (e.g., multiple or set of) cycles as described herein, each cycle producing zinc oxide, or other metal and / or semi-metal oxide (as appropriate), and the resulting coated core may then be subjected to some form of deagglomeration process.

[0090] In other words, a "vapor deposition (e.g., ALD) cycle" may be repeated several times to provide a "vapor deposition (e.g., ALD) set" of cycles, which may consist of, for example, 10, 25, or 100 cycles. However, after this series of cycles, the coated cores may be subjected to some form of deagglomeration process, which may be followed by a further series of cycles.

[0091] This process can be repeated as many times as necessary, and thus the number of distinct layers of coating material produced by multiple successive cycles in the final coating corresponds to the number of these intermittent deagglomeration steps, with a final mechanical deagglomeration option occurring prior to application of the final layer (series of cycles) of coating material.

[0092] The particles of the formulation may have from 1 to about 100 individual layers of the mixture of oxides (and, if appropriate, other coating materials as described below), such as from 2 to about 50 individual layers, such as from 3 to about 10 individual layers, such as from 3 to 6 individual layers.

[0093] The terms "deagglomeration" and "deagglomeration" are used interchangeably when referring to coated particles, and deagglomeration of coated particle agglomerates is preferably accomplished by mechanical sieving techniques.

[0094] The coated cores may be subjected to the aforementioned deagglomeration process internally without being removed from the apparatus in a continuous process. Such a process may include forcing the solid product mass formed by coating the cores through a sieve disposed within the reactor, configured to deagglomerate particle agglomerates upon forcing the coated cores by a forcing means applied within the reactor prior to being subjected to a second and / or further coating. This process may be continued as many times as necessary and / or appropriate before applying a final coating, as described herein.

[0095] Having a sieve located within the reactor means that the coating can be applied in a continuous process without the need to remove the particles from the reactor. Therefore, there is no need for manual handling of the particles, and no external machinery is required to deagglomerate agglomerated particles. This not only significantly reduces the time the coating process takes to run, but also makes it more convenient and reduces the risk of personnel handling harmful (e.g., toxic) materials. It also increases the reproducibility of the process by limiting manual handling and reduces the risk of contamination.

[0096] Alternatively and / or preferably, the coated cores can be removed from the coating apparatus, such as an ALD reactor, and then subjected to an external deagglomeration step, as described, for example, in International Patent Application Publication No. 2014 / 187995. Such an external deagglomeration step can include agitation, such as wet or dry sonication, or, preferably, sieving the resulting solid product mass from the reactor, e.g., by passing it through a sieve or mesh, to deagglomerate the particles, e.g., as described below, before returning the particles to the coating apparatus for the next coating step. Again, this process can be continued as many times as necessary and / or appropriate before applying the final coating.

[0097] In the external deagglomeration process, deagglomeration can alternatively (and / or instead of the processes described above) be carried out by subjecting the wet or dry coated particles to one or more of nozzle aerosol generation, milling, grinding, stirring, high shear mixing and / or homogenization. If the deagglomeration step(s) are carried out on wet particles, the deagglomerated particles should be dried (as described above for the cores) before the next coating step.

[0098] However, in such external processes, the deagglomeration step(s) include one or more sieving steps, which may include jet sieving, manual sieving, vibratory sieve shaking, horizontal sieve shaking, tap sieving, or (preferably) sonic sieving as described below, or a similar process involving any combination of these sieving steps. Manufacturers of suitable sonic sifters include Advantech Manufacturing, Endecott, and Tsutsui.

[0099] Vibratory screening techniques may involve vibrating the solid product mass formed by coating the cores through a screen located either internally or (preferably) externally (i.e., outside) the reactor, configured to deagglomerate any particle agglomerates during the vibratory passage of the coated cores prior to subjecting them to a second and / or further layer of coating material, the process being repeated as many times as necessary and / or appropriate before applying a final layer of coating material.

[0100] The vibrational forcing means includes a vibration motor coupled to the sieve. The vibration motor is configured to vibrate and / or rotate when power is supplied to it. For example, the vibration motor can be a piezoelectric vibration motor including a piezoelectric material that changes shape when an electric field is applied as a result of the inverse piezoelectric effect. The change in shape of the piezoelectric material causes acoustic or ultrasonic vibration of the piezoelectric vibration motor.

[0101] Alternatively, the vibration motor may be an eccentric rotating mass (ERM) vibration motor, which includes a mass that rotates when power is applied to the motor. The mass is eccentric from the axis of rotation, and the rotation of the mass causes the motor to become unbalanced, vibrating and / or rotating. Additionally, an ERM vibration motor may include multiple masses positioned at different locations relative to the motor. For example, an ERM vibration motor may include an upper mass and a lower mass, each positioned at opposite ends of the motor. By varying each mass and its angle relative to the other mass, the vibration and / or rotation of the ERM vibration motor can be varied.

[0102] The vibration motor is coupled to the sieve in a manner such that when power is supplied, the vibration and / or rotation of the motor is transmitted to the sieve.

[0103] The sieve and vibrating motor may be suspended from a mount (e.g., a floor-mountable frame) via a suspension means such that the sieve and motor vibrate freely relative to the mount, without vibrations being substantially transmitted to or damped by the mount. This allows the vibrating motor and sieve to vibrate and / or rotate without obstruction and also reduces noise generated during the vibratory sieving process. The suspension means may include one or more springs or bellows (i.e., air cushions or equivalent cushioning means) coupling the sieve and / or motor to the mount. Manufacturers of vibrating sieves or sifters suitable for carrying out such processes include, for example, Russell Finex, SWECO, Filtra Vibration, VibraScreener, Gough Engineering, and Farley Greene.

[0104] Preferably, the vibrating screening technique further comprises controlling a vibrating probe coupled to the sieve, the vibrating probe being controllable to vibrate the sieve at a frequency separate from the frequency of the vibrations caused by the vibration motor, preferably the vibrating probe vibrates the sieve at a higher frequency than the vibrations caused by the vibration motor, more preferably the frequency is in the ultrasonic range.

[0105] Providing additional vibration to the sieve by the vibrating probe reduces the occurrence of sieve clogging, reduces the likelihood of the sieve being overloaded, and reduces the time required to clean the sieve mesh.

[0106] Preferably, the vibratory screening technique involves screening the coated particles at a throughput of at least 1 g / min. More preferably, the vibratory screening technique involves screening the coated particles at a throughput of 4 g / min or greater.

[0107] Throughput depends on the sieve mesh area, sieve mesh size, particle size, particle stickiness, and particle static properties. By combining several of these features, much higher throughputs are possible. Thus, vibratory sieving techniques may more preferably involve sieving coated particles at throughputs of up to 1 kg / min or more.

[0108] Any one of the above throughputs represents a significant improvement over the use of known mechanical sieving or sieving techniques. For example, it has been found that sonic sieving involves sieving in 15-minute periods, with a 15-minute cooling time required to store the equipment. To sieve 20 g of coated particles, nine sets of 15 minutes of active sieving time were required, or a total time (including cooling) of 255 minutes. In comparison, by using the aforementioned vibration sieving technique, 20 g of coated particles can be continuously sieved in a maximum of 20 minutes, or more preferably in only 5 minutes or less.

[0109] The mesh size of the sieve can be determined so that the ratio of the size of the sieved or sonicated particles to the mesh size of the sieve is about 1:>1, preferably about 1:2, and optionally about 1:4. The mesh size of the size can range from about 20 μm to about 100 μm, preferably from about 20 μm to about 60 μm.

[0110] Suitable sieve meshes may include perforated plates, microplates, grids, diamonds, threads, polymers, or wires (woven wire sieves), but are preferably formed from metals such as stainless steel.

[0111] Surprisingly, the use of stainless steel mesh within a vibratory sieving technique is as gentle on particle coatings as the use of softer polymer sieves as part of a mechanical sieving technique such as sonic sieving.

[0112] Also, a known problem with sieving powders is the generation of potentially dangerous static electricity. Steel mesh has the advantage of removing static electricity from the powder, but this is not the case with the polymer mesh that must be used in sonic sifters.

[0113] Furthermore, the mesh size of known sonic sifters is limited to approximately 100 μm because the sound waves move through the mesh rather than vibrating it. That limitation does not exist for use with vibratory sifting techniques, which do not rely on sound waves to generate vibrations in the sieve. Thus, the vibratory sifting techniques described herein allow for the sifting of larger particles than would be possible with alternative mechanical sifting techniques.

[0114] When the sieve is located external to the reactor (i.e., outside the reactor), the process for making the coated cores of the formulations of the present invention includes discharging the coated particles from the vapor deposition reactor before subjecting the coated particles to agitation, and reintroducing the deagglomerated coated particles into the vapor deposition reactor before applying at least one additional layer of coating material to the reintroduced particles.

[0115] As mentioned above, the inventors have found that applying a separate layer of coating material after external deagglomeration results in a visible and discernible interface that can be observed by analyzing coated particles according to the invention, e.g., by TEM, as a region of high electron transparency. In this regard, the thickness of the layer between interfaces directly corresponds to the number of cycles in each series carried out within the ALD reactor and between individual external agitation steps.

[0116] In ALD coating processes, such a clear physical interface is typically more difficult to observe because the coating occurs at the atomic level.

[0117] Without being limited by theory, it is believed that removing the coated particles from the vacuum conditions of the ALD reactor and exposing the newly coated surface to the atmosphere leads to structural rearrangement due to relaxation and restructuring of the outermost atomic layers. Such a process is believed to involve a reorganization of atoms at (and near) the surface, driven by a thermodynamic tendency to reduce the surface free energy.

[0118] Additionally, surface adsorption of species, such as hydrocarbons always present in air, can contribute to this phenomenon, as can surface modifications resulting from reactions of the formed coating with hydrocarbons, as well as atmospheric oxygen, etc. Thus, chemical analysis of such interfaces may contain traces of contaminants not originating from the coating process, such as ALD, or core materials, such as active components that form part of the core.

[0119] Whether performed inside or outside the reactor, particle agglomerates are preferably broken down by forcing them through a sieve, thus separating the agglomerates into individual particles or agglomerates of a desired and predetermined size (thereby achieving deagglomeration). Regarding the latter, in some cases, the individual primary particle sizes are so small (i.e., <1 μm) that it is not possible to achieve "complete" deagglomeration (i.e., agglomerates broken down into individual particles). Instead, deagglomeration is achieved by breaking down larger agglomerates into smaller agglomerates of secondary particles of the desired size, as determined by the mesh size of the sieve. The smaller agglomerates are then coated by gas-phase techniques to form fully coated "particles" in the form of small agglomerate particles. Thus, the term "particles," when referring to deagglomerated and coated particles in the context of the present invention, refers to both individual (primary) particles and agglomerated (secondary) particles of the desired size.

[0120] In either case, the desired particle size (whether it be individual particles or agglomerates of the desired size) is maintained, and furthermore, continued application of the gas phase coating mechanism to the particles after such deagglomeration by sieving will form a complete coating on the particles, thus meaning that fully coated particles (individual or agglomerates of the desired size) are formed.

[0121] The repeated coating and deagglomeration process described above, whether carried out inside or outside the reactor, may be carried out at least once, preferably twice, more preferably three times, such as four times, including five times, more particularly six times, such as seven times, and up to about 100 times, such as up to about 50 times, such as up to about 40 times, including up to about 30 times, for example, 2 to 20 times, for example, 3 to 15 times, for example, 10 times, for example, 9 or 8 times, more preferably 6 or 7 times, especially 4 or 5 times.

[0122] Whether performed inside or outside the reactor, at least one screening step is performed, and preferably includes a vibratory screening step as described above. It is even more preferred that the final screening step includes a vibratory screening step performed prior to the application of the final layer (set of cycles) of coating material. However, it is even more preferred that more than one (including each) of the screening steps include the vibratory screening techniques, steps, or processes described herein.

[0123] The preferred repetition of these steps further benefits the improved throughput of any vibratory screening technique.

[0124] Formulations of the present invention that may be mentioned include those that are not prepared by processes involving vibrating sieve technology.

[0125] The total thickness of the coating (meaning all separate layers / coatings / shells) ranges on average from about 0.5 nm to about 2 μm.

[0126] The minimum thickness of each individual layer / coating / shell is in the range of about 0.1 nm on average (eg, about 0.5 nm, such as about 0.75 nm, such as about 1 nm).

[0127] The maximum thickness of each individual layer / coating / shell will depend on the size of the core (initially) and subsequently on the size of the core with any previously applied coatings, and may be on average about 1 / 100th the mean diameter (i.e., mean diameter based on weight, number, or volume) of the core, or cores to which the previously applied coatings have been applied.

[0128] Preferably, for particles having an average diameter of about 100 nm to about 1 μm, the total coating thickness should average about 1 nm to about 5 nm; for particles having an average diameter of about 1 μm to about 20 μm, the coating thickness should average about 1 nm to about 10 nm; and for particles having an average diameter of about 20 μm to about 700 μm, the coating thickness should average about 1 nm to about 100 nm.

[0129] In this regard, the coated cores of the formulations of the present invention preferably have an average diameter, by weight, number or volume, of about 50 nm (e.g., about 100 nm, e.g., about 250 nm) to about 30 μm, e.g., about 500 nm to about 100 μm, more particularly about 1 μm (e.g., about 5 μm, including about 7 μm, about 9 μm, about 10 μm, or about 15 μm) up to about 50 μm, e.g., about 25 μm, e.g., about 20 μm.

[0130] It has been found that applying a coating / shell followed by one or more deagglomeration steps, such as sonication, can result in wear, pinholes, breaks, gaps, cracks, and / or voids (hereinafter "cracks") in the layer / coating because the coated particles are essentially more tightly "bonded" or "glued" together immediately after applying a thicker coating, which can expose the core containing the biologically active component to elements once deagglomeration occurs.

[0131] For example, if it is intended that the particles be provided in a suspension prior to administration to a patient, it is necessary to provide deagglomerated primary particles without pinholes or cracks in the coating, which would result in an undesirable initial peak (burst) in the plasma concentration of the active ingredient immediately after administration.

[0132] We have discovered that performing one or more of the deagglomeration steps described herein results in significantly fewer pinholes, gaps, or cracks in the final layer of coating material, resulting in particles that are not only completely coated with that layer / coating, but are also coated in a manner that allows the particles to be easily deagglomerated (e.g., using non-invasive techniques such as vortexing) in a manner that does not disrupt the layer of coating material formed prior to and / or during pharmaceutical formulation.

[0133] In this regard, the mixed oxide coating typically completely surrounds, encloses, and / or encapsulates the solid core containing the active ingredient(s). In this way, the risk of an initial drug concentration burst due to direct drug contact with a solvent in which the associated active ingredient is soluble is minimized. This may include not only bodily fluids, but also any medium in which such coated particles may be suspended prior to injection.

[0134] Thus, in a further embodiment of the present invention, there is provided a particle as disclosed herein, wherein the coating surrounding, surrounding and / or encapsulating the core covers at least about 50%, such as at least about 65%, for example, at least about 75%, such as at least about 80%, more particularly at least about 90%, such as at least about 91%, for example, at least about 92%, such as at least about 93%, for example, at least about 94%, such as at least about 95%, for example, at least about 96%, such as at least about 97%, for example, at least about 98%, such as at least about 99%, such as approximately or about 100% of the surface of the core, wherein the coating essentially completely surrounds, surrounds and / or encapsulates the core.

[0135] As used herein, the term "essentially completely coating completely surrounding, enclosing, and / or encapsulating the core" means coverage of at least about 98%, or at least about 99%, of the surface of the solid core.

[0136] Alternatively, the processes described herein may result in deagglomerated coated particles that are essentially free of such cracks through which the active ingredient may be released in an uncontrolled manner.

[0137] Although some minor cracks may appear in the coating without affecting its essential function in terms of controlling release, in a further embodiment, there is provided a particle as previously disclosed herein wherein at least about 90% of the particles exhibit no cracks in the coating surrounding, enclosing, and / or encapsulating the core. In one embodiment, at least about 91%, such as at least about 92%, for example, at least about 93%, such as at least about 94%, for example, at least about 95%, such as at least about 96%, for example, at least about 97%, for example, at least about 98%, for example, at least about 99%, such as approximately 100% of the particles exhibit no cracks.

[0138] Alternatively, "essentially free of cracks" in the coating(s) means that less than about 1% of the surface of the coated particle contains wear, pinholes, breaks, gaps, cracks and / or voids (through which the active ingredient is potentially exposed (e.g., to the elements)).

[0139] The layer of coating material can be collectively of essentially uniform thickness across the surface area of ​​the particle. By "essentially uniform" thickness is meant that the degree of variation in coating thickness of at least about 10%, e.g., about 50%, such as about 25% of the coated particles present in the formulations of the invention is about ±20% or less (including ±50% or less) of the average thickness as measured by TEM.

[0140] In addition to the essential mixed oxide coating employed in the formulations of the present invention, other coating materials may also be applied either between separate mixed oxide coatings (e.g., during a separate deagglomeration step) and / or while the mixed oxide coatings herein are being applied, which may be pharmaceutically acceptable and essentially non-toxic coating materials. Such materials may include multiple layers or composites of the mixed oxide and one or more different inorganic or organic materials to modify the properties of the layer(s).

[0141] The additional coating material may include organic or polymeric materials such as polyamides, polyimides, polyureas, polyurethanes, polythioureas, polyesters, or polyimines. The additional coating material may also include hybrid materials (such as those between organic and inorganic materials), including materials that are combinations of metals or other elements with alcohols, carboxylic acids, amines, or nitriles. Such additional organic coatings, polymeric coatings, and / or hybrid organic-inorganic coatings are preferably applied using coating techniques including MLD, as described above. Such polymeric coatings may be polyimides, polyazomethines, polyureas, polyamides, nylons, metal cones, alucones, titanium cones, zincon, metal-organic framework polymers, oxycarbides, and hybrid nanolaminates.

[0142] However, it is preferred that the coating material comprises an inorganic material.

[0143] Additional inorganic coating materials (to zinc oxide) may include other compounds of metals and / or metalloids such as oxides, nitrides, sulfides, selenides, carbonates, other ternary compounds, etc. Metals and metalloids, hydroxides, especially oxides, especially metal oxides are preferred.

[0144] Furthermore, oxides of elements other than zinc, aluminum or silicon that may be mentioned include alkali metals, alkaline earth metals, noble metals, transition metals, post-transition metals, lanthanides, etc. Metals and metalloids that may be mentioned include titanium, magnesium, iron, gallium, zirconium, niobium, hafnium, tantalum and / or lanthanum, more preferably titanium, magnesium, iron, gallium and / or zirconium.

[0145] Thus, additional coating materials that may be mentioned include titanium dioxide (TiO2), iron oxide (Fe x O y, for example, FeO and / or Fe2O3 and / or Fe3O4), gallium oxide (Ga2O3), magnesium oxide (MgO), niobium oxide (Nb2O5), hafnium oxide (HfO2), tantalum oxide (Ta2O5), lanthanum oxide (La2O3), zirconium dioxide (ZrO2), and / or silicon dioxide (SiO2). More preferred additional coating materials include iron oxide, titanium dioxide, zinc sulfide, and more preferably one or both of aluminum oxide (Al2O3) and / or silicon dioxide (SiO2).

[0146] We prefer that the additional inorganic material comprises one or more layers of a single metal oxide (e.g., titanium oxide, silicon oxide, or preferably, aluminum oxide), applied to the core before and / or after application of the mixed oxide layer as described herein.

[0147] Although the particles coated with the multiple mixed oxides according to the present invention are essentially free of the aforementioned cracks in the applied coating through which the active ingredient is potentially exposed (e.g. to the elements), two further optional steps may be applied to the multiple coated particles before subjecting them to further processing in a pharmaceutical formulation.

[0148] The first optional step may involve the application of a final overcoating layer, after the final deagglomeration step previously described herein, where the thickness of that outer "overcoating" layer / coating, or "sealing shell" (these terms are used interchangeably herein), must be thinner than any previously applied separate layer / coating / shell (or "subshell").

[0149] Thus, the thickness can be about 0.7 times (e.g., about 0.6 times) or less, on average, the thickness of the widest previously applied subshell. Alternatively, the thickness can be about 0.7 times (e.g., about 0.6 times) or less, on average, the thickness of the last subshell applied and / or about 0.7 times (e.g., about 0.6 times) or less, on average, the average thickness of all previously applied subshells. The thickness can be in the range of about 0.3 nm to about 10 nm, on average, for particles up to about 20 μm. For larger particles, the thickness can be about 1 / 1000 or less, on average, of the average diameter based on weight, number, or volume of the coated particles.

[0150] The role of the sealing shell is to provide a "sealing" overcoating layer on the particles, covering their cracks, resulting in particles that are not only completely coated with their sealing shell, but also coated in a manner that allows the particles to be easily deagglomerated (e.g., using non-invasive techniques such as vortexing) in a manner that does not disrupt the underlying subshells formed before and / or during pharmaceutical formulation.

[0151] For reasons described herein, the sealing shell preferably does not contain zinc oxide, although it may contain silicon dioxide or, more preferably, aluminum oxide.

[0152] This second optional step may involve subjecting the few remaining particles with broken and / or cracked shells / coatings to a treatment in which all particles are suspended in a solvent in which the active ingredient is soluble (e.g., with a solubility of at least about 0.1 mg / mL) while the least soluble material of the mixed oxide coating is insoluble (e.g., with a solubility of about 0.1 μg / mL or less), followed by separation of the solid particles from the solvent, for example by centrifugation, sedimentation, flocculation, and / or filtration, thereby ensuring that primarily intact particles remain.

[0153] The above optional step provides a means of further potentially reducing the likelihood of a (possibly) undesirable initial peak (burst) in plasma concentration of the active ingredient, as previously discussed herein.

[0154] At the end of the process, the coated particles may be dried using one or more of the techniques previously described herein for drying cores. Drying may occur in the absence or presence of one or more pharmaceutically acceptable excipients (e.g., sugars or sugar alcohols).

[0155] Alternatively, at the end of the process, the separated particles may be resuspended in a solvent (e.g., water, with or without the presence of one or more pharmaceutically acceptable excipients as defined herein) for subsequent storage and / or administration to a patient.

[0156] Prior to applying the first layer of coating material, or between successive coatings, the core and / or partially coated particles may be subjected to one or more alternative and / or preliminary surface treatments. In this regard, one or more intermediate layers comprising a different material (i.e., other than inorganic material(s)) may be applied to the relevant surface, for example, to protect the core or partially coated particles from undesired reactions with precursors during the coating step(s) / deposition process, to enhance coating efficiency, or to reduce agglomeration.

[0157] The intermediate layer may include one or more surfactants, for example, to reduce agglomeration of the coated particles and to provide a hydrophilic surface suitable for subsequent coating. Suitable surfactants in this regard include well-known nonionic, anionic, cationic, or zwitterionic surfactants, such as the Tween series, e.g., Tween 80. Alternatively, the core may undergo a preliminary surface treatment if the active ingredient employed as part of (or as) the core is susceptible to reaction with one or more precursor compounds that may be present in the gas phase during the coating (e.g., ALD) process.

[0158] Application of an "intermediate" layer / surface treatment of this nature may alternatively be achieved by liquid phase non-coating techniques followed by freeze drying, spray drying, or other drying methods to provide particles with a surface layer onto which a coating material can later be applied.

[0159] The outer surface of the particles of the formulations of the invention can also be derivatized or functionalized with chemical compounds or moieties that enhance targeted delivery of the particles within a patient to which the nanoparticles are administered, e.g., by attaching one or more chemical compounds or moieties to the outer surface of the final layer of coating material. Such compounds can be organic molecules (e.g., PEG), polymers, antibodies or antibody fragments, or receptor-binding proteins or peptides, etc.

[0160] Alternatively, the moiety can be an anchoring group, such as a moiety containing silane functionality (see, e.g., Herrera et al., J. Mater. Chem., 18, 3650 (2008) and U.S. Pat. No. 8,097,742). Another compound, e.g., a desired targeting compound, can be attached to such an anchoring group by a covalent bond or a non-covalent bond, including a hydrogen bond or a van der Waals bond, or a combination thereof.

[0161] The presence of such anchoring groups can provide a versatile tool for targeted delivery to specific sites in the body. Alternatively, the use of compounds such as PEG can allow particles to circulate in the bloodstream for longer periods of time, ensuring that they do not accumulate in the liver or spleen (the body's natural mechanisms for eliminating particles, which can prevent delivery to diseased tissues).

[0162] The core coated with the mixed oxide coating, whether in the form of a separate, discrete layer, coating, or shell, as defined herein, is hereinafter referred to as the "coated particle of the formulation of the present invention."

[0163] According to a further aspect of the invention, the coated particles of the formulation of the invention comprise a plurality of particles, the particles comprising: (a) have an average diameter based on weight, number, or volume of about 10 nm to about 700 μm; (b)(i) zinc oxide (ZnO); (ii) comprising a solid core comprising one or more GLP1RAs or pharmaceutically acceptable salts thereof at least partially coated with a coating of inorganic material comprising a mixture of one or more other metal and / or metalloid oxides; The atomic ratio ((i):(ii)) is at least about 1:10 and not more than about 10:1.

[0164] For the avoidance of doubt, all aspects, including preferred aspects, of the formulations of the present invention disclosed and / or claimed herein are equally applicable as aspects and / or preferences of the coated particles of the formulations of the present invention. For the further avoidance of doubt, all such aspects, preferences and features, singly or in combination, are hereby incorporated by reference into this aspect of the invention.

[0165] Pharmaceutical (or veterinary) formulations of the compositions of the invention may contain different types of particles, for example, particles with different functionalization (as described above), particles of different sizes and / or different thicknesses of layers of mixed oxide coating materials, or combinations thereof. By combining particles with different coating thicknesses and / or different core sizes in a single pharmaceutical formulation, drug release after administration to a patient can be controlled (e.g., varied or extended) over a specific period of time.

[0166] The formulations of the present invention may be administered systemically, for example, by infusion, intravenously or intra-arterially (including by intravascular or other perivascular devices / formulations (e.g., stents)), intraosseously, intracerebrally, intraventricularly, intrasynovially, intrasternally, intrathecally, intralesionally, intracranially, intratumorally, cutaneously, intradermally, transdermally, or most preferably, by injection in the form of a pharmaceutically (or veterinarily) acceptable dosage form, for example, intramuscularly or preferably subcutaneously.

[0167] Preparation of the formulations of the present invention involves incorporating the coated particles described herein into a suitable pharmaceutically acceptable carrier system (we include physiologically acceptable carrier systems), which can be accomplished taking into account the intended route of administration and standard pharmaceutical practice. Therefore, suitable excipients (including, for example, physiologically acceptable injectable, e.g., physiologically acceptable intramuscularly injectable, or more preferably, physiologically acceptable subcutaneously injectable excipients, as described herein) should be chemically inert to the active agent used and have no adverse side effects or toxicity under the conditions of use. Such pharmaceutically acceptable carriers may also impart immediate or modified release of the active agent from the particles of the formulations of the present invention.

[0168] Thus, the pharmaceutically acceptable carrier system according to the present invention may be oil-based or oil-based. Thus, the carrier system may comprise one or more pharmaceutically or veterinarily acceptable liquid lipids, which may include fixed oils such as monoglycerides, diglycerides, or triglycerides, including Miglyol™ (e.g., 812N), propylene glycol dicaprylocaprate (Miglyol 840, C8 / C10 ester), tricaprylin (Miglyol oil), Gelucire™ 43 / 01, Kollisolv™ GTA, and Labrafil™. Carrier systems may also include polysorbates such as polysorbate 20, polysorbate 60, and polysorbate 80; glycols such as propylene glycol, polyethylene glycol, polyethylene glycol 300, polyethylene glycol 400, and polyethylene glycol 600; and / or natural and / or refined pharmaceutically acceptable oils such as olive oil, peanut oil, soybean oil, corn oil, cottonseed oil, sesame oil, castor oil, oleic acid, and polyoxyethylated versions thereof (e.g., sorbitan trioleate, Lauroglycol 90, Capryol™ PGMC, PEG-60 hydrogenated castor oil, polyoxyl 35 castor oil). Non-lipid oils such as ethyl lactate and perfluorohexyl octane (F6H8) can also be used. More preferred carrier systems include mono-, di-, and / or triglycerides, including alkyl chain triglycerides (e.g., C6-C 12 Medium chain triglycerides, such as alkyl chain triglycerides, are most preferred.

[0169] Sterile aqueous suspensions of particles of the formulation of the present invention can be formulated according to techniques known in the art. The aqueous medium should contain at least about 50% water but may include other aqueous excipients such as Ringer's solution, and may also include polar cosolvents (e.g., ethanol, glycerol, propylene glycol, 1,3-butanediol, polyethylene glycols of various molecular weights, and tetraglycols), viscosity-increasing or thickening agents (e.g., carboxymethylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, ethylhydroxyethylcellulose, sodium starch glycolate, poloxamers such as poloxamer 407, polyvinylpyrrolidone, cyclodextrins such as hydroxypropyl-β-cyclodextrin, polyvinylpyrrolidone, and polyethylene glycols of various molecular weights), surfactants / wetting agents to achieve a homogeneous suspension (e.g., sorbitan esters, sodium lauryl sulfate, monoglycerides, polyoxyethylene esters, polyoxyethylene alkyl ethers, polyoxylglycerides, and preferably Tweens (polysorbates) such as Tween 80 and Tween 20). Preferred ingredients include tonicity adjusting agents (e.g., sodium lactate, dextrose, especially sodium chloride), pH adjusting agents and / or buffering agents (e.g., citric acid, sodium citrate, especially phosphate buffers such as disodium hydrogen phosphate hydrate, sodium phosphate, sodium dihydrogen phosphate monohydrate, and combinations thereof, which may be employed in combination with standard inorganic acids and bases such as hydrochloric acid and sodium hydroxide), and other ingredients such as mannitol, croscarmellose sodium, and hyaluronic acid.

[0170] The formulations of the present invention can further be formulated in the form of an injectable suspension of coated particles having a size distribution that can form (or can be reconstituted after an appropriate degree of agitation) a uniform and stable (i.e., non-settling) suspension in an injection solution so that they may be injected through a needle. In this regard, the formulations of the present invention can contain inactive ingredients that may prevent premature "solidification" (i.e., the formation of a solid or semi-solid, non-dispersible residue) or gelation (e.g., hydrogel formation) within the formulation, by which we mean that the formulation is sufficiently viscous to prevent settling, so that the suspension is not "uniform" and therefore poses the risk of under- or over-dosing of the active ingredient, or that the formulation can be redispersed to form a sufficiently uniform dispersion before administration.

[0171] Considering the above requirements, the formulation of the present invention, which comprises an aqueous carrier system in which coated particles are suspended, comprises: i. may further comprise a physiologically acceptable injectable (e.g., intramuscularly and / or more preferably subcutaneously injectable) compound capable of reacting with zinc and thereby reducing (e.g., essentially preventing) the reaction of zinc with water to form zinc hydroxide. Such a compound may, for example, comprise a compound that, when dissolved in an aqueous carrier, provides a source of counterion capable of forming a compound (e.g., a salt) with zinc, whose counterion is essentially insoluble in said aqueous medium (at any given temperature, pressure, and pH); ii.1. Inherently incapable of forming compounds (e.g., salts) with zinc, or 2. A physiologically acceptable injectable (e.g., intramuscularly injectable and / or, more preferably, subcutaneously injectable) buffer system containing a source of counterion capable of forming a compound (e.g., a salt) with zinc, whose counterion is more soluble in the aqueous medium than zinc hydroxide (Zn(OH)).

[0172] The inventors have found that when coated particles are present in an aqueous medium with a coating (and / or at least the outermost layer of coating material) comprising zinc oxide (e.g., a zinc content of at least about 1 mg, e.g., at least about 5 mg (e.g., at least about 10 mg) of zinc in the coated particle per mL of aqueous carrier) at medium to high concentrations (e.g., between at least about 5 mg (e.g., at least about 10 mg) of at least one GLP1RA per mL of carrier, e.g., about 25 mg to about 200 mg of active agent per mL of carrier, e.g., about 30 mg / mL to about 150 mg / mL, e.g., about 40 mg / mL to about 100 mg / mL, e.g., about 50 mg / mL), viscous aggregates and / or hydrogels may form, which have a tendency to clog needles when the formulation is intended to be injected, for example, subcutaneously or intramuscularly.

[0173] The inventors have discovered that the unexpected formation of aggregates and / or hydrogels is caused by the reaction of zinc in the coating with water to form zinc hydroxide, resulting in an unstable and non-injectable suspension.

[0174] The present inventors have discovered that this unexpected problem can be solved by the addition of a compound that reacts with zinc in competition with water. Such a compound can thus react with zinc, thereby reducing and / or substantially preventing (e.g., up to about 75%, e.g., about 80%, including up to about 90%, e.g., about 95%), even up to about 99%, the reaction of zinc with water to form zinc hydroxide.

[0175] Such compounds may, for example, provide a source of counterions for the aqueous carrier system, which, when included (e.g., dissolved) in the latter, may form compounds (e.g., salts) with zinc that serve to prevent the above-mentioned reaction between free zinc and water. Such counterions may achieve this formation, for example, by complexing with zinc in some way, and / or by being essentially insoluble in water, and / or by precipitating from aqueous solution, and this competitive reaction may occur at or near the surface of the coated particles. Whatever the mechanism, in competing with water to react with zinc, further reaction between zinc and water (and thus gelation) is reduced and / or prevented.

[0176] The term "essentially insoluble" in an aqueous medium (e.g., pure water) includes zinc compounds (e.g., salts) that are sparingly soluble in such a medium, such as those having a solubility of less than about 33.3 mg / mL, e.g., less than about 25 mg / mL (less than about 20 mg / mL), in particular less than about 10 mg / mL, e.g., less than about 5 mg / mL, at least less than about 1 mg / mL, and less than about 0.1 mg / mL at atmospheric pressure (e.g., about 1 bar), room temperature (e.g., about 21°C) and neutral pH (e.g., a pH value of about 5 to about 9, e.g., about 6 to about 8.5, e.g., about 7 to about 8 (e.g., about 7.4)).

[0177] The solubility of zinc-related compounds (e.g., salts) in aqueous media (e.g., pure water) is preferably less than the solubility of zinc hydroxide (Zn(OH)2) (e.g., at least about 10%, e.g., at least about 5%) at any given temperature, pressure, and pH.

[0178] In this regard, suitable counterions having the aforementioned properties include aspartate, tartrate, maleate, fumarate, malate, benzoate, and preferably phosphate counterions. Suitable sources of such counterions include aspartic acid, aspartate (e.g., sodium aspartate) and hydrates thereof, and mixtures of these components; tartaric acid, tartrate (sodium tartrate) and hydrates thereof (e.g., dibasic hydrate), and mixtures of these components; maleic acid, maleate (e.g., sodium maleate) and hydrates thereof, and mixtures of these components; fumaric acid, fumarate (e.g., monosodium fumarate) and hydrates thereof, and mixtures of these components. Included are materials capable of forming buffer solutions based on aspartic acid, tartaric acid, maleic acid, fumaric acid, malic acid, benzoic acid and / or phosphate, such as mixtures thereof, malic acid, malic acid salts and hydrates thereof, and mixtures of these components, benzoic acid, benzoic acid salts (e.g., sodium benzoate) and hydrates thereof, and mixtures of these components, and more particularly phosphate buffers (such as phosphoric acid, disodium hydrogen phosphate dihydrate, sodium acid phosphate, sodium dihydrogen phosphate monohydrate, and combinations thereof).

[0179] Alternative sources of phosphate counterions include salts such as sodium phosphate, potassium phosphate, and calcium phosphate. Sources may also include organic phosphates (e.g., glycerol phosphate, sodium glycerophosphate, and potassium glycerophosphate), as well as sources of pyrophosphate and polyphosphate. Alternative sources of tartrate counterions include potassium tartrate, diethyl tartrate, and disodium tartrate. Alternative sources of benzoate counterions include sodium benzoate, benzyl benzoate, denatonium benzoate, and potassium benzoate.

[0180] A suitable concentration of phosphate counterions in the aqueous medium of the formulations of the present invention can be in the range of about 1 mM, e.g., about 2 mM to about 50 mM (including about 40 mM), e.g., about 3 mM to about 35 mM, e.g., about 4 mM (e.g., about 5 mM) to about 30 mM (e.g., about 25 mM, including about 20 mM, about 15 mM, and about 10 mM). (For example, the recommended upper concentration limit for phosphate buffer in subcutaneously injectable compositions is 10 mM; see Usach et al., Adv. Ther., 36, 2986 (2019)).)

[0181] While the above-mentioned counterion sources capable of forming zinc compounds (e.g., salts) may prevent the gelation problems described herein, the inventors have unexpectedly found that this can cause changes in the pH of the resulting formulation (particularly over time, e.g., during storage), which is believed to result from the reaction of the counterion(s) (e.g., phosphate ions) with zinc, which depletes the buffering capacity of the associated counterion source (e.g., buffer(s)).

[0182] The inventors have discovered that this problem is a counterion source capable of forming a salt with zinc that is more soluble in aqueous media than zinc hydroxide (Zn(OH)2), or more preferably, We have found that this problem can be solved by including a physiologically acceptable injectable (e.g., intramuscularly injectable, or more preferably, subcutaneously injectable) buffer system that is essentially incapable of forming a salt with zinc.

[0183] The inventors have found that the presence of such buffering agents, when combined with other counterion sources described above, counteracts the above-mentioned effects of depleting the buffering capacity of the latter and thus serves to maintain a constant (e.g., physiologically acceptable) pH within the formulations of the invention. By "maintaining a constant pH" within the formulations of the invention, the inventors have found that there is a pH variation of less than ±20%, for example, less than ±10%, including less than ±5%, during storage when compared to the pH measured immediately after preparation of the formulations of the invention.

[0184] Buffers containing a counterion source capable of forming a salt with zinc that is more soluble in aqueous media than zinc hydroxide include citrate buffers (e.g., citric acid, trisodium citrate dihydrate, and combinations thereof), acetate buffers (e.g., acetic acid, sodium acetate, and combinations thereof), lactate buffers (e.g., lactic acid, magnesium lactate, and combinations thereof), gluconate buffers (e.g., gluconic acid, sodium gluconate, and combinations thereof), glutamate buffers (e.g., glutamic acid, monosodium glutamate, and combinations thereof), succinate buffers (e.g., , succinic acid, sodium succinate, and combinations thereof), α-ketoglutaric acid buffer (α-ketoglutaric acid, α-ketoglutarate, and combinations thereof), ascorbic acid buffer (e.g., ascorbic acid, sodium ascorbate, and combinations thereof), bicarbonate buffer (e.g., carbonic acid, sodium bicarbonate, and combinations thereof), ammonium buffer (e.g., ammonium chloride, ammonium hydroxide, and combinations thereof), glycine buffer (e.g., glycine, sodium glycinate, and combinations thereof), or a combination of any of the above.

[0185] Buffers that are essentially incapable of forming salts with zinc include histidine, diethanolamine (e.g., diethanolamine, magnesium chloride hexahydrate, and combinations thereof), or, most preferably, tromethamine ("Tris" or "Trizma") buffers.

[0186] All of the aforementioned buffering agents can be used alone or in combination with standard inorganic acids and bases, such as hydrochloric acid and sodium hydroxide, which can be used to adjust the pH. A preferred pH for the formulations of the present invention can be in the range of about pH 3 to about pH 10, for example, about pH 4 to pH 9 (including pH 5 to about pH 8).

[0187] A suitable concentration of such a buffer (e.g., tromethamine buffer) is within the range of about 0.1 mM (e.g., about 5 mM (including about 10 mM)) to about 200 mM, for example, about 25 mM to about 175 mM, for example, between about 50 mM and about 150 mM (including about 75 mM to about 125 mM (e.g., about 100 mM)).

[0188] Formulations of the present invention may also include a carrier system that is a mixture of one or more oils and an aqueous system, for example an emulsion such as an oil-in-water emulsion or a water-in-oil emulsion.

[0189] In (especially oil-based) carriers, suitable dispersing or wetting agents (for example Tween such as Tween 80), and / or suspending agents may be used.

[0190] Formulations of the present invention may also include a carrier system that is a mixture of one or more oils and an aqueous system, for example an emulsion such as an oil-in-water emulsion or a water-in-oil emulsion.

[0191] In (especially oil-based) carriers, suitable dispersing or wetting agents (for example Tween such as Tween 80), and / or suspending agents may be used.

[0192] Thus, the formulations can be stored under normal storage conditions and maintain their physical and / or chemical integrity. The phrase "maintaining physical and chemical integrity" essentially means chemical stability and physical stability.

[0193] By "chemical stability" we include that any formulation of the present invention can be stored under normal storage conditions (with or without appropriate pharmaceutical packaging) with little chemical (including stereochemical) degradation or degradation of any active ingredient (in particular any GLP1RA) and / or inactive excipients, and / or change in pH as described above.

[0194] "Physical stability" includes that the formulations of the present invention can be stored (with or without appropriate pharmaceutical packaging) under normal storage conditions with only a small degree of physical transformation (e.g., precipitation as described above) or change in the properties and / or integrity of the coated particles, e.g., the coating itself or the active ingredient (including dissolution, solvation, solid-state phase transitions, etc.).

[0195] "Normal storage conditions" for the formulations of the present invention include, for example, temperatures of about -50°C to about +80°C (preferably about -25°C to about +75°C, such as about 50°C) for an extended period of time (i.e., about 12 months or more, e.g., about 6 months), and / or pressures of about 0.1 to about 2 bar (preferably atmospheric pressure), and / or exposure to ultraviolet / visible light of about 460 lux, and / or relative humidity of about 5 to about 95% (preferably about 10 to about 40%).

[0196] Under such conditions, the formulations of the present invention may be found to be chemically and / or physically degraded / decomposed, optionally to less than about 15%, more preferably less than about 10%, and especially less than about 5%. Those skilled in the art will appreciate that the above upper and lower limits of temperature and pressure represent the extremes of normal storage conditions, and that certain combinations of these extremes will not be experienced during normal storage (e.g., a temperature of 50°C and a pressure of 0.1 bar).

[0197] The formulations of the present invention may comprise from about 1% to about 99% by weight, such as from about 10% (such as about 20% by weight, e.g., about 50% by weight) to about 90% by weight of coated particles, with the remainder made up of carrier systems and / or other pharmaceutically acceptable excipients.

[0198] The formulations of the present invention may also include compositions in the form of a liquid, sol, or gel that can be administered through a surgical administration device, such as a needle, catheter, etc., to form a depot formulation.

[0199] In any event, the preparation of suitable formulations can be accomplished non-inventively by those skilled in the art using routine techniques. Thus, the formulations of the present invention and dosage forms containing same can be formulated with conventional pharmaceutical additives and / or excipients used in the art for the preparation of pharmaceutical formulations, and then incorporated into various types of pharmaceutical preparations and / or dosage forms using standard techniques (see, e.g., Lachman et al., "The Theory and Practice of Industrial Pharmacy," Lea & Febiger, 2003). rd edition (1986), 'Remington: The Science and Practice of Pharmacy', Troy (ed.), University of the Sciences in Philadelphia, 21 st edition (2006), and / or 'Aulton's Pharmaceutics: The Design and Manufacture of Medicines', Aulton and Taylor (eds.), Elsevier, 4 th edition, 2013), and documents cited therein, the relevant disclosures of all of which are incorporated herein by reference).

[0200] According to a further aspect of the present invention, there is provided a process for preparing a formulation of the present invention, comprising mixing coated particles, e.g., as described herein, together with an aqueous carrier system, e.g., as described herein.

[0201] For intramuscular or, particularly, subcutaneous injection, the formulations of the present invention may be presented in the form of a sterile injectable dosage form that can be administered via a surgical administration device (e.g., a syringe with an injection needle, catheter, etc.) to form a depot preparation.

[0202] Particularly preferred administration devices include auto-injectors, which are known to those skilled in the art as a type of medical device specifically designed for a patient to self-administer a predetermined dose of a drug, and are typically single-use, disposable, spring-loaded injectors, including any of those currently known in the art for administering GLP1RAs.

[0203] Thus, an injectable dosage form is provided comprising a formulation of the invention, the formulation being contained in a reservoir connected to and / or associated with an injection means (e.g., a syringe equipped with a needle or the like for injection).

[0204] Alternatively, the formulations of the present invention may be stored before being loaded into a suitable injectable dosage device (e.g., a syringe equipped with a needle for injection) or may be prepared immediately before being loaded into such a dosage device.

[0205] Thus, the sterile injectable dosage form may comprise a receptacle or reservoir in communication with the injection means which may be pre-loaded with the formulation of the invention, which may be loaded prior to use, or may comprise one or more reservoirs in which the formulation of the invention and the coated particles of the aqueous carrier system are separately contained and mixing occurs before and / or during injection.

[0206] therefore, (a) coated particles of the formulation of the present invention; (b) a carrier system for the formulation of the present invention; and As well as, there is further provided a kit of parts comprising coated particles of the formulation of the invention together with instructions to the end user for mixing the particles with a carrier system according to the invention.

[0207] Further provided are pre-loaded injectable dosage forms as described above, but modified by including at least two chambers, in one of which the coated particles of the formulation of the invention are disposed and in the other chamber the aqueous carrier system of the formulation of the invention is disposed, and upon mixing, a suspension or otherwise is formed before and / or during injection.

[0208] The formulations of the present invention can be used in human medicine.The formulations of the present invention are particularly useful in any indication for which GLP1RA is approved for use or is otherwise known to be useful.In particular, the formulations of the present invention are useful for treating metabolic conditions or disorders.

[0209] The term "metabolic condition or disorder" is understood to include endocrine diseases and / or disorders, and thus disorders of the glands of the endocrine system (including the adrenal glands, thyroid gland, pituitary gland, and especially the pancreas), and therefore includes all types of diabetes, such as hyperglycemia, type 1 diabetes, type 2 diabetes, impaired glucose tolerance, non-insulin-dependent diabetes, mature onset diabetes of the young, gestational diabetes, and conditions related to weight and / or overeating, such as obesity, which can be induced by force of habit or by drugs, such as steroids and antipsychotics, and exogenous sources, such as binge eating disorder, bulimia nervosa, etc. This term may also include the medical term "metabolic syndrome."

[0210] A human subject suffering from obesity (obese subject) can have a body mass index (BMI, calculated as a person's weight in kilograms divided by the square of the subject's height in meters) of at least about 25, e.g., at least about 27 (including at least about 30), e.g., in the range of about 30 to about 40 (e.g., about 35), or greater than 40.

[0211] Preferred metabolic conditions include type 2 diabetes and obesity, which, as alluded to above, are often considered comorbidities. In addition to diabetes (e.g., type 2 diabetes), excess weight-related comorbidities that may also be treated with the formulations of the present invention include hypertension, dyslipidemia, high cholesterol, and obstructive sleep apnea.

[0212] The formulations of the present invention are indicated in the therapeutic, palliative, and / or diagnostic treatment, as well as the prophylactic treatment (including preventing and / or arresting the deterioration and / or worsening of the condition), of any of the above metabolic disorders or conditions.

[0213] This may include slowing or preventing the progression of diabetic disease, such as the progression of type 2 diabetes, slowing the progression of impaired glucose tolerance (IGT) to insulin-requiring type 2 diabetes, and / or slowing the progression of non-insulin-requiring type 2 diabetes to insulin-requiring type 2 diabetes. It may also include reducing food intake, reducing weight, suppressing appetite, inducing satiety, reducing gastric motility and / or slowing gastric emptying.

[0214] The formulations of the invention may also be useful in the treatment and / or prevention (which, as noted above, may include arresting deterioration and / or worsening) of conditions, disorders or diseases that may be the result of a metabolic condition or disorder, including high blood pressure and more serious adverse cardiovascular events such as myocardial infarction, stroke, angina, heart failure and other cardiovascular disorders; non-alcoholic fatty liver disease such as non-alcoholic fatty liver and non-alcoholic steatohepatitis; other neurodegenerative conditions such as Alzheimer's disease and Parkinson's disease, and other conditions including nerve damage, skin disorders such as chronic diabetic ulcers, diabetic ketoacidosis, eye conditions such as retinopathy and / or kidney disorders (e.g., chronic renal failure), e.g., nephropathy, gum disease and / or mouth problems, cancer and / or sexual dysfunction.

[0215] In treating any of the above conditions, GLP1RAs may be combined with other treatments known to be useful in treating related conditions.

[0216] This includes type 2 diabetes receptor analogs such as GIP, glucagon, sodium glucose cotransporter, amylin and oxyntomodulin / peptide YY, as well as known diabetes therapeutic agents such as insulin, biguanides (e.g., buformin, phenformin, especially metformin), sulfonylureas (e.g., tolbutamide, acetohexamide, tolazamide, chlorpropamide, glipizide, glibenclamide, glimepiride, gliclazide, glyclopyramide and gliquidone), alpha-glucosidase inhibitors (e.g., miglitol, voglibose and acarbose), and the like. These include steroids, including but not limited to steroids, prandial glucose regulators (e.g., repaglinide and nateglinide), peroxisome activated receptor gamma (PPAR) inhibitors or glitazones (e.g., rosiglitazone, troglitazone, or pioglitazone), dipeptidyl peptide-4 (DPP-4) inhibitors (gliptins, e.g., vildagliptin, sitagliptin, saxagliptin, linagliptin, septagliptin, teneligliptin, gemigliptin, and alogliptin), and / or SGLT2 inhibitors (e.g., dapagliflozin, canagliflozin, empagliflozin, and remogliflozin).

[0217] This also includes known anti-obesity agents such as orlistat, cetilistat, loraxerin, sibutramine, rimonabant, metformin, berberine, forskolin, and combinations such as amylin / pramlinatide, phentermine / topiramate, naltrexone / bupropion, superabsorbent hydrogels (e.g., Gelesis 100) and / or statins.

[0218] The formulations of the present invention (with or without the above-mentioned combination therapies) may also be used in conjunction with glucometers and continuous glucose monitors, and / or may involve more invasive techniques such as pancreatic islet cell transplantation or weight loss surgery.

[0219] Furthermore, as described below, it has been found that injection of the formulations of the present invention can cause a mild inflammatory response, which can be mitigated by co-administering a suitable anti-inflammatory agent with the injection.

[0220] Suitable anti-inflammatory agents that may be used in this regard include butylpyrazolidines (such as phenylbutazone, mofebutazone, oxyphenbutazone, clofezone, kebuzone, and suxibuzone), acetic acid derivatives and related substances (such as indomethacin, sulindac, tolmetin, zomepirac, diclofenac, alclofenac, bumadizone, etodolac, lonazolac, fentiazac, acemetacin, difenpyramide, oxametacin, proglumetacin, ketorolac, aceclofenac, and bufexamac), oxicams (such as piroxicam, tenoxicam, dromedazone, thiazolinone ... oxicam, lornoxicam, meloxicam), propionic acid derivatives (such as ibuprofen, naproxen, ketoprofen, fenoprofen, fenbufen, benoxaprofen, suprofen, pirprofen, flurbiprofen, indoprofen, tiaprofenic acid, oxaprozin, ibuproxam, dexibuprofen, flunoxaprofen, alminoprofen, dexketoprofen, vedaprofen, carprofen, and tepoxalin), fenamates (such as mefenamic acid, tolfenamic acid, flufenamic acid, meclofenamic acid, and and flunixin), coxibs (celecoxib, rofecoxib, valdecoxib, parecoxib, etoricoxib, lumiracoxib, firocoxib, robenacoxib, mavacoxib, and cimicoxib), other nonsteroidal anti-inflammatory drugs (nabumetone, niflumic acid, azapropazone, glucosamine, benzydamine, glycosaminoglycan polysulfate, proquazone, orgotein, nimesulide, feprazone, diacerein, morniflumate, tenidap, oxaceprol, chondroitin sulfate, pentosan polysulfate, and aminopropion nitriles, etc.), corticosteroids (11-dehydrocorticosterone, 11-deoxycorticosterone, 11-deoxycortisol, 11-ketoprogesterone, 11β-hydroxypregnenolone, 11β-hydroxyprogesterone, 11β,17α,21-trihydroxypregnenolone, 17α,21-dihydroxypregnenolone, 17α-hydroxypregnenolone, 17α-hydroxyprogesterone, 18-hydroxy-11-deoxycorticosterone, 18-hydroxycorticosterone, 18-hydroxyprogesterone,21-deoxycortisol, 21-deoxycortisone, 21-hydroxypregnenolone (prevedilone), aldosterone, corticosterone (17-deoxycortisol), cortisol (hydrocortisone), cortisone, pregnenolone, progesterone, flugestone (flurogestone), fluorometholone, medrysone (hydroxymethylprogesterone), prevedilone acetate (21-acetoxypregnenolone), chloroprednisone, cloprednol, difluprednate, fludrocortisone, fluocinolone, fluoxetine Perolone, fluprednisolone, loteprednol, methylprednisolone, prednicarbate, prednisolone, prednisone, tixocortol, triamcinolone, alclometasone, beclomethasone, betamethasone, clobetasol, clobetasone, clocortolone, desoximetasone, dexamethasone, diflorasone, difluocortolone, fluchlorolone, flumethasone, fluocortin, fluocortolone, fluprednidene, fluticasone, fluticasone furoate, halometasone, meprednisone, mometasone, mometasone furoate acetaminophen, paramethasone, prednylidene, rimexolone, urobetasol (halobetasol), amcinonide, budesonide, ciclesonide, deflazacort, desonide, formocortal fluchloronone acetonide (flucloronide), fludroxycortide (flurandrenolon, flurandrenolide), flunisolide, fluocinolone acetonide, fluocinonide, halcinonide, and triamcinolone acetonide), quinolines (oxycinchophen, etc.), gold preparations (gold sodium thiomalate, gold sodium thiosulfate, auranofin, aurothiophen glucose, and aurotioprol, etc.), penicillamine and similar drugs (such as bucillamine), antihistamines (acrivastine, alimemazine, antazoline, astemizole, azatadine, azelastine, bamipine, bilastine, bromdiphenhydramine, brompheniramine, buclizine, cetirizine, cinnarizine, cyclizine, cyproheptadine, deptropine, desloratadine, dexbrompheniramine, dexchlorpheniramine, diphenylpyraline, dimenhydrinate, dimethindene, doxylamine, ebastine, epinastine,Anti-inflammatory agents include phenindamine, pheniramine, fexofenadine, histapirrodine, hydroxyethylpromethazine, isothipendyl, carbinoxamine, ketotifen, quifenadine, clemastine, chlorcyclizine, chlorphenamine, chlorphenoxamine, chlorpyramine, levocetirizine, loratadine, mebhydroline, mequitazine, meclozine, mepyramine, methapyrilene, methdilazine, mizolastine, oxatomide, oxomemazine, pimethixene, promethazine, pyrobutamine, rupatadine, sequifenadine, talastine, tenarizine, terfenadine, thiazinam, thiethylperazine, thonzylamine, trimethobenzamide, tripelennamine, triprolidine, and tritocubarine. Combinations of any one or more of the aforementioned anti-inflammatory agents may also be used.

[0221] Preferred anti-inflammatory agents include nonsteroidal anti-inflammatory drugs such as diclofenac, ketoprofen, meloxicam, aceclofenac, flurbiprofen, parecoxib, ketoralac tromethamine, or indomethacin.

[0222] The subject may receive (or may already receive) one or more of the aforementioned co-therapeutic and / or anti-inflammatory agents separate from the formulations of the present invention, meaning that they receive a prescribed dose of one or more of these other therapeutic agents before, in addition to, and / or following treatment with the formulations of the present invention.

[0223] When GLP1RA / its salts are "used in combination" with such other therapeutic agents, the active ingredients may be administered together in the same formulation or separately (simultaneously or sequentially) in different formulations (hereinafter referred to as a "combination product").

[0224] Such combination products provide for administration of a GLP1RA in conjunction with another therapeutic agent and therefore may be presented as separate formulations, at least one of which is a formulation of the present invention and at least one of which contains the other therapeutic agent in a separate formulation, or may be presented (i.e., formulated) as a combined preparation (i.e., may be presented as a single formulation containing a GLP1RA / salt and the other therapeutic agent).

[0225] In this regard, the other therapeutic agent may be presented together with the GLP1RA in an appropriate dose in one or more of the cores forming part of the formulation of the present invention described above, or may be formulated using the same or similar processes for coating as those described above for the GLP1RA, which may allow release of the other therapeutic agent over the same time scale or over a different time scale.

[0226] Thus, there is further provided a pharmaceutical formulation of the invention which further comprises a therapeutic agent useful in the treatment of a metabolic condition such as type 2 diabetes or obesity as defined above, and / or an anti-inflammatory agent.

[0227] Pharmaceutical formulations of the present invention that may be mentioned include those that are free of anti-inflammatory agents and / or are essentially free of the above-mentioned anti-inflammatory agents. By "essentially free of anti-inflammatory agents," we mean that the formulation contains less than about 1% by weight (e.g., less than about 0.5% by weight, such as less than about 0.1% by weight or less than about 0.01% by weight) of one or more of the aforementioned anti-inflammatory agents.

[0228] In such formulations of the invention, the additional therapeutic agent is (1) formulating a GLP1RA into a solid core of a formulation of the present invention (which formulation is hereinafter referred to as a "combination core preparation"); (2) It may be included by dissolving and / or suspending it within the aqueous carrier system of the formulation of the present invention (which formulation is hereinafter referred to as the "combination preparation").

[0229] In embodiment (2) above, the other therapeutic agent may be presented in the formulation of the present invention in any form separate from the GLP1RA-containing core, for example, by directly dissolving or suspending the active ingredient in the aqueous medium of the formulation of the present invention, or by presenting it in a form whose release can also be controlled after injection, similar to GLP1RA.

[0230] The latter option may be achieved, for example, by providing the other therapeutic agent in the form of additional particles suspended in the aqueous carrier system of the formulation of the present invention, the additional particles having an average diameter by weight, number, or volume that is from about 10 nm to about 700 μm, and comprising a core containing a therapeutic agent useful for treating a metabolic disorder as defined herein and / or an anti-inflammatory agent, the core being at least partially coated with one or more coating materials described above (this formulation is hereinafter referred to as a "combination suspension").

[0231] moreover, (A) a pharmaceutical formulation of the present invention; (B) a pharmaceutical formulation comprising a therapeutic agent useful for treating a metabolic disorder as defined herein and / or an anti-inflammatory agent, There is provided a pharmaceutical formulation of the invention in the form of a kit of parts, in which components (A) and (B) are each provided in a form suitable for administration in conjunction with the other.

[0232] Although component (B) of the kit of parts presented above may differ in its chemical composition and / or physical form from component (A) (i.e., the formulation of the present invention), it may also be in a form that is essentially the same as or at least similar to the GLP1RA-containing formulation of the present invention, i.e., in the form of a plurality of particles suspended in a (e.g., aqueous) carrier system, said particles being (a) have an average diameter based on weight, number, or volume of about 10 nm to about 700 μm; (b) A solid core containing another therapeutic agent, which is at least partially coated with one or more coatings of (eg, inorganic) materials.

[0233] In addition, in such a preferred kit-of-parts, and in the combination suspension presented under embodiment (2) above, the coated cores comprising the other therapeutic agents may differ in terms of their chemical composition(s) and / or physical form(s), but the inorganic material coating used is preferably the same as or similar to the coating used in the GLP1RA-containing formulation of the present invention, meaning that the other therapeutic agents are coated with one or more inorganic coatings as described herein, e.g., one or more inorganic coating materials comprising one or more metal-containing or metalloid-containing compounds, e.g., metals, or metalloids, oxides, e.g., iron oxide, titanium dioxide, zinc sulfide, more preferably zinc oxide, silicon dioxide and / or aluminum oxide, which coating materials may essentially constitute (individually or collectively) (e.g., more than about 80%, e.g., about 90%, e.g., about 95%, e.g., about 98%) of such oxides, and more particularly, the inorganic coating is (i) zinc oxide; (ii) with one or more other metal and / or metalloid oxides, The atomic ratio ((i):(ii)) is at least about 1:6 and not more than about 6:1.

[0234] Preferably, the atomic ratio ((i):(ii)) is at least about 1:1 and not more than about 6:1.

[0235] In any event, for the avoidance of doubt, all aspects, including preferred aspects, of the GLP1RA-containing formulations of the invention disclosed and / or claimed herein are equally applicable as aspects and / or preferences of the coated cores comprising one or more of the additional therapeutic agents described above, and for the avoidance of doubt, such aspects, preferences and features, singly or in combination, are hereby incorporated by reference into this specification.

[0236] Thus, all combination products, including the above combination core preparations, combination suspensions and kits of parts, may be used in any indication in human medicine, particularly where GLP1Ra is approved for use or is otherwise known to be useful, such as in metabolic disorders or conditions as defined above, such as type 2 diabetes and / or obesity.

[0237] In certain instances, such additional therapeutic agents, including some of those useful in treating metabolic disorders or conditions defined above, such as, for example, type 2 diabetes and / or obesity, may be referred to as the "standard of care" in connection with a particular condition. The term "standard of care" is understood by those skilled in the art to include the treatment process that a clinician should and / or would be expected to follow for a particular type of patient, disease, and / or clinical situation. For certain new or poorly understood conditions, the standard of care may change and / or evolve over time.

[0238] According to a further aspect of the present invention there is provided a method of making a kit-of-parts as defined above, the method comprising bringing component (A), as defined above, into association with component (B), as defined above, thus rendering the two components suitable for administration in combination with one another.

[0239] By "associating" two components with each other, we mean that components (A) and (B) of the kit: (i) may be provided as separate formulations (i.e., independently of each other) and then combined for use in conjunction with each other in combination therapy; or (ii) may be packaged and presented together as separate components of a "combination pack" for use in conjunction with each other in combination therapy.

[0240] Thus, a kit-of-parts as defined herein, wherein components (A) and (B) are packaged and presented together as separate components of a combination pack for use in combination with one another in a combination therapy; and (I) one of components (A) and (B) described herein, (II) A kit of parts is provided that includes the component together with instructions for use in conjunction with the other of the two components.

[0241] As mentioned above, the kit of parts described herein may include two or more formulations containing appropriate amounts / doses of GLP1RA / salt, and / or two or more formulations containing appropriate amounts / doses of other therapeutic agents, as mentioned above, to provide for repeated administration.

[0242] In this regard, with respect to the kit of parts described herein, "administration in conjunction with" includes the sequential, separate, and / or simultaneous administration of components (A) and (B) of the kit over the course of treatment of the relevant condition.

[0243] Thus, the term "in conjunction with" includes that one or the other of the two formulations may be administered (optionally repeatedly) before, after, and / or simultaneously with the administration of the other component. When used in this context, the terms "co-administered" and "administered simultaneously with" include when the individual doses of the GLP1RA / salt and the other therapeutic agent are administered within 48 hours (e.g., 24 hours) of each other.

[0244] With respect to any of the above combination products according to the invention, each formulation is administered (or, in the case of a kit-of-parts, the two components are administered together, optionally repeatedly) in a manner that may enable a beneficial effect for the subject over the course of treatment of the condition, i.e., a greater extent than if a formulation comprising a GLP1RA / salt alone (e.g., a formulation of the invention) were administered over the same course of treatment (e.g., repeatedly as described herein) in the absence of the other component.

[0245] The determination of whether a combination product provides a greater beneficial effect with respect to treatment and over the course of treatment depends on the condition being treated and / or its severity, but can be routinely accomplished by one of ordinary skill in the art.

[0246] For example, a physician may first administer a formulation of the invention containing the active ingredient alone to treat a patient with a metabolic disorder or condition, and then find that the person exhibits an inflammatory response (which may be caused by the active ingredient itself and / or any other component of the formulation).

[0247] Next, the doctor will: - Kit-of-parts component (B) as described above, combination core preparations, combination preparations, and / or Combination suspension may be administered, any of which includes an anti-inflammatory agent as described above.

[0248] The other active ingredients / therapeutic agents described above that may be used in combination products according to the present invention may be provided in the form of (e.g., pharmaceutically acceptable) salts, including those salts known in the art and described in the medical literature, e.g., Martindale - The Complete Drug Reference, 38 th Edition, Pharmaceutical Press, London (2014) and the documents cited therein, the relevant disclosures of all of which documents are incorporated herein by reference.

[0249] The amount of other active ingredients / therapeutic agents that may be used in combination products according to the present invention must be sufficient to exert its pharmacological effect.

[0250] Therefore, the dose of such other active ingredients that can be administered to a patient must be sufficient to affect the therapeutic response over a reasonable and / or relevant time frame. Those skilled in the art recognize that the selection of the exact dose and composition and the most suitable delivery regimen is influenced not only by the nature of the other active ingredients, but also by, among other things, the pharmacological properties of the formulation, the route of administration, the nature and severity of the condition being treated, the physical condition and mental acuity of the recipient, and the age, condition, weight, sex, and response of the patient being treated, the stage / severity of the disease, and genetic differences between patients.

[0251] Because the administration of the formulations of the present invention can be continuous or intermittent (eg, by bolus injection), the dosage of such other active ingredients will also be determined by the timing and frequency of administration.

[0252] In any event, a physician or other skilled artisan can routinely determine the actual dosage of any particular additional active ingredient that will be most suitable for an individual patient, and dosages of such relevant additional active ingredients are known in the art and are generally described in the medical literature (e.g., Martindale - The Complete Drug Reference, 38 th Edition, Pharmaceutical Press, London (2014) and documents referenced therein, the relevant disclosures of all of which documents are incorporated herein by reference.

[0253] Use of the formulations of the present invention may be beneficial by reducing any burst effect (e.g., concentration maxima immediately after administration) as described above and / or by reducing the C in the plasma concentration-time profile. 最大 By lowering the dissolution rate of GLP1RAs, it is possible to control their dissolution rate and influence their pharmacokinetic profile.

[0254] The formulations of the present invention may also provide a release and / or pharmacokinetic profile that increases the length of release of the GLP1RA from the formulation.

[0255] These factors not only reduce the frequency or need to administer the formulation to a person suffering from the relevant metabolic condition or disorder, but also give the patient more time off the hook and therefore a better quality of life, as well as the benefits of fewer / less frequent, inconvenient and / or painful injections.

[0256] The formulations of the present invention also have the advantage that by controlling the release of the active ingredient at a steady rate over an extended period of time, they provide a low daily exposure to the associated active ingredient, which is expected to reduce undesirable side effects.

[0257] The formulations and processes described herein may have advantages for the physician and / or patient in treating the associated metabolic condition, such as being more convenient, more effective, less toxic, having a broader spectrum of activity, more potent, causing fewer side effects, and having other useful pharmacological properties than any similar treatment known in the prior art.

[0258] Whenever the word "about" is used herein, for example, in the context of an amount (e.g., number, concentration, dimension (size and / or weight), dose, duration, pharmacokinetic parameter, etc.), relative amount (percentage, weight ratio, size ratio, atomic ratio, aspect ratio, proportion, multiple or fraction, etc.), relative humidity, lux, temperature, or pressure, it will be understood that such variable is an approximation and, therefore, can vary by ±15%, e.g., ±10%, e.g., ±5%, preferably ±2% (e.g., ±1%) from the numerical value specified herein. This is true even when such a number is presented as a percentage to begin with (e.g., "about 15%" can mean ±15% of the number 10, which is anywhere between 8.5% and 11.5%).

[0259] The present invention is illustrated by the following non-limiting examples with reference to the accompanying drawings. [Example]

[0260] Example 1 Coated liraglutide microparticles R&D grade liraglutide (MedChemExpress, New Jersey, US) with a purity of 98.5% and a peptide content of 90.8% was suspended in a solution of 0.1% Span 85 (Sigma-Aldrich, MO, USA) in cyclohexane (Merck, Germany).

[0261] The particle size distribution was determined by laser diffraction (SALD-7500 nano (Shimadzu, Japan), 405 nm laser) as follows: %D(10): 2.0 μm, %D(50): 7.3 μm, %D(90): 23.8 μm.

[0262] The raw materials are mixed with purified water (0.8 to 2 MΩ / cm 2 ) to form a milky white liquid with a liraglutide concentration of 15 wt%, which was then spray-dried in a mini spray dryer (B-290, dehumidifier B-296 equipped with a two-fluid nozzle, BUECHI Labortechnik GmbH, Germany).

[0263] The dispersion liquid was poured into a tube at an inlet temperature of 115°C and a suction speed of 100% (approximately 35 m 3 / h), pump speed 8% (3.8 mL / min), nozzle clean 2, volume flow rate 35 mm (N, approximately 600 L / h), which resulted in an outlet temperature of 73° C. The mass yield was approximated to 77.5%.

[0264] The spray-dried material was assayed using a Nexera UPLC-UV-DAD (Shimadzu, Japan) equipped with a SunShell nC18-WP, 4.6 × 100 mm, 2.6 μm particle size column (Chromanik Technologies Inc., Japan) and was found to have a liraglutide content of 100.1 ± 0.2% (relative) in the spray-dried material.

[0265] The particle sizes of the spray dried material were determined as described above and were as follows: %D(10): 1.7 μm, %D(50): 8.2 μm, %D(90): 21.2 μm.

[0266] The spray-dried liraglutide particles were first coated with three layers of pure aluminum oxide by the following processes a to e: a. The valve on the piping between the pump and the ALD reactor was closed. b. Next, the valve on the trimethylaluminum (TMA) precursor bottle was opened for 1 second, filling the vaporized metal-containing precursor into the ALD reactor for 1 second. c. The valve to the precursor bottle was closed and the chamber was allowed to rest for 30 seconds (soak time) before opening the pump again to ensure the metal-containing precursor vapor reacted with the surface of the drug particles. d. The ALD reactor was then pumped for 9 seconds. e. The above steps a to d were repeated 20 times.

[0267] The chamber was then purged with a continuous flow of nitrogen to remove unreacted reagents and organic gases. Steps (a) through (e) above were then essentially repeated, except that water was used as the second reagent to form a distinct aluminum oxide layer on the surface of the active ingredient microparticles. This was followed by another purge pulse using a continuous flow of nitrogen to remove gaseous water and organic gases.

[0268] The above procedure was repeated twice to form a total of three initial aluminum oxide atomic layers.

[0269] This was followed by applying three layers of zinc oxide by repeating the ALD steps above using diethylzinc (DEZ) and then water as precursors in step (b) above, forming a total of three zinc oxide layers. This was followed by coating with one layer of aluminum oxide using the same precursors as described above in (b). This provided a mixed oxide layer with an atomic ratio of 1:3 (Al2O3:ZnO) for a total of four atomic layers.

[0270] This was repeated a total of nine times to form a mixed oxide (1:3 Al2O3:ZnO) layer with a total of 40 atomic layers (30 layers of ZnO and 10 layers of Al2O3).

[0271] The powder was then removed from the ALD reactor and deagglomerated using a sonic sifter (Tsutsui Sonic Agitated Sifting Machine SW-20AT) with a sieve of 20 μm mesh size.

[0272] The resulting deagglomerated powder was reloaded into the ALD reactor, and the same procedure was followed to coat the particles with mixed oxide (1:3 AlO:ZnO) layers for a total of 40 atomic layers. The deagglomeration and coating steps were repeated twice to produce a sample with a pure aluminum oxide undercoat and four separate coatings of mixed oxide with a 1:3 AlO:ZnO atomic ratio.

[0273] The resulting particles were then divided into two batches: for the first batch, an additional outer layer of pure aluminum oxide was applied using the same pure aluminum oxide coating procedure as above.

[0274] For the second batch, the above procedure to produce 40 layers of mixed Al2O3:ZnO (1:3) oxide was repeated twice with deagglomeration to provide two additional separate coatings of mixed oxide, then finished with an additional outer layer of pure aluminum oxide as above.

[0275] Thus, batch 1 contained 3 (inner) layers of Al2O3, 4 × 40 layers of 1:3 Al2O3:ZnO mixed oxide and 3 (outer) layers of Al2O3, and batch 2 contained 3 (inner) layers of Al2O3, 6 × 40 layers of 1:3 Al2O3:ZnO mixed oxide and 3 (outer) layers of Al2O3.

[0276] The particle size of the batches was determined by suspending the coated particles in a solution of 0.1% Span 85 (Sigma-Aldrich, MO, USA) in heptane (Merck, Germany). The particle size distribution measured by laser diffraction was Batch 1 %D(10): 3.5±0.3μm, %D(50): 10.7±0.6μm, %D(90): 23.6±0.6μm, Batch 2 %D(10): 3.2±0.2 μm, %D(50): 9.7±0.6 μm, %D(90): 22.3±0.8 μm.

[0277] The drug loading of the coated batches was determined by etching the samples in a solution of 43% acetonitrile (Spectrascan, Scotland), 45% water (PanReac, Spain), and 12% (v / v) phosphoric acid (Merck, Germany) to dissolve the coating, followed by dilution to 160 μg / mL with 10% acetonitrile + 0.1% trifluoroacetic acid in water (Merck, Germany) (mobile phase A) on a column Kinetex C18 PS, 150 × 4.6 mm, 2.6 μm particle size (Phenomenex Ltd., CA, USA), mobile phase B: 80% acetonitrile in water + 0.1% trifluoroacetic acid, injection volume 10 μL, autosampler temperature 15°C, oven temperature 25°C, flow rate 1.0 mL / min gradient elution, UV absorbance detection at 220 nm. Single-point calibration was used for the assay. The retention time of liraglutide was 6.8 min. The reference material by assay was 99.4% total liraglutide, with a purity of 99.1% by HPLC (Bachem AG, Switzerland), as determined by injection into a quantification HPLC system (Prominence-I HPLC-UV-DAD, Shimadzu, Japan).

[0278] Batch 1 had a drug loading of 61.9±0.2%, and batch 2 had a drug loading of 53.1±0.4%. The purity of the coated liraglutide in both cases was measured as 98.9%. Therefore, no significant decrease in the purity of the coated liraglutide was observed, indicating that it tolerates the ALD process without degradation.

[0279] To determine coating integrity, samples from both batches were suspended in dimethyl sulfoxide (Rathburn, UK) at a concentration of 0.4 mg liraglutide per mL of solvent and rotated on an overhead stirrer for up to 72 hours. Intermittent samples were taken, centrifuged at 6000 rpm for 7 minutes (EBA20, Hettich, Germany), and the supernatant was diluted with mobile phase A (as above) before being injected into the HPLC system for quantification. The results are shown in Figure 1 (Batch 1 (circles), Batch 2 (squares)) and indicate minimal defects in the coating, so any burst release is expected to be low.

[0280] Sample material from batch 1 and sample material from uncoated spray-dried material were subjected to radiation exposure at doses in the ranges of 20.4-20.6 kGy and 23.5-25.3 kGy. 60 The coated materials were subjected to sterilization levels of gamma irradiation from a Co source (Ionisos Baltics OUE, Alliku, Estonia). The drug loading of the coated materials was determined using the same method as above and compared to uncoated material. The absolute reduction in drug loading of Batch 1 material gamma-irradiated at approximately 21 kGy was less than 5% units, indicating that this method can be used to sufficiently non-destructively terminally sterilize this material. The results are shown in Figure 2 (error bars indicate standard deviation (n = 3); ns indicates lack of significant difference (p > 0.05) as determined by 2-way ANOVA with Dunn-Sidak correction). The purity of liraglutide in the processed materials was determined using the same method as above and confirmed the results from the drug loading assay in all samples at greater than 95%.

[0281] Example 2 Formulation I of the present invention The coated liraglutide particles (Batch 1 and Batch 2 from Example 1 above) are suspended in a suitable aqueous carrier, such as 5-30 mM phosphate buffered saline and 100 mM tromethamine buffer, made up to volume with water for injection to provide a suspension having a concentration of about 100 mg of liraglutide per mL of aqueous carrier. The pH of the resulting formulation is measured as about 7.4.

[0282] Example 3 Formulation II of the Invention Two suspensions of liraglutide-coated microparticles (Batch 1 and Batch 2, prepared according to the process described in Example 1 above) were prepared by adding an appropriate volume of Hyonate vet. (a 1 wt% hyaluronic acid solution in 1 mM phosphate buffer, pH 7.4) to test article vials to achieve a liraglutide concentration equivalent to 10 mg / mL. The vials were then tapped at least 10 times to remove any material that may have settled to the bottom of the test vial. The formulations were then vortexed for approximately 60 seconds to ensure a uniform suspension.

[0283] Example 4 In vivo studies Sixteen male Sprague Dawley rats weighing approximately 300 g on the day of dosing were supplied by Charles River Labs (UK). The animals were randomly divided into four animals per group.

[0284] The hair in the intended administration area was clipped prior to injection, and the injection site was marked. The suspension (prepared as described in Example 3 above) was drawn into a 1 mL syringe and a single subcutaneous injection (approximately 0.08 mL (Batch 1 and Batch 2, Groups 1 and 3, as identified in Table 1 below) and 0.17 mL (Batch 2, Group 2, as identified in Table 1 below) was administered subcutaneously through a 23G needle into the flank of each rat. Comparator liraglutide solution (Victoza, Novo Nordisk, Denmark) was diluted to 0.6 mg / mL with normal injectable saline (Group 4, as identified in Table 1 below), drawn into a 1 mL syringe, and administered subcutaneously through a 23G needle into the flank of each rat. Administration occurred within 30 minutes of formulation preparation.

[0285] The dosing site on each animal was marked after dosing and kept hairless throughout the study. Observations of the dosing sites were made 1 hour and 24 hours after dosing, and then daily until the final sampling point. [Table 1]

[0286] Blood samples (approximately 0.2 mL) were collected from the jugular vein into KEDTA (dipotassium ethylenediaminetetraacetic acid) tubes at 1, 3, 6, 12, 24, 48, 72, 120, 168, 251, 384, 480, 576, and 672 hours after dosing for Groups 1-3, and at 1, 2, 3, 6, 9, 12, 24, and 48 hours after subcutaneous dosing for Group 4. Actual sampling times were recorded. Plasma was separated by centrifugation (1500 g, 10 minutes at 4°C) as soon as practical after blood sampling and stored at -80°C until analysis.

[0287] After study completion, all plasma samples were shipped for analysis deep frozen on dry ice. Animals were sacrificed on the final day of the study.

[0288] The plasma concentration of liraglutide was determined by LC-MS / MS. Study samples were prepared by pipetting 35 μL of rat plasma into a 96-well plate and adding 75 μL of internal standard working solution using an EVO-2 liquid handling robot (Tecan, Austria). The 96-well plate was then shaken for 15 minutes and centrifuged. All samples were then injected onto a UPLC-MS / MS system (Waters, MA, USA). Separation was performed using an ACQUITY UPLC Protein BEH C4 column, 300 Å, 1.7 μm, 2.1 mm × 50 mm (Waters, MA, USA) at 60 °C using 0.3% formic acid in water as mobile phase A and acetonitrile as mobile phase B.

[0289] The pharmacokinetic (PK) analysis of liraglutide in plasma was evaluated using non-compartmental analysis (NCA) utilizing the software Phoenix WinNonlin, version 8.3 (Certara, USA). 最大 and t 最大 was derived from the observed plasma concentration data. AUC was estimated by integration of the plasma concentration versus time curve using linear interpolation for increasing plasma levels and logarithmic interpolation for decreasing plasma levels (linear up-log down method). AUC ∞ The measurable plasma concentration (AUC 最後 ) and then extrapolate to infinity using the concentration in the last quantifiable sample and lambda z to find the curve t 1 / 2,z The first-order rate constant associated with the terminal portion of z was calculated by

[0290] result Dose-normalized plasma concentrations of liraglutide over a 4-week period following a single subcutaneous administration of the various formulations are shown in Figure 3, and Figure 4 shows the same plasma concentration profiles over the first 72 hours.

[0291] Plasma pharmacokinetic parameters are also presented below in Table 2 as mean values ​​for groups of 6 rats (standard deviations are shown in brackets). "t 最大 ” is the time to peak concentration in hours "C 最大 " is the maximum concentration found in the assay expressed in ng / mL "t 最後 ” is the time of the last detectable concentration expressed in hours "t 1 / 2,z " is the terminal half-life in hours "AUC ∞ ” is the area under the concentration versus time curve extrapolated to infinity expressed in ng*h / mL "F" is the relative bioavailability expressed as a percentage "C 最大 / D" is the maximum concentration normalized to 1 mg / kg expressed in ng / mL / mg / kg body weight of rat "AUC last / D" is the area under the blood concentration versus time curve up to the last detectable concentration normalized to 1 mg / kg expressed in ng*h / mL / mg / kg body weight in rats "AUC ∞ / D" is the area under the concentration versus time curve to infinity normalized to 1 mg / kg expressed in ng*h / mL / mg / kg body weight in rats · “Fr.Rel. 0-24h " is the fraction released during the first 24 hours of the area under the concentration versus time curve to infinity expressed as a percentage. [Table 2]

[0292] The plasma concentration profiles were similar for Groups 1 to 3, with maximum plasma concentrations (C 最大) was reached within the first 24 hours, then declined rapidly until 72 hours after administration, followed by a slow but steady decline in concentrations over the 28-day study period, compared to Group 4, which showed complete release after 2 days.

[0293] Group 1 had a higher initial drug release compared to the thicker coated formulations of Groups 2 and 3.

[0294] Comparing groups 1-3 with each other, it can be seen that systemic exposure to liraglutide was highly proportional to the administered dose, regardless of coating thickness.

[0295] Groups 1-3 demonstrated a relative bioavailability (F) that was lower than that of liraglutide solution. The trend was proportional to the relative amount of the administered dose released within the first 24 hours, but was not statistically significant.

[0296] C 最大 was higher for liraglutide administered as mixed oxide-coated particles (Groups 1-3) compared with liraglutide solution (Group 4), with approximately one-fifth of the dose released during day 1 for Group 2, one-third for Group 3, and two-fifths for Group 1. When normalized for dose, the difference is approximately one order of magnitude for the thicker coatings (Groups 2 and 3). Also, the remaining area (correlating to unreleased drug) after the last sampling time (672 hours post-dose) was less than 5%.

[0297] Results for Group 1 showed a greater fraction of the dose released on the first day, a higher C 最大 / D and showed a slightly different profile characterized by a shorter duration.

[0298] In summary, Groups 1 to 3 exhibited sustained release profiles distinct from the rapid decline following administration of liraglutide solution. Similarly, favorable plasma concentration-time profiles were observed for all formulations of the present invention, but particularly for Groups 2 and 3.

[0299] Example 5 Coated liraglutide microparticles Coated liraglutide microparticles were prepared using a method similar to that described in Example 1, resulting in two batches corresponding to batch 3 and batch 4.

[0300] The particle sizes of the spray-dried liraglutide particles were determined as described above and were as follows: %D(10): 1.5 μm ± 2.5%, %D(50): 5.2 μm ± 2.9%, %D(90): 11.5 μm ± 2.3%.

[0301] Batch 3 For batch 3, the spray-dried liraglutide particles were first coated with three layers of pure aluminum oxide by the following process: a. The valve on the piping between the pump and the ALD reactor was closed. b. Next, the valve on the trimethylaluminum (TMA) precursor bottle was opened for 1 second, filling the vaporized metal-containing precursor into the ALD reactor for 1 second. c. The valve to the precursor bottle was closed and the chamber was allowed to rest for 30 seconds (soak time) before opening the pump again to ensure the metal-containing precursor vapor reacted with the surface of the drug particles. d. The ALD reactor was then pumped for 9 seconds. e. The above steps a to d were repeated 20 times.

[0302] The chamber was then purged with a continuous flow of nitrogen to remove unreacted reagents and organic gases. Steps (a) through (e) above were then essentially repeated, except that water was used as the second reagent to form a distinct aluminum oxide layer on the surface of the active ingredient microparticles. This was followed by another purge pulse using a continuous flow of nitrogen to remove gaseous water and organic gases.

[0303] The above procedure was repeated twice to form a total of three initial aluminum oxide atomic layers.

[0304] This was followed by applying one layer of zinc oxide by repeating the ALD steps above using diethylzinc (DEZ) and then water as precursors in step (b) above, forming a total of one zinc oxide layer. This was followed by coating with one layer of aluminum oxide using the same precursors as described above in (b). This provided a mixed oxide layer with a 1:1 atomic ratio (Al2O3:ZnO) for a total of four atomic layers.

[0305] This process was repeated 11 times to form a mixed oxide (1:1 Al2O3:ZnO) layer with a total of 25 atomic layers (11 layers of ZnO and 14 layers of Al2O3), including three initial aluminum oxide atomic layers.

[0306] The powder was then removed from the ALD reactor and deagglomerated using a sonic sifter (Tsutsui Sonic Agitated Sifting Machine SW-20AT) with a sieve of 20 μm mesh size.

[0307] The resulting deagglomerated powder was reloaded into the ALD reactor, and the same procedure was followed to coat the particles with three layers of aluminum oxide, followed by a mixed oxide (1:1 AlO:ZnO) layer, for a total of 25 atomic layers. The deagglomeration and coating steps were repeated four times to produce a sample with a pure aluminum oxide undercoat layer and five separate coatings of mixed oxide with a 1:1 AlO:ZnO atomic ratio.

[0308] Thus, batch 3 contained 5 x 25 layers, i.e., 5 x (3 layers of Al2O3 and 22 layers of 1:1 Al2O3:ZnO mixed oxide).

[0309] Batch 4 For batch 4, the spray-dried liraglutide particles were first coated with 10 layers of pure aluminum oxide by the following processes a to e: a. The valve on the piping between the pump and the ALD reactor was closed. b. Next, the valve on the trimethylaluminum (TMA) precursor bottle was opened for 1 second, filling the vaporized metal-containing precursor into the ALD reactor for 1 second. c. The valve to the precursor bottle was closed and the chamber was allowed to rest for 30 seconds (soak time) before opening the pump again to ensure the metal-containing precursor vapor reacted with the surface of the drug particles. d. The ALD reactor was then pumped for 9 seconds. e. The above steps a to d were repeated 20 times.

[0310] The chamber was then purged with a continuous flow of nitrogen to remove unreacted reagents and organic gases. Steps (a) through (e) above were then essentially repeated, except that water was used as the second reagent to form a distinct aluminum oxide layer on the surface of the active ingredient microparticles. This was followed by another purge pulse using a continuous flow of nitrogen to remove gaseous water and organic gases.

[0311] The above procedure was repeated nine times to form a total of 10 initial aluminum oxide atomic layers.

[0312] This was followed by applying three layers of zinc oxide by repeating the ALD steps above using diethylzinc (DEZ) and then water as precursors in step (b) above, forming a total of three zinc oxide layers. This was followed by coating with one layer of aluminum oxide using the same precursors as described above in (b). This provided a mixed oxide layer with an atomic ratio of 1:3 (Al2O3:ZnO) for a total of four atomic layers.

[0313] This process was repeated a total of nine times to form a mixed oxide (1:3 Al2O3:ZnO) layer with a total of 50 atomic layers (30 layers of ZnO and 20 layers of Al2O3), including 10 initial aluminum oxide atomic layers.

[0314] The powder was then removed from the ALD reactor and deagglomerated using a sonic sifter (Tsutsui Sonic Agitated Sifting Machine SW-20AT) with a sieve of 20 μm mesh size.

[0315] The resulting deagglomerated powder was reloaded into the ALD reactor, and the same procedure was followed to coat the particles with 10 layers of pure aluminum oxide, followed by a layer of mixed oxide (1:3 Al2O3:ZnO), for a total of 50 atomic layers. After the final deagglomeration step, the same procedure was followed to coat the particles with 10 layers of pure aluminum oxide, followed by a layer of mixed oxide (1:3 Al2O3:ZnO), and finally, three layers of pure aluminum oxide, for a total of 53 atomic layers. The deagglomeration and coating steps were repeated five times to produce a sample with a primed finish layer of pure aluminum oxide and six separate coatings of mixed oxide with an atomic ratio of 1:3 Al2O3:ZnO.

[0316] Thus, batch 4 contained 6x50 layers, i.e. 6x (10 layers of Al2O3 and 40 layers of 1:3 Al2O3:ZnO mixed oxide), and 3 (outer) layers of Al2O3.

[0317] Batch characterization The particle size of the batches was determined by suspending the coated particles in a solution of 1% Span 85 (Sigma-Aldrich, MO, USA) in heptane (Merck, Germany). The particle size distribution measured by laser diffraction was Batch 3: %D(10): 3.7 μm ± 9.9%, %D(50): 9.3 μm ± 3.9%, %D(90): 21.3 μm ± 5.9%, Batch 4: Undecided.

[0318] The drug loading of the coated batches was determined by etching the samples in a solution (PanReac, Spain) of 2 M phosphoric acid (Merck, Germany) in water to dissolve the coating before diluting to 160 μg / mL. For batch 3, the sample diluent was a mixture of 10% mobile phase B (90% acetonitrile (Spectrascan, Scotland) + 0.1% trifluoroacetic acid (Sigma Aldrich, Germany) in water) and 90% mobile phase A (10% acetonitrile + 0.1% trifluoroacetic acid). For batch 4, the sample diluent was 0.1% trifluoroacetic acid. The sample was injected into a quantification HPLC system (Prominence-I HPLC-UV-DAD, Shimadzu, Japan) using a Kinetex F5 column, 100 Å, 150 × 4.6 mm, 2.6 μm particle size (Phenomenex Ltd., CA, USA), mobile phase A: 10% acetonitrile in water + 0.1% trifluoroacetic acid, mobile phase B: 90% acetonitrile in water + 0.1% trifluoroacetic acid, injection volume 10 μL, autosampler temperature 20 °C, oven temperature 25 °C, gradient elution at a flow rate of 1.0 mL / min, UV absorbance detection at 220 nm. Single-point calibration was used for the assay. The retention time of liraglutide was 6.8 min. The assay reference material was 99.4% total liraglutide, and the purity by HPLC was 99.1% (Bachem AG, Switzerland).

[0319] Batch 3 had a drug loading of 70.5±0.7% and batch 4 had a drug loading of 46.8±0.2%.

[0320] To determine coating integrity, samples from both batches were suspended in dimethyl sulfoxide (Rathburn, UK) at a concentration of 0.4 mg liraglutide per mL of solvent and rotated on an overhead stirrer for 3 hours. Intermittent samples were taken, centrifuged at 6000 rpm for 7 minutes (EBA20, Hettich, Germany), and the supernatant was diluted with mobile phase A (described above) before being injected into the HPLC system for quantification. Results after 3 hours showed that 3.6% (for batch 3) and 2.2% (for batch 4) of liraglutide had been released, indicating minimal defects in the coating; therefore, any burst release is expected to be low.

[0321] In vitro studies To determine in vitro release, the material was suspended in 50 mL of 20 mM PIPES pH 7.2 + 1% Tween at a concentration of 0.5 mg liraglutide per mL of solvent in blue-capped flasks and incubated at 37°C for up to 72 hours. Intermittent samples were taken, filtered through a 0.2 μm PES filter, and then analyzed by HPLC using the same method as described for drug loading determination. The results for batch 3 (circles) and batch 4 (squares) are shown in Figure 5.

[0322] Example 6 Formulation III of the Invention Two suspensions of liraglutide-coated microparticles (Batch 3 and Batch 4, prepared according to the process described in Example 5 above) were prepared by adding an appropriate volume of Hyonate vet. (a 1 wt% hyaluronic acid solution in 1 mM phosphate buffer, pH 7.4) to test vials to achieve a liraglutide concentration equivalent to 10 mg / mL. The vials were then tapped at least 10 times to remove any material that may have settled to the bottom of the test vial. The formulations were then vortexed for approximately 60 seconds to ensure a uniform suspension.

[0323] Example 7 In vivo studies Nine male Goettingen SPF minipigs weighing approximately 10 kg on the day of treatment were supplied by Ellegaard Goettingen Minipigs A / S (Denmark). The animals were randomly divided into three animals per group.

[0324] The suspension (prepared as described in Example 6 above) or a comparator liraglutide solution (Victoza, Novo Nordisk, Denmark) diluted in normal saline was administered as a single dose subcutaneously through a 23G needle into the left side of the neck of each of three Goettingen minipigs per treatment group. Minipigs receiving Batch 3, Batch 4, or the comparator liraglutide solution correspond to Groups 5, 6, and 7, respectively, as identified in Table 3 below. [Table 3]

[0325] Blood samples for pharmacokinetics were collected from all animals on Day 1 and on subsequent days. Blood sampling was performed for all groups at 2, 4, 6, 12, 24, 48, 72, 96, and 144 hours post-treatment and pre-treatment. Additionally, blood was collected from animals in Groups 2 and 3 at 216, 360, 456, 552, and 648 hours post-treatment, corresponding to Days 10, 16, 20, 24, and 28.

[0326] Approximately 3 mL of blood samples were collected from the jugular / bijugular vein. Blood was sampled into vacutainers containing K2EDTA as an anticoagulant. The vacutainers were placed in ice water until centrifugation (10 min, 1270 g, +4°C). Each plasma sample was divided into two aliquots of approximately 0.5 mL, transferred to sponsor-provided cryotubes, and frozen at or below -70°C or on dry ice within 90 minutes of collection. The first set of samples was shipped on dry ice (approximately -70°C) using a thermologger to a Contributing Scientist for analysis. The second set of samples was stored at or below -70°C as backup samples. The backup samples were shipped within one month of receiving the last set of primary samples.

[0327] Liraglutide plasma concentrations were determined by LC-MS / MS. Study samples were prepared by pipetting 35 μL of minipig plasma into a 384-well plate and adding 35 μL of internal standard working solution using a TECAN liquid handling robot. The 384-well plate was then shaken for 1 minute. Protein precipitation solution was added, and the sample was shaken for 5 minutes and centrifuged. All samples were injected onto a UPLC-MS / MS system. Separation was performed on an ACQUITY Phenyl-Hexyl column, 1.7 μm, 2.1 x 50 mm, at 60°C using 0.3% formic acid as mobile phase A (MP A) and acetonitrile as mobile phase B (MP B) in water.

[0328] PK in Goettingen minipigs was assessed using non-compartmental analysis (NCA) utilizing the software Phoenix WinNonlin, version 8.3 (Certara, USA).

[0329] Nominal plasma sampling time points and nominal doses were used for non-compartmental PK analysis. Animal PK was calculated with the extravascular dose option in WinNonlin. 最大 Plasma concentrations below the LLOQ occurring before were treated as zero.

[0330] C最大 and t 最大 was derived from the observed plasma concentration data. AUC was estimated by integration of the plasma concentration versus time curve using linear interpolation for increasing plasma levels and logarithmic interpolation for decreasing plasma levels (linear up-log down method). AUC 最後 The measurable plasma concentration (AUC 最後 The area up to the final point indicating the sigma was calculated.

[0331] result The mean plasma concentrations of liraglutide over a 4-week period following a single subcutaneous administration of the various formulations (n=3) are shown in FIG.

[0332] Plasma pharmacokinetic parameters are also presented in Table 4 below as mean values ​​(with standard deviations provided in parentheses) per dose group, and C 最大 and t 最大 corresponds to the definition given above, "AUC 0-24h " is the AUC from time 0 to time 24 hours "AUC last ' is the AUC from time 0 to the time of the last detectable plasma concentration [Table 4]

[0333] Subcutaneous administration of two formulations of coated liraglutide microparticles (Groups 5 and 6) showed an initial absorption phase of liraglutide, followed by a slower and more prolonged absorption phase over at least 28 days compared to Group 7. The observed mean C for liraglutide 最大 was significantly lower in Groups 5 and 6 (coated liraglutide microparticle formulation, dose 1.0 mg) than that observed after subcutaneous administration of liraglutide solution (Group 7, dose 0.2 mg).

[0334] Plasma exposure to liraglutide appeared to be higher in Group 5 (Batch 3) than in Group 6 (Batch 4), which may be due to the thicker coating formulation in Group 6 compared to Group 5.

[0335] In summary, groups 5 and 6 have higher C 最大 The liraglutide solution showed a significantly different long-term release profile from that of the liraglutide solution, followed by a rapid decline in plasma concentration after administration.

[0336] Example 8 Coated liraglutide microparticles Liraglutide particles A batch with larger sized spray-dried particles of liraglutide was prepared using the process parameters defined below in a spray-drying apparatus similar to that used in Example 1. Otherwise, all other steps were performed according to the process described in Example 1.

[0337] For these formulations, liraglutide was dissolved in water for injection (WFI) to 5% (w / w) in a spray drying process. 5 g and 30 g batches were produced on a small-scale spray drying machine, PROCEPT SD3, using the equipment setting parameters as defined in Tables 5 (process parameter set points) and 6 (in-process result parameter ranges). [Table 5] [Table 6]

[0338] Batch 5 For batch 5 (297_5_LIR), larger-sized spray-dried liraglutide particles were used, as prepared by the process described above. The particle sizes of the larger-sized spray-dried liraglutide particles were determined as described above and were as follows: %D(10): 3.1 μm ± 4.3%, %D(50): 10.9 μm ± 4.0%, %D(90): 30.5 μm ± 6.8%.

[0339] Large spray-dried liraglutide particles from batch 5 were first coated with 10 layers of pure aluminum oxide by the following processes a to e: a. The valve on the piping between the pump and the ALD reactor was closed. b. Next, the valve on the trimethylaluminum (TMA) precursor bottle was opened for 1 second, filling the vaporized metal-containing precursor into the ALD reactor for 1 second. c. The valve to the precursor bottle was closed and the chamber was allowed to rest for 30 seconds (soak time) before opening the pump again to ensure the metal-containing precursor vapor reacted with the surface of the drug particles. d. The ALD reactor was then pumped for 9 seconds. e. The above steps a to d were repeated 20 times.

[0340] The chamber was then purged with a continuous flow of nitrogen to remove unreacted reagents and organic gases. Steps (a) through (e) above were then essentially repeated, except that water was used as the second reagent to form a distinct aluminum oxide layer on the surface of the active ingredient microparticles. This was followed by another purge pulse using a continuous flow of nitrogen to remove gaseous water and organic gases.

[0341] The above procedure was repeated nine times to form a total of 10 initial aluminum oxide atomic layers.

[0342] Following this, in step (b) above, 20 layers of zinc oxide were applied by repeating the above ALD steps using diethylzinc (DEZ) and then water as precursors to form a total of 20 zinc oxide layers, which provided a multilayer structure of pure aluminum oxide and zinc oxide (Al2O3:ZnO) with a total of 30 atomic layers, including 10 initial aluminum oxide atomic layers.

[0343] The powder was then removed from the ALD reactor and deagglomerated using a sonic sifter (Tsutsui Sonic Agitated Sifting Machine SW-20AT) with a sieve of 45 μm mesh size.

[0344] The resulting deagglomerated powder was reloaded into the ALD reactor, and the same procedure was followed to coat the particles with 10 layers of pure aluminum oxide, followed by 20 layers of pure zinc oxide, for a total of 30 atomic layers. After the final deagglomeration step, the same procedure was followed to coat the particles with 7 layers of pure aluminum oxide, followed by 20 layers of pure zinc oxide, and finally, 3 layers of pure aluminum oxide, for a total of 30 atomic layers of mixed oxide (Al2O3:ZnO:Al2O3). The deagglomeration and coating steps were repeated four times to prepare samples with five distinct coatings of pure aluminum oxide and zinc oxide.

[0345] Thus, batch 5 contained 5 x 30 layers (10 layers of Al2O3 and 20 layers of ZnO) and 1 x 30 layers (7 layers of Al2O3, 20 layers of ZnO and 3 layers of Al2O3).

[0346] Batch 6 For batch 6, spray dried liraglutide particles, as described in Example 1, were used.

[0347] The spray-dried liraglutide particles of batch 6 were first coated with 10 layers of pure aluminum oxide by the following processes a to e: a. The valve on the piping between the pump and the ALD reactor was closed. b. Next, the valve on the trimethylaluminum (TMA) precursor bottle was opened for 1 second, filling the vaporized metal-containing precursor into the ALD reactor for 1 second. c. The valve to the precursor bottle was closed and the chamber was allowed to rest for 30 seconds (soak time) before opening the pump again to ensure the metal-containing precursor vapor reacted with the surface of the drug particles. d. The ALD reactor was then pumped for 9 seconds. e. The above steps a to d were repeated 20 times.

[0348] The chamber was then purged with a continuous flow of nitrogen to remove unreacted reagents and organic gases. Steps (a) through (e) above were then essentially repeated, except that water was used as the second reagent to form a distinct aluminum oxide layer on the surface of the active ingredient microparticles. This was followed by another purge pulse using a continuous flow of nitrogen to remove gaseous water and organic gases.

[0349] The above procedure was repeated nine times to form a total of 10 initial aluminum oxide atomic layers.

[0350] This was followed by applying three layers of zinc oxide by repeating the ALD steps above using diethylzinc (DEZ) and then water as precursors in step (b) above, forming a total of three zinc oxide layers. This was followed by coating with one layer of aluminum oxide using the same precursors as described above in (b). This provided a mixed oxide layer with an atomic ratio of 1:3 (Al2O3:ZnO) for a total of four atomic layers.

[0351] This process was repeated a total of nine times to form a mixed oxide (1:3 Al2O3:ZnO) layer with a total of 50 atomic layers (30 layers of ZnO and 20 layers of Al2O3), including 10 initial aluminum oxide atomic layers.

[0352] The powder was then removed from the ALD reactor and deagglomerated using a sonic sifter (Tsutsui Sonic Agitated Sifting Machine SW-20AT) with a sieve of 20 μm mesh size.

[0353] The resulting deagglomerated powder was reloaded into the ALD reactor, and the same procedure was followed to coat the particles with 10 layers of pure aluminum oxide, followed by a layer of mixed oxide (1:3 Al2O3:ZnO), for a total of 50 atomic layers. After the final deagglomeration step, the same procedure was followed to coat the particles with 10 layers of pure aluminum oxide, followed by a layer of mixed oxide (1:3 Al2O3:ZnO), for a total of 50 atomic layers. The deagglomeration and coating steps were repeated three times to produce a sample with a base layer of pure aluminum oxide and four separate coatings of mixed oxide with a 1:3 Al2O3:ZnO atomic ratio.

[0354] Thus, batch 6 contained 4 x 50 layers, i.e. 4 x (10 layers of Al2O3 and 40 layers of 1:3 Al2O3:ZnO mixed oxide).

[0355] Batch 7 For Batch 7, larger size spray-dried liraglutide particles were used, as prepared by the process described above. The particle sizes of the larger size spray-dried material were determined as described above and were as follows: %D(10): 3.0 μm ± 1.2%, %D(50): 9.7 μm ± 2.4%, %D(90): 27.6 μm ± 6.1%.

[0356] Large spray-dried liraglutide particles from batch 7 were first coated with three layers of pure aluminum oxide by the following processes a to e: a. The valve on the piping between the pump and the ALD reactor was closed. b. Next, the valve on the trimethylaluminum (TMA) precursor bottle was opened for 1 second, filling the vaporized metal-containing precursor into the ALD reactor for 1 second. c. The valve to the precursor bottle was closed and the chamber was allowed to rest for 30 seconds (soak time) before opening the pump again to ensure the metal-containing precursor vapor reacted with the surface of the drug particles. d. The ALD reactor was then pumped for 9 seconds. e. The above steps a to d were repeated 20 times.

[0357] The chamber was then purged with a continuous flow of nitrogen to remove unreacted reagents and organic gases. Steps (a) through (e) above were then essentially repeated, except that water was used as the second reagent to form a distinct aluminum oxide layer on the surface of the active ingredient microparticles. This was followed by another purge pulse using a continuous flow of nitrogen to remove gaseous water and organic gases.

[0358] The above procedure was repeated twice to form a total of three initial aluminum oxide atomic layers.

[0359] This was followed by applying three layers of zinc oxide by repeating the ALD steps above using diethylzinc (DEZ) and then water as precursors in step (b) above, forming a total of three zinc oxide layers. This was followed by coating with one layer of aluminum oxide using the same precursors as described above in (b). This provided a mixed oxide layer with an atomic ratio of 1:3 (Al2O3:ZnO) for a total of four atomic layers.

[0360] This process was repeated five times to form a mixed oxide (1:3 Al2O3:ZnO) layer with a total of 27 atomic layers (18 layers of ZnO and 9 layers of Al2O3), including three initial aluminum oxide atomic layers.

[0361] The powder was then removed from the ALD reactor and deagglomerated using a sonic sifter (Tsutsui Sonic Agitated Sifting Machine SW-20AT) with a sieve of 32 μm mesh size.

[0362] The resulting deagglomerated powder was reloaded into the ALD reactor, and the same procedure was followed to coat the particles with three layers of pure aluminum oxide, followed by a layer of mixed oxide (1:3 Al2O3:ZnO), for a total of 27 atomic layers. After the final deagglomeration step, the same procedure was followed to coat the particles with three layers of pure aluminum oxide, followed by a layer of mixed oxide (1:3 Al2O3:ZnO), for a total of 27 atomic layers. The deagglomeration and coating steps were repeated three times to produce a sample with a base layer of pure aluminum oxide and four separate coatings of mixed oxide with a 1:3 Al2O3:ZnO atomic ratio.

[0363] Thus, batch 7 contained 4 x 27 layers, i.e. 4 x (3 layers of Al2O3 and 24 layers of 1:3 Al2O3:ZnO mixed oxide).

[0364] Batch 8 For batch 8, larger size spray-dried liraglutide particles were used, as prepared by the process described above. The particle sizes of the larger size spray-dried material were determined as described above and were as follows: %D(10): 3.1 μm ± 4.3%, %D(50): 10.9 μm ± 4.0%, %D(90): 30.5 μm ± 6.8%.

[0365] The larger size spray dried liraglutide particles of batch 8 were first coated with three layers of pure aluminum oxide by the following processes a to e: a. The valve on the piping between the pump and the ALD reactor was closed. b. Next, the valve on the trimethylaluminum (TMA) precursor bottle was opened for 1 second, filling the vaporized metal-containing precursor into the ALD reactor for 1 second. c. The valve to the precursor bottle was closed and the chamber was allowed to rest for 30 seconds (soak time) before opening the pump again to ensure the metal-containing precursor vapor reacted with the surface of the drug particles. d. The ALD reactor was then pumped for 9 seconds. e. The above steps a to d were repeated 20 times.

[0366] The chamber was then purged with a continuous flow of nitrogen to remove unreacted reagents and organic gases. Steps (a) through (e) above were then essentially repeated, except that water was used as the second reagent to form a distinct aluminum oxide layer on the surface of the active ingredient microparticles. This was followed by another purge pulse using a continuous flow of nitrogen to remove gaseous water and organic gases.

[0367] The above procedure was repeated twice to form a total of three initial aluminum oxide atomic layers.

[0368] This was followed by applying three layers of zinc oxide by repeating the ALD steps above using diethylzinc (DEZ) and then water as precursors in step (b) above, forming a total of three zinc oxide layers. This was followed by coating with one layer of aluminum oxide using the same precursors as described above in (b). This provided a mixed oxide layer with an atomic ratio of 1:3 (Al2O3:ZnO) for a total of four atomic layers.

[0369] This process was repeated five times to form a mixed oxide (1:3 Al2O3:ZnO) layer with a total of 27 atomic layers (18 layers of ZnO and 9 layers of Al2O3), including three initial aluminum oxide atomic layers.

[0370] The powder was then removed from the ALD reactor and deagglomerated using a sonic sifter (Tsutsui Sonic Agitated Sifting Machine SW-20AT) with a sieve of 45 μm mesh size.

[0371] The resulting deagglomerated powder was reloaded into the ALD reactor and the same procedure was followed to coat the particles with three layers of pure aluminum oxide, followed by a layer of mixed oxide (1:3 Al2O3:ZnO), for a total of 27 atomic layers. After the final deagglomeration step, the same procedure was followed to coat the particles with three layers of pure aluminum oxide, followed by a layer of mixed oxide (1:3 Al2O3:ZnO), for a total of 27 atomic layers. The deagglomeration and coating steps were repeated three times to produce samples with a base layer of pure aluminum oxide and four separate coatings of mixed oxide with a 1:3 Al2O3:ZnO atomic ratio.

[0372] Thus, batch 8 contained 4 x 27 layers, i.e. 4 x (3 layers of Al2O3 and 24 layers of 1:3 Al2O3:ZnO mixed oxide).

[0373] Batch characterization The particle size of the batches was determined by suspending the coated particles in a solution of 1% Span 85 (Sigma-Aldrich, MO, USA) in heptane (Merck, Germany). The particle size distribution measured by laser diffraction was Batch 5: %D(10): 7.2 μm ± 3.9%, %D(50): 17.9 μm ± 1.6%, %D(90): 36.2 μm ± 2.1%, Batch 6: Undecided, Batch 7: %D(10): 3.8 μm ± 1.3%, %D(50): 11.2 μm ± 1.9%, %D(90): 26.3 μm ± 6.8%, and Batch 8: %D(10): 4.5 μm ± 5.3%, %D(50): 13.0 μm ± 0.8%, %D(90): 30.0 μm ± 0.5%.

[0374] Drug loading for batches 5, 7 and 8 was determined by etching the samples in a solution of 88% water (PanReac, Spain) and 12% (v / v) phosphoric acid (Merck, Germany) to dissolve the coating, followed by dilution to 75 μg / mL in 75% (Rathburn, UK) + 0.1% trifluoroacetic acid (Merck, Germany) in water and loading onto a column Kinetex F5, 150x4.6mm, 2.6 μm particle size (Phenomenex Ltd., CA, USA), mobile phase A: 10% acetonitrile in water with 0.1% TFA, mobile phase B: 80% acetonitrile in water with 0.1% trifluoroacetic acid, injection volume 2 μL, autosampler temperature 20°C, oven temperature 25°C, flow rate 1.0 mL / min gradient elution, UV absorbance detection at 220 nm. Single-point calibration was used for the assay. The reference material by assay was 99.4% total liraglutide, with a purity of 99.1% by HPLC (Bachem AG, Switzerland), as determined by injection into an HPLC system for quantification (Prominence-I HPLC-UV-DAD, Shimadzu, Japan).

[0375] The drug loading for batch 6 was determined by etching the sample in a solution of 43% acetonitrile (Spectrascan, Scotland), 45% water (PanReac, Spain), and 12% (v / v) phosphoric acid (Merck, Germany) to dissolve the coating, followed by dilution to 160 μg / mL with 10% acetonitrile + 0.1% trifluoroacetic acid in water (Merck, Germany) (mobile phase A) on a column Kinetex C18 PS, 150 × 4.6 mm, 2.6 μm particle size (Phenomenex Ltd., CA, USA), mobile phase B: 80% acetonitrile in water + 0.1% trifluoroacetic acid, injection volume 10 μL, autosampler temperature 15°C, oven temperature 25°C, flow rate 1.0 mL / min gradient elution, UV absorbance detection at 220 nm. Single-point calibration was used for the assay. The retention time of liraglutide was 6.8 min. The reference material by assay was 99.4% total liraglutide, with a purity of 99.1% by HPLC (Bachem AG, Switzerland), as determined by injection into a quantification HPLC system (Prominence-I HPLC-UV-DAD, Shimadzu, Japan).

[0376] The drug loadings measured by HPLC-UV were 79.1 ± 0.5% for batch 5 (297_5_LIR), 62.8 ± 0.7% for batch 6 (278_4_LIR), 80.9 ± 0.2% for batch 7 (293_4_LIR), and 80.2 ± 0.3% for batch 8 (295_4_LIR).

[0377] To determine coating integrity, materials were suspended in dimethyl sulfoxide (Rathburn, UK) at a concentration of 0.4 mg liraglutide per mL of solvent and rotated on an overhead stirrer for 3 hours. Intermittent samples were taken and centrifuged at 6000 rpm for 7 minutes (EBA20, Hettich, Germany). The supernatant was diluted with mobile phase A (described above) and then injected into the HPLC system for quantification. Results after 3 hours showed 4.0% liraglutide released for batch 5, 4.6% for batch 6, 5.2% for batch 7, and 7.8% for batch 8, indicating minimal defects in the coating; therefore, any burst release is expected to be low.

[0378] In vitro studies To determine in vitro release, materials were suspended in 50 mL of 20 mM PIPES pH 7.2 + 1% Tween at a concentration of 0.5 mg liraglutide per mL of solvent in blue-capped flasks and incubated at 37°C for up to 72 hours. Intermittent samples were taken, filtered through a 0.2 μm PES filter, and then analyzed by HPLC using the same method described for drug loading determination. Results for batch 5 (circles), batch 6 (squares), batch 7 (diamonds), and batch 8 (triangles) are shown in Figure 7.

[0379] Example 9 Formulation IV of the Invention Four suspensions of liraglutide-coated microparticles (Batch 5, Batch 6, Batch 7, and Batch 8, prepared according to the process described in Example 8 above) were prepared by adding the appropriate volume of Hyonate vet. (a 1 wt% hyaluronic acid solution in 1 mM phosphate buffer, pH 7.4) to test article vials to achieve a liraglutide concentration equivalent to 10 mg / mL. The vials were then tapped at least 10 times to remove any material that may have settled to the bottom of the test vial. The formulations were then vortexed for approximately 60 seconds to ensure a uniform suspension.

[0380] Example 10 In vivo studies Sixteen male Sprague-Dawley rats were divided into four groups of four animals per group according to Table 7 below. The animals were weighed and administered a single subcutaneous injection of the test article using a 23G needle. Blood samples for plasma isolation were collected at 3, 6, 12, 24, 36, 48, 72, 120, 168, 251, 384, 480, 576, and 672 hours after administration. Visual inspection of the administration site was performed within 1 hour of dosing and at blood sampling times from 24 hours onward. After the final blood sample was collected, the animals were euthanized. [Table 7]

[0381] Blood samples (approximately 0.2 mL) were collected from the jugular vein into KEDTA (dipotassium ethylenediaminetetraacetic acid) tubes at 1, 3, 6, 12, 24, 48, 72, 120, 168, 251, 384, 480, 576, and 672 hours after dosing for Groups 1-3, and at 1, 2, 3, 6, 9, 12, 24, and 48 hours after subcutaneous dosing for Group 4. Actual sampling times were recorded. Plasma was separated by centrifugation (1500 g, 10 minutes at 4°C) as soon as practical after blood sampling and stored at -80°C until analysis.

[0382] After study completion, all plasma samples were shipped for analysis deep frozen on dry ice. Animals were sacrificed on the final day of the study.

[0383] The plasma concentration of liraglutide was determined by LC-MS / MS. Study samples were prepared by pipetting 35 μL of rat plasma into a 96-well plate and adding 75 μL of internal standard working solution using an EVO-2 liquid handling robot (Tecan, Austria). The 96-well plate was then shaken for 15 minutes and centrifuged. All samples were then injected onto a UPLC-MS / MS system (Waters, MA, USA). Separation was performed using an ACQUITY UPLC Protein BEH C4 column, 300 Å, 1.7 μm, 2.1 mm × 50 mm (Waters, MA, USA) at 60 °C using 0.3% formic acid in water as mobile phase A and acetonitrile as mobile phase B.

[0384] The pharmacokinetic (PK) analysis of liraglutide in plasma was evaluated using non-compartmental analysis (NCA) utilizing the software Phoenix WinNonlin, version 8.3 (Certara, USA). 最大 and t 最大 was derived from the observed plasma concentration data. AUC was estimated by integration of the plasma concentration versus time curve using linear interpolation for increasing plasma levels and logarithmic interpolation for decreasing plasma levels (linear up-log down method). AUC ∞ The measurable plasma concentration (AUC 最後 ) and then calculate the area up to the last point that shows the concentration in the last quantifiable sample and lambda z and extrapolate to infinity using the curve t 1 / 2,z The first-order rate constant associated with the terminal portion of z was calculated by

[0385] result Dose-normalized plasma concentrations of liraglutide over 1 week (FIG. 8) and 42 days (FIG. 9) following a single subcutaneous administration of the various formulations are shown in FIGS. 8 and 9.

[0386] The plasma pharmacokinetic parameters are also presented in Table 8 below as the mean values ​​of the four groups (standard deviations are provided in parentheses), with the parameter definitions corresponding to those above: · “Fr.Rel. 0-72h " is the fraction released during the first 72 hours of the area under the concentration versus time curve to infinity expressed as a percentage. [Table 8]

[0387] As can be seen in Figure 8, subcutaneous administration of the four formulations showed an initial absorption phase of liraglutide, followed by a slower, prolonged absorption phase over at least 7 days. In particular, Figures 8 and 9 show that the formulation administered to Group 11 showed a higher release in the first 7 days, followed by a lower release over 42 days, compared to Groups 8-10, which showed sustained release over 42 days. Group 8 (Batch 5, i.e., corresponding to the Al+Zn coating) showed a lower C compared to Group 11 (Batch 8, i.e., corresponding to the 1:3 AZO coating). 最大 Both of these formulations were manufactured using 10.9 μm (%D50) liraglutide particles as raw material.

Claims

1. 1. A pharmaceutical formulation useful for treating a metabolic disorder or condition, comprising: a plurality of particles suspended in a carrier system, said particles comprising: (a) have an average diameter by weight, number, or volume of about 10 nm to about 700 μm; (b)(i) zinc oxide; (ii) a solid core comprising at least one glucagon-like peptide-1 receptor agonist or a pharmaceutically acceptable salt thereof, at least partially coated with a coating of inorganic material comprising a mixture of: The atomic ratio ((i):(ii)) is at least about 1:10 and not more than about 10:

1.

2. 2. The formulation of claim 1, wherein the atomic ratio ((i):(ii)) is at least about 1:1 and not more than about 6:

1.

3. 3. The formulation of claim 1 or claim 2, wherein the coated particle comprises one or more discrete layers surrounding the core, at least one of the discrete layers comprising a mixture of zinc oxide and one or more other metal and / or semi-metal oxides.

4. 10. A formulation according to any one of the preceding claims, wherein the coated particle comprises two or more distinct layers surrounding the core.

5. 5. The formulation of claim 4, wherein at least one of the individual layers consists essentially of a single metal and / or semi-metal oxide.

6. 6. The formulation of claim 5, wherein at least one of the individual layers of a single metal and / or semi-metal oxide is applied directly to the core before and / or after application of the one or more layers comprising the mixture of zinc oxide and one or more other metal and / or semi-metal oxides.

7. A formulation according to any one of claims 4 to 6, wherein the number of said individual layers is up to four layers of a single metal and / or semi-metal oxide.

8. A formulation according to any one of claims 4 to 7, wherein the single metal and / or semi-metal oxide is aluminium oxide.

9. 10. The formulation of any one of the preceding claims, wherein the core consists essentially of at least one glucagon-like peptide-1 receptor agonist, or a pharmaceutically acceptable salt thereof.

10. 10. A formulation according to any one of the preceding claims, wherein the mean diameter by weight, number or volume of the particles is in the amount of 1 μm to about 50 μm.

11. 10. A formulation according to any one of the preceding claims, wherein two or more individual layers of the mixture of oxides are applied sequentially to the core.

12. The formulation of claim 11, wherein 3 to 10 individual layers of the mixture of oxides are applied.

13. 10. The formulation according to any one of the preceding claims, wherein the total thickness of the mixed oxide coating is from about 0.5 nm to about 2 μm.

14. 14. The formulation of any one of claims 4 to 13, wherein the maximum thickness of each individual discrete layer of oxide coating is about 1 / 100 of the average diameter by weight, number or volume of the core, including any other individual layers previously applied to the core.

15. 10. A formulation according to any one of the preceding claims, wherein the ratio of zinc oxide to other metal and / or semi-metal oxides is from about 2:1 to about 5:

1.

16. 10. A formulation according to any one of the preceding claims, wherein the one or more other metal and / or semi-metal oxides are selected from aluminium oxide and / or silicon dioxide.

17. 10. A formulation according to any one of the preceding claims in the form of a sterile injectable dosage form.

18. 18. The formulation of claim 17 in a form that can be administered via a surgical administration device to form a depot formulation.

19. 10. The formulation of any one of the preceding claims, wherein the at least one glucagon-like peptide-1 receptor agonist is liraglutide.

20. 19. The formulation of any one of claims 1 to 18, wherein the at least one glucagon-like peptide-1 receptor agonist is semaglutide.

21. 10. A process for preparing a formulation according to any one of the preceding claims, comprising: A process for preparing a formulation, wherein the coated particles are produced by applying a layer(s) of a mixed oxide coating material to the core and / or a previously coated core by atomic layer deposition.

22. (i) coating the solid core with a first discrete layer of a mixed oxide coating material; (ii) then subjecting said coated cores from step (i) to a deagglomeration process step; (iii) then coating the deagglomerated coated cores from step (ii) with a second, discrete layer of a mixed oxide coating material; 22. The process of claim 21, wherein (iv) steps (ii) and (iii) are repeated to obtain as many individual layers as required.

23. 23. The process of claim 22, wherein the deagglomeration step performed during application of the coating comprises sieving.

24. 24. The process of claim 23, wherein the sieving comprises vibratory sieving.

25. 25. The process of claim 24, wherein the vibratory screening includes controlling a vibrating probe coupled to a screen.

26. 24. The process of claim 23, wherein the sieving comprises sonic sieving.

27. A process for preparing a formulation according to any one of claims 1 to 20, wherein the coated particles are mixed with the carrier system after coating.

28. 21. An injectable dosage form comprising a formulation according to any one of claims 1 to 20 connected to an injection means and / or contained in a reservoir associated with the injection means.

29. 29. The dosage form of claim 28, which is a surgical administration device forming a depot formulation.

30. 30. The dosage form of claim 28 or claim 29, which is an auto-injector.

31. A formulation according to any one of claims 1 to 20 or a dosage form according to any one of claims 28 to 30 for use in the treatment of a metabolic disorder or condition.

32. Use of a formulation according to any one of claims 1 to 20 or a dosage form according to any one of claims 28 to 30 for the manufacture of a medicament for the treatment of a metabolic disorder or condition.

33. 1. A method for treating a metabolic disorder or condition, comprising: A method comprising administering a formulation according to any one of claims 1 to 20 or a dosage form according to any one of claims 28 to 30 to a patient in need of such treatment.

34. 34. The formulation for use according to claim 31, the use according to claim 32, or the method according to claim 33, wherein the metabolic disorder or condition is selected from type 2 diabetes, obesity, and / or co-morbidities.

35. 35. The formulation, use or method for use according to any one of claims 31 to 34, wherein after injection the formulation provides a depot formulation in which the at least one glucagon-like peptide-1 receptor agonist is released over a period of from 3 months to about 1 year.

36. 36. The formulation, use or method for use of claim 35, wherein the total exposure of the at least one glucagon-like peptide-1 receptor agonist is at least about 80% to about 125% of the total exposure obtained from a dosing regimen comprising administering the at least one glucagon-like peptide-1 receptor agonist by daily or weekly injection over the same period of time.

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