Novel pharmaceutical composition comprising glucagon-like peptide-1 receptor agonist

Abstract

There is provided a pharmaceutical formulation useful for the treatment of a metabolic disorder or condition, the pharmaceutical formulation comprising a plurality of particles suspended in a carrier system, the particles: (a) having an average diameter based on weight, quantity or volume of between about 10 nm and about 700 [mu] m; and (b) comprises a solid core comprising at least one glucagon-like peptide-1 receptor agonist or a pharmaceutically acceptable salt thereof, said solid core being at least partially coated by an inorganic material coating comprising a mixture of (i) zinc oxide; and (ii) one or more other metal oxides and / or metalloid oxides, wherein the atomic ratio ((i): (ii)) is at least about 1: 10 and up to and including about 10: 1. The mixed oxide coated particles are preferably synthesized by vapor phase coating techniques, such as atomic layer deposition. The formulations may provide delayed or sustained release of a glucagon-like peptide-1 receptor agonist to treat metabolic disorders or conditions, such as type 2 diabetes and / or obesity, without burst release effects. The glucagon-like peptide-1 receptor agonist is preferably a liraglutide (liraglutide), and the glucagon-like peptide-1 receptor agonist is preferably a liraglutide (liraglutide).

Description

TECHNICAL FIELD

[0001] The present invention relates to a novel formulation for use in, for example, the field of drug delivery, and in particular for 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 acknowledgement that the document is part of the state of the art or is common general knowledge.

[0003] Diabetes mellitus is a metabolic disorder characterized by chronic hyperglycemia with disturbances of carbohydrate, fat, and / or protein metabolism due to a defect in insulin secretion, insulin action, or both. Diabetes mellitus is one of the oldest diseases described, found in Egyptian manuscripts dating from about 1500 BC. However, the importance of insulin for this disease was not established until the 1920s.

[0004] The most prevalent form of diabetes (approximately 90% of all patients) is type 2 diabetes, which has become a global epidemic in recent years, causing a significant healthcare burden. It is projected that the number of patients worldwide diagnosed with this condition will exceed 500 million by 2035.

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

[0006] Type 2 diabetes usually begins in late life (e.g., middle age or old age), but the incidence is known to be increasing in younger people. It not only shortens life expectancy by itself, but also often 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 largely believed to be related to diet, especially a diet that includes excess intake and / or insufficient energy expenditure from exercise. In this regard, type 2 diabetes and obesity are interdependent, with weight loss improving the prognosis of overweight type 2 diabetic patients and obese individuals. In fact, the global burden of type 2 diabetes is believed to be mainly due to the increase in obesity, which itself has become a global epidemic in recent decades.

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

[0009] Like the glucose-dependent insulinotropic peptide (GIP), GLP-1 is a gut incretin, i.e., it lowers blood glucose levels in a glucose-dependent manner by enhancing the secretion of insulin. GLP-1 based therapies are associated with weight loss and reduced risk of hypoglycemia, and thus are of particular importance for the treatment of patients with type 2 diabetes.

[0010] Endogenous GLP-1 is rapidly degraded, with a half-life of only about 2 minutes. Accordingly, a great deal of research has been conducted into the development of GLP-1 receptor agonists to increase the activity of GLP-1. In this regard, GLP-1 and analogs and fragments thereof (hereinafter collectively referred to as ‘GLP-1 receptor agonists’ or ‘GLP1RAs’) are useful as insulinotropic agents, and thus for the treatment of diabetes (including type 1 diabetes, and especially type 2 diabetes) and / or obesity.

[0011] Approved GLP1RAs include exenatide (Byetta®) (Amylin Pharmaceuticals, Inc. / Eli Lilly and Company), liraglutide (Victoza®) (Novo Nordisk A / S), lixisenatide (Amlynan®) (Sanofi), albiglutide (Soliqua®) (Takeda), dulaglutide (Trulicity®) (Eli Lilly and Company), semaglutide (Ozempic®) (Novo Nordisk A / S), taspoglutide (Zafatek®) (Servier), and exenatide extended-release (Bydureon®) (Amylin Pharmaceuticals, Inc. / Eli Lilly and Company).

[0012] The above GLP1RA-containing products are administered by subcutaneous injection into adipose tissue, typically into the abdomen or upper thighs. Existing injectable formulations comprising GLP1RAs are typically injected once or twice daily (classified as short-acting GLP1Ras) or weekly (long-acting GLP1Ras), which is still relatively frequent.

[0013] According to a review article by Sikirica et al., Diabetes Metab. Syndr. Obes., 10, 403 (2017), the percentage of patients reporting problems with injecting GLP1RAs is higher than average physician estimates. Common patient-reported problems include injection pain and / or inconvenience of regular injections. The most common complaints reported by patients (56%) and physicians (32.6%) were that oral medications are superior to injections. ​​​​​​​

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

[0015] While semaglutide has been formulated as an oral drug in the form of (NovoNordisk), the compound exhibits extremely low oral bioavailability and must be taken on an empty stomach with no more than 120 mL of water daily and it is recommended to be followed by eating within 30 and 60 minutes. All of this can also lead to patient compliance issues, as well as an increased risk of adverse events.

[0016] In chronic conditions like type 2 diabetes or obesity, it would be advantageous to provide a sustained release composition in which the active ingredient is released in the body at a desired and predictable rate over weeks or months after injection, to ensure better patient compliance and possibly a better pharmacokinetic profile. An increase in patient compliance over time is expected to help improve overall treatment effectiveness (and thus make patients generally healthier) and save costs for healthcare and society.

[0017] In any case, a more convenient, lower injection frequency GLP1RA formulation would be expected to be an extremely attractive treatment option for all type 2 diabetes patients who are currently being treated with GLP1RAs, even for those patients for whom adherence to their prescribed dosage regimen is not an issue.

[0018] In the case of any sustained release composition, it is crucial that its release profile shows minimal initial rapid release of the active ingredient, i.e. high drug concentration in the plasma shortly after administration. Such a ‘sudden’ release would lead to undesirably high concentrations of the active ingredient and can be dangerous in the case of drugs with a narrow therapeutic window or drugs that are toxic at high plasma concentrations.

[0019] In the case of an injectable suspension of the active ingredient, it is also important to control the size of the suspended particles so that they can pass through the needle for injection. If there are large aggregated particles, they will not only clog the needle through which the suspension is injected, but also will not form a stable suspension within the injected liquid (i.e. they tend to settle to the bottom of the injected liquid instead).

[0020] Thus, there is an unmet clinical need for more durable, more effective, and / or improved drug delivery systems comprising GLP1 RAs in the treatment of conditions like type 2 diabetes and obesity.

[0021] Atomic layer deposition (ALD) is a technique for depositing thin films comprising a variety of materials, including organic materials, biological materials, polymeric materials, and especially inorganic materials, such as metal oxides, on solid substrates. Atomic layer deposition is an enabling technology for atomic and near-atomic scale manufacturing (ACSM) of materials, structures, devices, and systems in a variety of applications (see, e.g., Zhang et al. Nanomanuf. Metrol. 2022, https: / / doi.org / 10.1007 / s41871-022-00136-8). Based on its self-limiting nature, ALD can achieve atomic-level thickness controlled only by adjusting the number of growth cycles. Furthermore, multiple layers can be deposited, and the properties of each layer can be tailored at the atomic level.

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

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

[0024] Solid coating protects the coated substrate from chemical reactions (decomposition) and physical changes. ALD can also potentially be used to control the release rate of the substrate material within a solvent, which makes it potentially useful in formulations of active pharmaceutical ingredients.

[0025] In ALD, a first precursor, which can be metal-containing, is fed into the ALD reactor chamber (in a so-called ‘precursor pulse’), and the first precursor forms an adsorbed atomic or molecular monolayer at the surface of the substrate. The excess first precursor is then purged from the reactor, and then a second precursor, such as water, is pulsed into the reactor. This reacts with the first precursor, resulting in the formation of a monolayer of, e.g., a metal oxide, on the substrate surface. A subsequent purge pulse is followed by another first precursor pulse, and thus a new cycle of the same events (a so-called “ALD cycle”) is started.

[0026] The thickness of the thin film coating is controlled, inter alia, by the number of ALD cycles performed.

[0027] In the normal ALD method, since only atomic or molecular monolayers are generated during any given cycle, no discernible physical interfaces are formed between these monolayers, which essentially become a continuum at the surface of the matrix.

[0028] International Patent Application No. WO 2014 / 187995 describes a method in which multiple ALD cycles are performed, followed by periodic removal of the resulting coated matrix from the reactor and a redispersion / stirring step to provide a new surface that can be used for precursor adsorption.

[0029] The stirring step is primarily to address the observed issues with nanoparticles and microparticles, specifically the aggregation of particles during ALD coating, leading to "pinholes" formed at the contact points between these particles. A redispersibility / stirring step is performed by placing the coated matrix in water and subjecting it to sonication, which results in deagglomeration and the breaking down of contact points between the individual particles of the coated active material.

[0030] The particles were then returned to the reactor, and the ALD coating step and powder deagglomeration step were repeated three times, for a total of four series of cycles. This method has been found to allow the formation of coated particles that are largely free of pinholes (see also Hellrup et al., International Journal of Pharmaceutical Sciences, 529, 116 (2017)).

[0031] A novel injectable composition comprising one or more GLP1RAs has now been prepared, wherein microparticles containing one or more GLP1RAs are coated with a specific mixed oxide coating layer using an ALD, wherein the coated particles are suspended in a carrier. This composition produces favorable pharmacokinetic profiles by releasing the active ingredient over a prolonged period of time to provide a therapeutically effective level of drug in systemic circulation without any significant initial burst release effect. Summary of the Invention

[0032] According to a first aspect of the present invention, a pharmaceutical formulation is provided for treating metabolic disorders or symptoms, the pharmaceutical formulation comprising a plurality of particles suspended in a carrier system, the particles being:

[0033] (a) Having an average diameter based on weight, number, or volume between about 10 nm and about 700 μm; and

[0034] (b) Containing a core, the core comprising one or more GLP1RAs or pharmaceutically acceptable salts thereof, the core being at least partially coated with an inorganic material coating comprising a mixture of the following:

[0035] (i) Zinc oxide (ZnO); and

[0036] (ii) one or more other metal oxides and / or metal-like oxides.

[0037] Where the atomic ratio ((i):(ii)) is at least about 1:10 and at most and includes about 10:1,

[0038] The formulation is referred to below as 'the formulation of the present invention'.

[0039] Preferably, the atomic ratio ((i):(ii)) is at least about 1:1 and at most and includes about 6:1.

[0040] Coatings comprising a mixture of zinc oxide and one or more other metal oxides and / or metal-like oxides are referred to below as 'mixed oxide' coatings or coating materials.

[0041] Those skilled in the art will readily understand that the term 'solid' includes any form of substance that retains its shape and density without restriction, and / or whose molecules are generally compressed as tightly as possible within the limits allowed by the repulsive forces between them. A solid core has at least one solid outer surface on which a coating material layer may be deposited. The interior of a solid core may also be solid or may be hollow. For example, if the particles are spray-dried before being placed in a reactor vessel, the particles may be hollow due to the spray-drying technique.

[0042] The core of the formulation of the present invention comprises one or more GLP1RAs or pharmaceutically acceptable salts thereof, and in this respect, the core may consist substantially of the one or more GLP1RAs or their salts, or may include the one or more GLP1RAs or their salts along with other excipients or other active ingredients.

[0043] "Constitutes substantially GLP1RA or a pharmaceutically acceptable salt thereof" means that the core is substantially composed only of one or more GLP1RAs or salts thereof, i.e., the core does not contain non-biologically active substances such as excipients, carriers, etc. (see below) and other active substances. This means that the core may contain less than about 5%, such as less than about 3%, including less than about 2%, such as less than about 1%, of such other excipients and / or active substances.

[0044] In an alternative, a core containing one or more GLP1RAs or pharmaceutically acceptable salts thereof may include an active ingredient mixed with one or more pharmaceutical ingredients, which may include pharmaceutically acceptable excipients such as adjuvants, diluents or carriers, and / or may include other bioactive ingredients.

[0045] The non-biologically active adjuvants, diluents, and carriers that can be used to coat the core according to the present invention may include pharmaceutically acceptable substances that are soluble in water, such as carbohydrates, for example 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.

[0046] Such excipients are preferably incorporated into a core containing GLP1RA or a pharmaceutically acceptable salt thereof by a 'spray drying' method. Those skilled in the art will understand that a 'spray drying' method includes any method of producing a dry powder from a liquid comprising a solution or suspension (including a slurry), which involves using hot gas to rapidly dry the liquid stream to convert it into an evaporating solvent and solid particles containing solutes previously dissolved in the solution and / or particles previously suspended in the evaporating liquid.

[0047] GLP1RA and its pharmaceutically acceptable salts may exist in crystalline, partially crystalline, and / or amorphous states. GLP1RA and its pharmaceutically acceptable salts may be solid or convertible to solid at approximately room temperature (e.g., approximately 18°C) and approximately atmospheric pressure, regardless of their physical form. The active agent (and optionally other pharmaceutical ingredients mentioned above) should also remain in solid form when coated in, for example, an ALD reactor, and should not physically or chemically degrade to a perceptible degree (i.e., not exceeding approximately 10% w / w) when coated with or covered by a mixed metal oxide coating material.

[0048] 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, followed by removal of the solvent or medium using standard techniques (e.g., under vacuum, by freeze-drying, or by filtration). Salts can also be prepared using techniques known to those skilled in the art, such as by exchanging an antiion of the active ingredient in salt form with another antiion (e.g., using a suitable ion exchange resin).

[0049] Specific salts that may be mentioned include, for example, 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, specifically acetic acid, more specifically trifluoroacetic acid, etc., and acid addition salts.

[0050] Preferred GLP1RAs include exenatide, lixilamide, abiglutide, dulaglutide, tesipatide, more preferably dexmemagglutide and especially liraglutide, all of which are known in the art and each of which can be synthesized by classical liquid-phase techniques, solid-phase methods or by recombinant peptide production.

[0051] The formulations of the present invention contain a pharmacologically effective amount of one or more GLP1RAs or a pharmaceutically acceptable salt thereof. Preferably, the core of the formulation of the present invention contains a pharmacologically effective amount of one or more (preferably one) GLP1RAs or a salt thereof.

[0052] The term "pharmacologically effective amount" refers to the amount of a relevant GLP1RA (alone or in combination) or its salt, whether administered alone or in combination with another active ingredient (which may or may not be another GLP1RA), that would confer the desired physiological change (such as a therapeutic effect) to the treated patient. Such a biological or medical response or such effect in a patient can be subjective (i.e., the subject gives an indication or feeling of the effect) and includes at least partial relief of symptoms of the disease or condition being treated, or cure or prevention of said disease or condition, or can be objective (i.e., measurable by some test or biomarker).

[0053] Therefore, the dose of GLP1RA / its salt that can be administered to a patient should be sufficient to influence a therapeutic response within a reasonable and / or relevant timeframe. Those skilled in the art will recognize that the selection of accurate dosage and composition, as well as the most suitable delivery method, will be influenced not only by the pharmacological properties of the active ingredient and the composition of the formulation, but also, in particular, by the route of administration, the nature and severity of the condition being treated, the recipient's physical condition and mental acuity, the patient's age, condition, weight, sex, and response, the stage / severity of the disease, and genetic differences between patients.

[0054] The dosage of GLP1RA / its salt can also be determined by the timing and frequency of administration. In any case, a licensed physician or other technician will be able to routinely determine the appropriate actual dosage of GLP1RA and / or its salt best suited for an individual patient.

[0055] GLP1RA is released over an extended period of time. The period of time may be at least about 3 days, such as about 5 days or about 7 days, and at most about one year, such as about 10 months, about 8 months, about 6 months, about 4 months, about 3 months or about 4 weeks, such as about 3 weeks (e.g., about 2 weeks).

[0056] Therefore, a suitable dose of GLP1RA or a pharmaceutically acceptable salt thereof in the formulation of the present invention can provide a plasma concentration-time profile that provides an exposure (AUC, defined as, for example, AUC) that provides at least the same therapeutic effect as that obtained from currently commercial subcutaneous injection of the relevant GLP1RA used in current clinical practice. last (The area under the curve of plasma concentration versus time up to the last detectable concentration over the extended time period) or more preferably, AUC ∞ (Up to infinity, the area under the plasma concentration-time curve)).

[0057] Regarding AUC ∞ In this regard, the formulations of the present invention may be able to provide no more than the total exposure (AUC) within any of the time periods mentioned above. ∞ The total exposure to GLP1RA in plasma is between approximately 80% and approximately 125%, obtained from relevant current care / dosing protocols for the administration of GLP1RA via injection (e.g., daily or weekly) via intravenous, intramuscular, or preferably subcutaneous injection.

[0058] More preferably, the total exposure to relevant GLP1RA (e.g., AUC) within any of the time periods mentioned above. ∞ This can be the total exposure (e.g., AUC) obtained from the current standard of care / dosing regimen for administration via injection of 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%.

[0059] This will enable the dosage of the relevant GLP1RA or its pharmaceutically acceptable salt within the formulation of the present invention to provide, or be able to provide, a daily or weekly dose (as applicable) ranging from about 10% (e.g., about 15%) to about 80% (e.g., about 70%, such as about 65%) of the daily or weekly dose administered within the current standard of care (i.e., the average dose released daily or weekly from the formulation during any of the time periods mentioned above after injection), where the current standard of care is daily or weekly injection therapy of the relevant GLP1RA compound or its salt (calculated relative to the free compound).

[0060] Therefore, the total dose of the formulation of the present invention, which contains liraglutide or a pharmaceutically acceptable salt thereof, injected into a patient can range from about 15 mg (e.g., about 270 mg) to about 1000 mg.

[0061] Therefore, the total dose of the formulation of the present invention, which contains zemagglutinin or a pharmaceutically acceptable salt thereof, injected into a patient can range from about 1 mg (e.g., about 30 mg) to about 100 mg.

[0062] Because the formulations of this invention provide a steady-state release of the relevant GLP1RA after injection, this means that the average C max (The maximum concentration observed in the plasma concentration-time curve) will be less than the C obtained from the current standard of care / dosing regimen via injection-related GLP1RA administration. max (For example, liraglutide is 20 ± 15 ng / mL, and sesmagglutide is 210 ± 30 ng / mL). For formulations of the present invention containing liraglutide, the average C max It can be between about 20 ng / mL and about 100 ng / mL. For formulations of the present invention containing cemaglutide, the average C1 max It can range from approximately 75 ng / mL to approximately 410 ng / mL.

[0063] The core of the formulation containing solid GLP1RA of the present invention is provided in the form of nanoparticles (or more preferably microparticles). Preferably, the average diameter based on weight, quantity, or volume is between about 50 nm (e.g., about 100 nm, such as about 250 nm) and about 30 μm, for example between about 500 nm and about 100 μm, more specifically about 1 μm (such as about 5 μm, including about 7 μm, about 9 μm, about 10 μm, or about 15 μm) to about 50 μm, such as about 25 μm, for example about 20 μm.

[0064] As used herein, those skilled in the art will understand that the term 'weight-based average diameter' includes an average particle size characterized and defined by a particle size distribution by weight, i.e., a distribution in which the present fraction (relative amount) in each size class is defined as a distribution of weight fractions obtained, for example, by sieving (e.g., wet sieving). As used herein, those skilled in the art will understand that the term 'quantity-based average diameter' includes an average particle size characterized and defined by a particle size distribution by quantity, i.e., a distribution in which the present fraction (relative amount) in each size class is defined as a distribution of quantity fractions measured, for example, by microscopy. As used herein, those skilled in the art will understand that the term 'volume-based average diameter' includes an average particle size characterized and defined by a particle size distribution by volume, i.e., a distribution in which the present fraction (relative amount) in each size class is defined as a distribution of volume fractions measured, for example, by laser diffraction. Those skilled in the art will also understand that other suitable ways of expressing average diameter exist, such as area-based average diameter, and these other expressions of average diameter are interchangeable with those used herein. Particle size can be measured using other instruments well known in the art, such as those sold by Malvern Instruments, Ltd., Worcestershire, UK, and Shimadzu Corporation, Kyoto, Japan.

[0065] The particles may be spherical, i.e., the aspect ratio of the particles is less than about 20, more preferably less than about 10, such as less than about 4, and especially less than about 2, and / or in at least about 90% of the particles, the variation in radius (measured from the center of gravity to the particle surface) may not exceed about 50% of the average value, such as not exceeding about 30% of the value, for example not exceeding about 20% of the value.

[0066] However, according to the invention, particles can also be coated in any shape. For example, particles with irregular shapes (e.g., 'raisin' shapes), needle-like, flake-like, or cuboid shapes can be coated. For non-spherical particles, the size can be expressed as, for example, the size of a corresponding spherical particle of the same weight, volume, or surface area. According to the invention, hollow particles and particles with pores, cracks, etc., such as fibrous particles or 'entangled' particles, can also be coated.

[0067] Particles can be obtained in a form suitable for coating, or by, for example, particle size reduction processes, such as wet milling, dry milling, air jet milling (including cryogenic micronization), ball milling, such as planetary ball milling, and using end roller mills, roller mills, vibratory mills, hammer mills, fluid energy mills, pin mills, etc. (e.g., crushing, cutting, grinding, or polishing) to a specific weight-based average diameter (as defined above). Alternatively, particles can be directly prepared into suitable sizes and shapes, for example by spray drying, freeze drying, spray freeze drying, vacuum drying, precipitation, including using supercritical fluids or other top-down methods (i.e., reducing the size of large particles by, for example, grinding), or bottom-up methods (i.e., increasing the size of small particles by, for example, sol-gel technology, crystallization, etc.). Alternatively, nanoparticles can be prepared using well-known techniques such as gas condensation, friction, chemical precipitation, ion implantation, pyrolysis, hydrothermal synthesis, etc.

[0068] It may be necessary (depending on how the particles containing the core are initially provided) to wash and / or clean the particles to remove impurities that may have originated from their production, and then the particles are dried. Drying can be performed using a variety of 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. If drying has been performed, the core can be deagglomerated by grinding, screening, milling, and / or dry ultrasonic treatment. Alternatively, the core can be treated to remove any volatile substances that may be absorbed onto its surface, for example, by exposing the particles to a vacuum and / or elevated temperatures.

[0069] The surface of the core can be chemically activated prior to the application of the first coating layer, for example by treatment with hydrogen peroxide, ozone, radical-containing reactants, or by plasma treatment, to generate oxygen radicals at the core surface. This, in turn, can create adsorption / nucleation sites on the core that are favorable for (e.g., ALD) precursors.

[0070] The core containing GLP1RA may be at least partially coated with a coating material comprising zinc oxide and one or more other metal oxides and / or metal-like oxides, wherein the atomic ratio of zinc oxide to the other oxide is at least about 1:10 (e.g., at least about 1:6 or about 1:4, such as at least about 1:2), preferably at least about 1:1 (e.g., at least about 1.5:1, such as at least about 2:1), including at least about 2.25:1, such as 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 at most (i.e. not exceeding) and including about 10:1, including at most about 8:1, about 7:1, about 6:1, about 5.5:1 or at most about 5:1, such as at most about 4.5:1, including at most about 4:1 (e.g., at most about 3.75:1).

[0071] Preferred methods for applying coating to a core containing a bioactive agent include vapor phase techniques such as ALD or related techniques such as atomic layer epitaxy (ALE), molecular layer deposition (MLD; a technique similar to ALD but differing in that molecules (typically 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 sequential chemistry. According to the invention, ALD is a preferred coating method.

[0072] When using ALD, the aforementioned mixed oxide coating can be prepared by feeding a first zinc-, other metal, or metalloid precursor into the ALD reactor chamber (in the form of a so-called "precursor pulse") to form an adsorbed atomic or molecular monolayer of zinc-, other metal, or metalloid on the surface of the particles. A second precursor (e.g., water) is then pulsed into the reactor and reacts with the first precursor, resulting in the formation of a zinc oxide monolayer, a metal oxide monolayer, or a metalloid monolayer, respectively, on the matrix surface. A subsequent purge pulse is followed by another first precursor pulse, thus initiating a new cycle of the same events (which is the ALD cycle).

[0073] When using MLD, organic coatings, polymers, and / or organic-inorganic hybrid coatings can be prepared by using MLD precursors, such as organic molecules containing bifunctional groups, such as diols, diamines, diisocyanates, dichlorides, and dialdehydes.

[0074] In most cases, the first reaction in a series of reactions will involve some functional group or free electron pair or radical at the surface to be coated, such as a hydroxyl group (-OH) or a primary or secondary amino group (-NH2 or -NHR, where R is, for example, an aliphatic group, such as an alkyl group). The individual reactions are advantageously carried out individually, and under conditions that all excess reagents and reaction products are substantially removed before subsequent reactions.

[0075] To prepare a mixed oxide coating in which the atomic ratio of zinc oxide to one or more other metal oxides and / or metalloids (in terms of the number of monolayers applied) is, for example, between at least about 1:1 and at most and including about 6:1, those skilled in the art will understand that for each ALD cycle (i.e., monolayer) of the other oxides, about one to about six ALD cycles of zinc oxide must also be deposited. For example, to form a 3:1 atomic (zinc oxide: other oxide) mixed oxide coating, a second precursor pulse may be performed after each of three zinc-containing precursor pulses to form three zinc oxide monolayers, followed by one pulse containing a precursor of the other metal and / or metalloids, and then a second precursor pulse to form one monolayer of the other metal and / or metalloid oxide. Alternatively, two other oxide monolayers or any other combination may be performed after six zinc oxide monolayers to provide an overall atomic ratio of about 3:1. In this respect, the order of the pulses used to produce the relevant oxides is not important, provided that the final atomic ratio is within the relevant range.

[0076] Metals and / or metalloids other than zinc that may be mentioned include alkali metals, alkaline earth metals, noble metals, transition metals, post-transition metals, and lanthanides. Metals and / or 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 metals and / or metalloids that may be mentioned include aluminum and silicon.

[0077] In this regard, the mixed oxide coating material preferably comprises one or both of alumina (Al2O3) and / or silicon dioxide (SiO2).

[0078] A method for preparing a plurality of coated particles according to the invention is provided, wherein the coated particles are prepared by vapor deposition by applying at least two metal oxide and / or metal-like oxide precursors to a solid core and / or a previously coated solid core to form a mixed oxide. The precursors used to form the metal oxide or metal-like oxide typically include oxygen precursors such as water, oxygen, ozone, and / or hydrogen peroxide; and metal and / or metal-like compounds, typically organometallic compounds or organometallic compounds.

[0079] Non-limiting examples of precursors are as follows: Zinc oxide precursors can be water and diC1-C5 alkyl zinc, such as diethylzinc. Aluminum oxide precursors can be water and triC1-C5 alkyl aluminum, such as trimethylaluminum. Silica (silicon dioxide) precursors can be water as an oxygen precursor and silanes, alkylsilanes, aminosilanes, and tetraethyl orthosilicate. Iron oxide precursors include oxygen, ozone, and water as an oxygen precursor; as well as diC1-C5 alkyl-iron, dicyclopropyl-iron, and FeCl3. It should be understood that those skilled in the art know which precursors are suitable for the purposes disclosed herein.

[0080] In ALD, the coating material layer can be applied at process temperatures ranging from about 20°C to about 800°C, or from about 40°C to about 200°C, for example, from about 40°C to about 150°C, or from about 50°C to about 100°C. The optimal process temperature depends on the reactivity of the substances used in the precursor and / or core (including bioactive agents, such as GLP1RA / salt) and / or the melting point of the core matrix. Lower temperatures, such as from about 30°C to about 100°C, are preferred. Specifically, in one embodiment of the method, temperatures of about 20°C to about 80°C, such as from about 30°C to about 70°C, such as from about 40°C to about 60°C, or such as about 50°C are used.

[0081] It has been found that when an ALD is applied at lower temperatures (e.g., from about 50°C to about 100°C) to form a coating containing zinc oxide (unlike other coating materials that form amorphous layers, such as alumina, titanium dioxide, and silica), the coating material is essentially crystalline.

[0082] As mentioned above and not limited by theory, because zinc oxide is crystalline, if zinc oxide is used alone as the coating material, it is understood that interfaces may form between adjacent crystals of zinc oxide deposited via ALD, through which a carrier system, medium, or solvent (e.g., an aqueous solvent system) in which zinc oxide is partially soluble can enter after being suspended. This is thought to potentially lead to excessively rapid dissolution for the intended reservoir-forming composition.

[0083] Furthermore, previous studies have shown that formulations containing certain active ingredients coated with zinc oxide have lower relative bioavailability than their uncoated counterparts when suspended in aqueous media. This lower relative bioavailability is attributed to the degradation of the active ingredients before they can be released into the systemic circulation. As mentioned above, water permeation through the crystallization interface within the zinc oxide coating is thought to lead to the hydrolysis of the active ingredients within the coated particles.

[0084] It has now been discovered that these problems can be mitigated by fabricating mixed oxide coatings as described herein. Specifically, by forming mixed oxide coatings, as described herein, consisting primarily but not entirely of zinc oxide, it has been possible to coat active ingredients with coatings that appear substantially amorphous, or composites between crystalline and amorphous materials, and / or coatings in which the ingress of injection media such as water can be reduced. In this regard, it appears that by employing the mixed oxide aspect of the present invention, either in a heterogeneous manner (where another oxide “fills” the gaps formed by the interface) or in a homogeneous manner (where a true composite of the mixed oxide material is formed during deposition in a manner that potentially avoids the interface in the first place), the presence of the aforementioned perceived interfaces can be reduced or completely avoided.

[0085] The vapor deposition reactor chamber used may optionally and / or preferably be a stationary vapor deposition reactor chamber. In the context of a vapor deposition reactor chamber, the term 'stationary' should be understood to mean that the reactor chamber remains stationary when used to perform vapor deposition techniques, excluding negligible movement and / or vibration, such as movement and / or vibration caused by, for example, related machinery.

[0086] Alternatively, for example, as described below, a so-called 'stop-flow' method can be employed. Using the stop-flow method, once the first precursor has been fed into the reactor chamber and before the first precursor is purged from the reaction chamber, the first precursor can be allowed to contact the core in the reactor chamber for a predetermined period of time (which can be considered as soaking time). During this predetermined period, preferably there is substantially no pumping that could cause gas flow and / or substantially no mechanical agitation of the core.

[0087] Employing a stop-flow method can increase coating uniformity by allowing each gas to conformally diffuse within a high aspect ratio matrix (such as powder). This benefit may be even more pronounced when using precursors with slow reactivity, as this gives the precursors more time to react on the surface. This can be particularly evident when depositing a mixed oxide coating according to the invention. For example, when depositing a mixed zinc oxide / alumina coating as described herein, a lower probability of reaction towards the matrix surface was found with zinc-containing precursors (such as diethylzinc (DEZ)) compared to, for example, aluminum-containing precursors (such as trimethylaluminum (TMA)).

[0088] In addition to producing coatings with good shell integrity and more controlled release profiles, this type of stop-flow method can also improve the ability to achieve specific coating compositions.

[0089] For example, when attempting to produce a coating containing a zinc to aluminum atomic ratio of 3:1 in the resulting shell as described above using a vapor phase technique, it was found that a ratio very close to 3:1 could be achieved using a stop-flow method compared to when depositing the material using a continuous flow of precursors.

[0090] Preferably and / or optionally, a 'multi-pulse' technique may also be used to feed a first precursor, a second precursor, or both precursors into the reactor chamber.

[0091] Using this type of multi-pulse technology, the precursor can be fed into the reactor chamber as multiple 'sub-pulses' rather than as a continuous pulse, where each sub-pulse lasts for 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). After each sub-pulse, the precursor can be allowed to contact the core in the reactor chamber for a predetermined period of time, such as about 1 second to 500 seconds, about 2 seconds to 250 seconds, about 3 seconds to 100 seconds, about 4 seconds to 50 seconds, or about 5 seconds to 10 seconds, for example, 9 seconds. Similarly, depending on the size and nature of the vapor deposition reactor, this time can be extended to several minutes (e.g., up to about 30 minutes). The introduction of the sub-pulses and the subsequent soaking time can be repeated a predetermined number of times in a single step, such as between about 5 to 1000 times, about 10 to 250 times, or about 20 to 50 times.

[0092] The formulations of the present invention that may be mentioned include those that are not prepared by such stop-flow methods.

[0093] The core may be coated with one or more separate discrete mixed oxide coating layers as defined herein. Preferably, more than one separate discrete mixed oxide layer, coating or shell (these terms are used interchangeably herein) is sequentially applied (i.e. 'applied separately') to the solid core containing GLP1RA.

[0094] The term 'single application' in 'separate layer, coating, or shell' means that a solid core can be coated with a first coating material layer formed by more than one (e.g., multiple or a group) cycles as described herein, each cycle producing a zinc oxide monolayer or other metal oxide and / or metal-like oxide (as the case may be) monolayer, and then the resulting coated core can be subjected to some form of deagglomeration process.

[0095] In other words, a 'vapor deposition (e.g., ALD) cycle' can be repeated several times to provide a 'vapor deposition (e.g., ALD) set', which can consist of, for example, 10, 25, or 100 cycles. However, after this set of cycles, the coated core can undergo some form of deagglomeration process, followed by another set of cycles.

[0096] This process can be repeated as desired, so the number of discrete coating material layers produced by multiple sets of cycles in the final coating corresponds to the number of these intermittent deagglomeration steps, wherein final mechanical deagglomeration is chosen before applying the final coating material layer (the last set of cycles).

[0097] The particles of the formulation may have between one and about 100 discrete oxide mixture (and, if appropriate, other coating materials as described below) layers, such as between two and about 50 discrete layers, such as between three and about 10 discrete layers, such as between three and about six discrete layers.

[0098] When referring to coated particles, the terms "deagglomeration" and "de-agglomeration" are used interchangeably, and the deagglomeration of coated particle aggregates is preferably accomplished by mechanical sieving techniques.

[0099] The coated core can undergo the aforementioned deagglomeration process internally without being removed from the device via a continuous process. Such a process involves forcing solid product agglomerates formed by coating the core through a sieve positioned within the reactor before a second and / or further coating, and is configured to deagglomerate any particle aggregates when forced onto the coated core by a forcing device applied within the reactor. This method may be continued multiple times as needed and / or as appropriate before applying the final coating as described herein.

[0100] Positioning the sieve within the reactor vessel means that coating can be applied via a continuous process that does not require removing particles from the reactor. Therefore, manual particle handling is unnecessary, as is external machinery to deagglomerate particles. This not only significantly reduces the time required for the coating process but also simplifies and reduces the risk of personnel handling hazardous (e.g., toxic) materials. It also enhances process repeatability by limiting manual labor and reducing the risk of contamination.

[0101] Alternatively, and / or preferably, the coated core can be removed from a coating apparatus such as an ALD reactor and then subjected to an external deagglomeration step, for example as described in International Patent Application WO 2014 / 187995. Such an external deagglomeration step may include stirring, such as ultrasonic treatment in a wet or dry state, or preferably may include subjecting the resulting solid product clumps discharged from the reactor to sieving, for example by forcing them through a sieve or sieve openings to deagglomerate the particles, as described below, and then returning the particles to the coating apparatus for the next coating step. Similarly, this process can be continued multiple times as needed and / or as appropriate before applying the final coating.

[0102] In the external deagglomeration process, deagglomeration can be alternatively (and / or in lieu of the above processes) achieved 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 is performed on wet particles, the deagglomerated particles should be dried (as described above regarding the core) before the next coating step.

[0103] However, preferably, in such external processes, the deagglomeration step includes one or more screening steps, which may include jet screening, manual screening, vibrating screen shaking, horizontal screen shaking, tapping screening, or (preferably) acoustic screening or similar methods as described below, including any combination of these screening steps. Suitable manufacturers of acoustic screens include Advantech Manufacturing, Endecott, and Tsutsui.

[0104] Vibratory sieving technology can involve a device that uses vibration to force solid product agglomerates formed by coating the core through a sieve located inside or (preferably) outside the reactor, and configured to deagglomerate any particle aggregates under the vibrational force of the coated core before applying a second and / or additional coating material layer. This method is repeated multiple times as needed and / or as appropriate before applying the final coating material layer.

[0105] The device for applying vibration includes a vibrating motor coupled to the screen. The vibrating motor is configured to vibrate and / or rotate when power is supplied to it. For example, the vibrating motor may be a piezoelectric vibrating motor comprising a piezoelectric material that changes shape due to the inverse piezoelectric effect when an electric field is applied. The change in shape of the piezoelectric material causes acoustic or ultrasonic vibration of the piezoelectric vibrating motor.

[0106] Alternatively, the vibration motor can be an eccentric rotating mass (ERM) vibration motor, which comprises a mass that rotates when power is supplied to the motor. The mass deviates from the axis of rotation, causing the motor to become unbalanced and vibrate and / or rotate due to the rotation of the mass. Furthermore, the ERM vibration motor can comprise multiple masses positioned at different locations relative to the motor. For example, the ERM vibration motor can comprise a top mass and a bottom mass, each positioned at opposite ends of the motor. By changing each mass and its angle relative to another mass, the vibration and / or rotation of the ERM vibration motor can be altered.

[0107] The vibratory motor is coupled to the screen in such a way that when power is supplied to it, the vibration and / or rotation of the motor are transferred to the screen.

[0108] The screen and vibrating motor can be suspended from a mounting (e.g., a frame that can be positioned on the floor) via a suspension device, allowing the screen and motor to vibrate freely relative to the mounting without the vibration being substantially transferred to or damped by the mounting. This allows the vibrating motor and screen to vibrate and / or rotate unimpeded and also reduces noise generated in vibrating screening methods. The suspension device may include one or more springs or bellows (i.e., air cushions or equivalent damping devices) that couple the screen and / or motor to the mounting. Manufacturers of vibrating screens or sieves suitable for such processes include, for example, Russell Finex, SWECO, Filtra Vibracion, VibraScreener, Gough Engineering, and Farley Greene.

[0109] Preferably, the vibrating screening technology further includes controlling a vibration probe coupled to the screen. The vibration probe can be controlled to cause the screen to vibrate at a frequency different from the vibration frequency caused by the vibrating motor. Preferably, the vibration probe causes the screen to vibrate at a higher frequency than the vibration caused by the vibrating motor, and more preferably, the frequency is in the ultrasonic range.

[0110] By using a vibrating probe to provide additional vibration to the screen, the occurrence of screen blockage is reduced, the possibility of screen overload is reduced, and the amount of time required to clean the screen holes is reduced.

[0111] Preferably, the aforementioned vibrating sieving technology includes sieving the coated particles at a throughput of at least 1 g / min. More preferably, the vibrating sieving technology includes sieving the coated particles at a throughput of 4 g / min or more.

[0112] The throughput depends on the sieve aperture area, sieve aperture size, particle size, particle viscosity, and static properties of the particles. By combining some of these characteristics, much higher throughputs can be achieved. Therefore, vibrating screening technology can more preferably include screening coated particles at a throughput of up to 1 kg / min or even higher.

[0113] Each of the above-mentioned throughputs represents a significant improvement over the use of known mechanical sieving or screening techniques. For example, it has been found that sonic sieving involves sieving over a 15-minute time period, with a 15-minute cooling time in between, which is necessary to protect the equipment. To sieve 20g of coated particles, nine sets of 15-minute active sieving times are required, for a total time (including cooling) of 255 minutes. In contrast, by using the above-described vibrating sieving technique, 20g of coated particles can be sieved continuously for at most 20 minutes, or more preferably only 5 minutes or less.

[0114] The sieve aperture size can be determined such that the ratio of particle size to sieve aperture size after sieving or sonic sieving is about 1:>1, preferably about 1:2, and optionally about 1:4. The sieve aperture size can range from about 20 μm to about 100 μm, preferably from about 20 μm to about 60 μm.

[0115] Suitable sieve openings may include perforated plates, microporous plates, meshes, diamond mesh, fine wires, polymers, or wires (woven wire sieves), but are preferably formed of metal (such as stainless steel).

[0116] Surprisingly, the effect of using stainless steel mesh in vibrating screening technology on particle coating is as mild as that of using softer polymer screens in mechanical screening technologies such as sonic screening.

[0117] Furthermore, a known problem with sieving powders is the potential danger of generating static electricity. Steel sieves have the advantage of removing static electricity from powders, while polymer sieves do not, and these polymer sieves must be used in acoustic sieves.

[0118] Furthermore, the sieve aperture size of known acoustic sieves is limited to approximately 100 μm because the acoustic waves travel through the sieve apertures rather than vibrate them. For vibratory sieving technology, this limitation does not exist because it does not rely on acoustic waves generating vibrations within the sieve. Therefore, the vibratory sieving technology described herein allows for the sieving of larger particles compared to alternative mechanical sieving techniques.

[0119] If (e.g., vibrating) screen is located outside the reactor (i.e., outside), the method for preparing the coated core of the formulation of the present invention includes discharging the coated particles from the vapor deposition reactor before stirring the coated particles, then reintroducing the deagglomerated coated particles into the vapor deposition reactor, and then applying at least one additional coating material layer onto the reintroduced particles.

[0120] As described above, it has been found that applying a separate coating layer after external deagglomeration produces a visible and identifiable interface as a region of higher electron permeability, which can be observed by analyzing the coated particles according to the invention and by, for example, TEM. In this respect, the thickness of the layer between the interfaces directly corresponds to the number of cycles in each series performed within the ALD reactor and between the various external agitation steps.

[0121] Because coating occurs at the atomic level in the ALD coating method, such clear physical interfaces are often more difficult to observe.

[0122] Unrestricted by theory, it is believed that removing coated particles from the vacuum conditions of an ALD reactor and exposing the newly coated surface to the atmosphere will lead to structural rearrangement due to relaxation and reconstruction of the outermost atomic layer. Such methods are thought to involve the rearrangement of surface (and near-surface) atoms, driven by a thermodynamic tendency to reduce surface free energy.

[0123] Furthermore, surface adsorption of substances, such as hydrocarbons that are always present in the air, can cause this phenomenon due to the coating reaction formed by hydrocarbons and atmospheric oxygen. Surface modification can also cause this phenomenon. Therefore, if such interfaces are chemically analyzed, they may contain trace amounts of contaminants or core materials, such as active ingredients that form part of the core, which are not derived from the coating process, such as ALD.

[0124] Whether carried out inside or outside the reactor, particle aggregates are preferably broken up by a forced device that forces them through a sieve, thereby separating the aggregates into individual particles or aggregates of a desired and predetermined size (and thus achieving deagglomeration). In the latter case, in some situations, the individual primary particle size is so small (i.e., <1 μm) that 'complete' deagglomeration (i.e., where the aggregates break down into individual particles) is not possible. Instead, deagglomeration is achieved by breaking down larger aggregates into smaller aggregates of secondary particles of a desired size, specified by the size of the sieve apertures. The smaller aggregates are then coated using gas-phase techniques to form fully coated 'particles' in the form of small aggregate particles. In this way, the term 'particle' in the context of this invention, referring to both deagglomerated and coated particles, refers to individual (primary) particles and aggregates (secondary) particles of a desired size.

[0125] In any case, the desired particle size (whether individual particles or aggregates of the desired size) remains unchanged, and furthermore, after such deagglomeration by a sieving device, applying a gas-phase coating mechanism to the particles means forming a complete coating on the particles, thereby forming fully coated particles (individual particles or aggregates of the desired size).

[0126] Whether carried out inside or outside the reactor, the repeated coating and deagglomeration process described above can be performed at least once, preferably twice, more preferably three times, such as four times, including five times, more specifically six times, such as seven times, and not more than about 100 times, such as not more than about 50 times, such as not more than about 40 times, including not more than about 30 times, such as between two and 20 times, such as between three and 15 times, such as 10 times, such as nine or eight times, more preferably six or seven times, and especially four or five times.

[0127] Whether carried out inside or outside the reactor, it is preferred that at least one screening step be performed, and more preferably, the step preferably includes a vibratory screening step as described above. More preferably, at least the final screening step includes a vibratory screening step performed before the application of the final coating material layer (circulation group). However, it is even more preferred that more than one (including each) of the screening steps includes the vibratory screening technique, procedure, or method as described herein.

[0128] Repeating these steps is preferred to make it more advantageous to increase the throughput of any vibrating screening technology.

[0129] The formulations mentioned in this invention include those not prepared by processes including vibratory sieving techniques.

[0130] The total thickness of the coating (meaning all the individual layers / coatings / shells) will average between approximately 0.5 nm and approximately 2 μm.

[0131] The minimum thickness of each individual layer / coating / shell will be in the range of approximately 0.1 nm (including approximately 0.5 nm, such as approximately 0.75 nm, or approximately 1 nm).

[0132] The maximum thickness of each individual layer / coating / shell will depend on the size of the core (initially), then on the size of the core to which coating has been previously applied, and may average approximately 1 percent of the average diameter of the core or the core to which coating has been previously applied (i.e., the average diameter based on weight, quantity, or volume).

[0133] Preferably, for particles with an average diameter between about 100 nm and about 1 μm, the total coating thickness should be between about 1 nm and about 5 nm on average; for particles with an average diameter between about 1 μm and about 20 μm, the coating thickness should be between about 1 nm and about 10 nm on average; and for particles with an average diameter between about 20 μm and about 700 μm, the coating thickness should be between about 1 nm and about 100 nm on average.

[0134] In this respect, the preferred average diameter of the coated core of the formulation of the present invention, based on weight, quantity or volume, is preferably between about 50 nm (e.g., about 100 nm, such as about 250 nm) and about 30 μm, such as between about 500 nm and about 100 μm, more specifically between about 1 μm (such as about 5 μm, including about 7 μm, about 9 μm, about 10 μm or about 15 μm) and about 50 μm, such as about 25 μm, such as about 20 μm.

[0135] It has been found that applying a coating / shell followed by one or more deagglomeration steps, such as sonication, can produce abrasion, pinholes, fractures, gaps, cracks, and / or voids (hereinafter referred to as 'cracks') in the layers / coating. This is because the coated particles essentially "glued" or "adheded" together more tightly after a thicker coating is applied. Once deagglomeration occurs, this can expose the core containing the bioactive ingredients to the environment.

[0136] Since the aim is to provide the particles in the suspension prior to administration to the patient, it is necessary to provide primary particles that have been deagglomerated and free of pinholes or cracks in the coating. Such cracks will result in an undesirable initial spike (sudden increase) in the concentration of the active ingredient in the plasma directly after administration.

[0137] It has been found that by performing one or more of the deagglomeration steps described herein, a significant reduction in pinholes, gaps, or cracks is produced in the final coating material layer, resulting in particles that are not only completely coated by the layer / coating, but also coated in a manner that allows the particles to be easily deagglomerated (e.g., using non-erosive techniques such as eddy currents), without damaging the coating material layer formed before and / or during drug preparation.

[0138] In this regard, the mixed oxide coating typically completely surrounds, encapsulates, and / or encapsulates the solid containing the active ingredient. In this way, the risk of initial drug concentration bursts due to direct contact between the drug and a solvent in which the relevant active ingredient is soluble is minimized. This can include not only bodily fluids but also any medium in which such coated particles can be suspended prior to injection.

[0139] Therefore, in yet another embodiment of the invention, particles as disclosed above are provided, wherein the coating surrounding, encapsulating, and / or encapsulating the core covers at least about 50%, such as at least about 65%, including at least about 75%, such as at least about 80%, more specifically at least about 90%, such as at least about 91%, such as at least about 92%, such as at least about 93%, such as at least about 94%, such as at least about 95%, such as at least about 96%, such as at least about 97%, such as at least about 98%, such as at least about 99%, such as about or about 100% of the surface of the solid core, such that the coating substantially completely surrounds, encapsulates, and / or encapsulates the core.

[0140] As used herein, the term 'substantially completely surrounds, wraps and / or encapsulates the core' means covering at least about 98% or at least about 99% of the solid core surface.

[0141] In an alternative, the method described herein may result in deagglomerated coated particles that are essentially free of active ingredients that can pass through the cracks and be released in an uncontrolled manner.

[0142] Although some minor cracks may appear in the coating without affecting its basic function in controlled release, in another embodiment, particles as disclosed above are provided, wherein at least about 90% of the particles are free of cracks in the coating surrounding, encapsulating, and / or encapsulating the core. In one embodiment, at least about 91%, such as at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% of the particles are free of the cracks.

[0143] Alternatively, “substantially free of said cracks” in the coating also means that less than about 1% of the surface of the coated particles contains abrasion, pinholes, breaks, gaps, cracks and / or voids through which the active ingredient may be potentially exposed (e.g., exposed to the environment).

[0144] The coating material layers can together have a substantially uniform thickness over the surface area of ​​the particles. The term 'substantially uniform' thickness means, as measured by TEM, that at least about 10%, such as about 25%, for example about 50%, of the coated particles present in the formulation of the present invention have a thickness variation of no more than about ±20%, including ±50%, of the average thickness.

[0145] In addition to the basic mixed oxide coating used in the formulations of this invention, other coating materials, which may be pharmaceutically acceptable and substantially non-toxic, may be applied between individual mixed oxide coatings (e.g., between individual deagglomeration steps) and / or concurrently with the application of the mixed oxide coating herein. Such materials may comprise multiple layers (or composites) of the mixed oxide and one or more different inorganic or organic materials to modify the properties of the layers.

[0146] Other coating materials may comprise organic or polymeric materials such as polyamides, polyimides, polyureas, polyurethanes, polythioureas, polyesters, or polyimides. Additional coating materials may also comprise hybrid materials (such as combinations of organic and inorganic materials), including materials combining a metal or another element with an alcohol, carboxylic acid, amine, or nitrile. Such additional organic coatings, polymer coatings, and / or hybrid organic-inorganic coatings are preferably applied using coating techniques incorporating MLDs as described above. Such polymer coatings may be polyimides, polyazomethine, polyureas, polyamides, nylon, metal cones, aluminum cones, titanium cones, zinc cones, metal-organic framework polymers, carbon oxides, and hybrid nanolaminated materials.

[0147] However, the preferred coating material contains inorganic materials.

[0148] Other inorganic coating materials (besides zinc oxide) may contain other compounds of metals and / or metalloids, such as oxides, nitrides, sulfides, selenides, carbonates, and other ternary compounds. Metals and metalloids, hydroxides, and especially oxides are preferred, particularly metal oxides.

[0149] Additionally, oxides of elements other than zinc, aluminum, or silicon may be mentioned, including alkali metals, alkaline earth metals, noble metals, transition metals, post-transition metals, and lanthanides. 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.

[0150] Other coating materials that can be mentioned therefore include those containing titanium dioxide (TiO2) and iron oxide (Fe). x O y Coating materials such as 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).

[0151] Preferably, before and / or after applying the mixed oxide layer as described herein to the core, an additional inorganic material comprising one or more single metal oxide layers (e.g., titanium oxide, silicon oxide, or preferably aluminum oxide) is applied to the core.

[0152] Although the plurality of mixed oxide-coated particles according to the invention have substantially no active ingredient in the applied coating through the aforementioned cracks that are potentially exposed (e.g., exposed to the outside), two additional optional steps may be applied to the plurality of coated particles before further pharmaceutical formulation processing.

[0153] The first optional step may be to apply a final outer coating layer after the final deagglomeration step as described above, the thickness of which must be thinner than the previously applied individual layer / coating / shell (or 'subshell').

[0154] Therefore, the thickness may, on average, not exceed about 0.7 times (e.g., about 0.6 times) the thickness of the widest previously applied subshell. Alternatively, the thickness may, on average, not exceed about 0.7 times (e.g., about 0.6 times) the thickness of the last applied subshell, and / or may, on average, not exceed about 0.7 times (e.g., about 0.6 times) the average thickness of all previously applied subshells. For particles up to about 20 μm, the average thickness may range from about 0.3 nm to about 10 nm. For larger particles, the average thickness may not exceed about 1 / 1000 of the average diameter of the coated particles based on weight, number, or volume.

[0155] The function of the sealing shell is to provide a 'sealing' outer coating layer on the particles, which covers those cracks, thereby producing particles that are not only completely covered by the sealing shell, but are also covered in a way that allows the particles to easily deagglomerate (e.g., using non-erosive techniques such as vortexing), so as not to destroy the subshells that have already formed underneath before and / or during drug preparation.

[0156] For the reasons described herein, it is preferable that the sealing shell does not contain zinc oxide. On the other hand, the sealing shell may contain silicon dioxide, or more preferably aluminum oxide.

[0157] The second optional step may include ensuring that a small number of remaining particles with broken and / or cracked shells / coatings are treated, wherein all particles are suspended in a solvent in which their active ingredient is soluble (e.g., with a solubility of at least about 0.1 mg / mL), but the least soluble substance in the mixed oxide coating is insoluble (e.g., with a solubility of no more than about 0.1 μg / mL), and then the solid particles are separated from the solvent by, for example, centrifugation, sedimentation, flocculation and / or filtration, resulting in the retention of predominantly intact particles.

[0158] The optional steps mentioned above provide a means to potentially further reduce the likelihood of (potentially) undesirable initial peaks (burst releases) in the plasma concentration of the active ingredient, as discussed above.

[0159] At the end of the method, the coated particles can be dried using one or more of the techniques described above for drying the core. Drying can be carried out in the absence or in the presence of one or more pharmaceutically acceptable excipients (e.g., sugars or sugar alcohols).

[0160] Alternatively, at the end of the method, the separated particles may be resuspended in a solvent (e.g., water, with or without one or more pharmaceutically acceptable excipients as defined herein) for subsequent storage and / or administration to a patient.

[0161] Before applying the first coating layer or between consecutive coatings, the core and / or partially coated particles may undergo one or more alternative and / or preparatory surface treatments. In this regard, one or more intermediate layers comprising different materials (i.e., materials other than inorganic materials) may be applied to the relevant surface, for example to protect the core or partially coated particles from undesirable reactions with the precursor during the coating step / deposition process, thereby improving coating efficiency or reducing agglomeration.

[0162] For example, the intermediate layer may contain one or more surfactants to reduce the aggregation of particles to be coated and to provide a hydrophilic surface suitable for subsequent coating. Suitable surfactants in this regard include well-known nonionic, anionic, cationic, or amphoteric surfactants, such as the Tween series, for example, Tween 80. Alternatively, the core may be pre-treated with a surface treatment if the active ingredient used as part of (or as the core) readily reacts with one or more precursor compounds that may be present in the gas phase during the coating (e.g., ALD) process.

[0163] The application of this type of "intermediate" layer / surface treatment can alternatively be achieved through liquid-phase uncoating techniques, followed by freeze-drying, spray drying, or other drying methods to provide particles with a surface layer on which a coating material can then be applied.

[0164] The outer surface of the particles of the formulations of the present invention can also be derivatized or functionalized, for example by attaching one or more compounds or portions to the outer surface of the final coating material layer, such as compounds or portions that enhance the targeted delivery of the particles in a patient in whom the nanoparticles are administered. Such compounds can be organic molecules (such as PEG) polymers, antibodies or antibody fragments, or receptor-binding proteins or peptides, etc.

[0165] Alternatively, the moiety can be an anchoring group, such as a moiety containing a silane functional group (see, for example, Herrera et al., *Journal of Materials Chemistry*, 18, 3650 (2008) and US 8,097,742). Another compound, such as the desired targeting compound, can be attached to such an anchoring group by means of covalent or non-covalent bonding (including hydrogen bonding or van der Waals bonding) or a combination thereof.

[0166] The presence of such anchoring groups can provide a universal tool for targeted delivery to specific sites in the body. Alternatively, the use of compounds such as PEG may result in particles circulating in the bloodstream for a longer period, thus ensuring that particles do not accumulate in the liver or spleen (the body's natural mechanism for eliminating particles, which can prevent delivery to diseased tissues).

[0167] Cores coated with a mixture of oxides, whether as a single layer, discrete layer, coating or shell or other form, as defined herein, are referred to hereinafter as “coated particles of the formulation of the present invention”.

[0168] According to another aspect of the invention, the coated particles of the formulation of the invention comprise a plurality of particles, said plurality of particles:

[0169] (a) Having an average diameter based on weight, quantity, or volume between 10 nm and approximately 700 μm; and

[0170] (b) Containing a core, the core comprising one or more GLP1RAs or pharmaceutically acceptable salts thereof, the core being at least partially coated with an inorganic material coating comprising a mixture of the following:

[0171] (i) Zinc oxide (ZnO); and

[0172] (ii) one or more other metal oxides and / or metal-like oxides.

[0173] The atomic ratio ((i):(ii)) is at least about 1:10 and at most and includes about 10:1.

[0174] 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 to the aspects and / or preferences of the coated particles of the formulations of the present invention described above. Furthermore, to avoid ambiguity, these aspects, preferences, and features, individually or in combination, are hereby incorporated into this aspect of the invention by reference.

[0175] The pharmaceutical (or veterinary) formulations of the present invention may include different types of particles, such as particles containing different functionalizations (as described above), particles of different sizes, and / or mixed oxide coating material layers of different thicknesses, 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., altered or prolonged) over a specific time period.

[0176] The formulations of the present invention can be administered systemically, for example, by intravenous or intra-arterial infusion (including via intravascular or other perivascular devices / dosage forms (e.g., stents)), intraosseous, intracerebral, intraventricular, intrasynovial, intrasheath, intralesional, intracranial, intratumoral, intradermal, intradermal, or most preferably intramuscular or subcutaneous injection in pharmaceutically (or veterinary) dosage forms.

[0177] The preparation of formulations of the present invention comprises incorporating coated particles, as described herein, into a suitable pharmaceutically acceptable (including physiologically acceptable) carrier system, and can be achieved by appropriately considering the intended route of administration and standard pharmaceutical practice. Therefore, suitable excipients (including physiologically acceptable injectable excipients, such as those mentioned herein that are physiologically acceptable for intramuscular injection or, more preferably, physiologically acceptable for subcutaneous injection) should be chemically inert to the active agent employed and have no harmful side effects or toxicity under the conditions of use. Such pharmaceutically acceptable carriers can also impart immediate or modified release of the active agent from the particles of the formulations of the present invention.

[0178] Therefore, a pharmaceutically acceptable carrier system according to the invention can be an oil-based or oil-based system. The carrier system can thus comprise one or more pharmaceutically or veterinarily acceptable liquid lipids, said liquid lipids may include fixed oils such as monoglycerides, diglycerides, or triglycerides, including Miglyol. TM (e.g., 812N), propylene glycol dioctyl decanoate (Miglyol 840, C8 / C10 ester), trioctyl ester (Miglyol oil), Gelucire TM 43 / 01, Kollisolv TM GTA, Labrafil TMThe carrier system may also comprise polysorbates (such as polysorbate 20, polysorbate 60, polysorbate 80), glycols (such as propylene glycol, polyethylene glycol, polyethylene glycol 300, polyethylene glycol 400, 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 their polyoxyethyleneized versions (e.g., sorbitan trioleate, propylene glycol monolaurate 90, Capryol). TM PGMC, PEG-60 hydrogenated castor oil, and polyoxyethylene 35 castor oil are all suitable carrier systems. Non-ester oils, such as ethyl lactate and perfluorohexyl octane (F6H8), can also be used. More preferred carrier systems include monoglycerides, diglycerides, and / or triglycerides, with medium-chain triglycerides, such as alkyl-chain triglycerides (e.g., C6-C...). 12 Alkyl chain triglycerides).

[0179] The sterile aqueous suspension of the 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 also contain other aqueous excipients, such as Ringer's solution, and may also include polar cosolvents (e.g., ethanol, glycerol, propylene glycol, 1,3-butanediol, polyethylene glycol of various molecular weights, and tetraethylene glycol); thickeners or viscous agents (e.g., carboxymethyl cellulose, microcrystalline cellulose, hydroxypropyl methyl cellulose, hydroxyethyl cellulose, ethyl hydroxyethyl cellulose, sodium glycolate starch, poloxamer (e.g., poloxamer 407), polyvinylpyrrolidone, cyclodextrin (e.g., hydroxypropyl-β-cyclodextrin), polyvinylpyrrolidone, and polyethylene glycol of various molecular weights); surfactants / wetting agents to obtain a homogeneous suspension (e.g., sorbitol esters, sodium lauryl sulfate; glyceryl monoesters, polyoxyethylene esters, polyoxyethylene alkyl ethers, polyoxyglycerol esters, and preferably Tween (polysorbate), such as Tween 80 and Tween 20). Preferred components include isotonic adjusters (e.g., sodium lactate, glucose, and especially sodium chloride); pH adjusters and / or buffers (e.g., citric acid, sodium citrate, and especially phosphate buffers (such as disodium hydrogen phosphate dihydrate, sodium acid phosphate, sodium dihydrogen phosphate monohydrate, and combinations thereof), which can be used in combination with standard inorganic acids and standard inorganic bases (such as hydrochloric acid and sodium hydroxide); and other components such as mannitol, croscarmellose sodium, and hyaluronic acid.

[0180] The formulations of the present invention can be further formulated into an injectable suspension of coated particles having a uniform size distribution and capable of forming (or reforming upon appropriate agitation) a stable suspension (i.e., without sedimentation) within the injection solution, allowing it to be injected through a needle. In this respect, the formulations of the present invention may contain inactive ingredients that prevent premature 'caking' (i.e., formation of solid or semi-solid, non-dispersible residues) or gelling (e.g., formation of hydrogels), meaning the formulation is sufficiently viscous to prevent sedimentation, resulting in an "uneven" suspension and thus a risk of insufficient or excessive dosage of the active ingredient, or at least can be redispersed before application to form a sufficiently uniform dispersion.

[0181] In view of the above requirements, the formulations of the present invention comprising an aqueous carrier system in which coated particles are suspended may further comprise:

[0182] i. Physiologically acceptable injectable (e.g., intramuscular and / or more preferably subcutaneous) compounds that react with zinc and, in doing so, reduce (e.g., substantially prevent) the reaction of zinc with water to form zinc hydroxide. Such compounds may, for example, contain compounds that provide a source of counterions when dissolved in an aqueous carrier, said counterions being capable of forming compounds (e.g., salts) with zinc that are substantially insoluble in said aqueous medium (at any given temperature, pressure, and pH); and

[0183] ii. A physiologically acceptable injectable (e.g., intramuscular and / or more preferably subcutaneous) buffer system, wherein the buffer system:

[0184] 1. It basically cannot form compounds (e.g., salts) with zinc; or

[0185] 2. Contains a counterion source that is capable of forming a compound (e.g., a salt) with zinc that is more soluble in the aqueous medium than zinc hydroxide (Zn(OH)2).

[0186] It has been found that when a medium- to high concentration of at least one GLP1RA (e.g., at least about 10 mg per milliliter of carrier, such as between about 25 mg and about 200 mg per milliliter of carrier, such as between about 30 mg / mL and about 150 mg / mL, such as between about 40 mg / mL and about 100 mg / mL, such as about 50 mg / mL) is present in an aqueous medium, viscous aggregates and / or hydrogels may form, which tend to clog needles when attempting to inject the formulation, for example, subcutaneously or intramuscularly.

[0187] It is believed that the accidental formation of aggregates and / or hydrogels is due to the reaction of zinc in the coating with water to form zinc hydroxide, resulting in an unstable and non-injectable suspension.

[0188] It has been found that this unforeseen problem can be solved by adding a compound that competes with water for the reaction with zinc. Thus, this compound is able to react with zinc and, in doing so, reduces and / or substantially prevents (e.g., prevents up to about 75%, such as about 80%, including up to about 90%, such as about 95%, and even about 99%) the reaction of zinc with water to form zinc hydroxide.

[0189] Such compounds can, for example, provide a source of counterions for aqueous carrier systems. When the source of counterions is included (e.g., dissolved) in the latter, it can form compounds (e.g., salts) with zinc to prevent the aforementioned reactions of free zinc with water. These counterions can be achieved, for example, by complexing with zinc in some way, and / or by being substantially insoluble in water and / or precipitating from aqueous solutions. The competing reaction can occur on or near the surface of the coated particles. Regardless of the mechanism involved, in the process of competing with water for the reaction with zinc, further reactions of zinc with water (and thus the formation of gels) are reduced and / or prevented.

[0190] The term 'substantially insoluble' in aqueous media (such as pure water) includes zinc compounds (e.g., salts) that are slightly soluble in such media at atmospheric pressure (e.g., about 1 bar), at room temperature (e.g., about 21°C), and at neutral pH (e.g., pH between about 5 and about 9, such as between about 6 and about 8.5, such as between about 7 and about 8 (e.g., about 7.4)), such as those with solubility less than about 33.3 mg / mL, such as less than about 25 mg / mL, including less than about 20 mg / mL, and especially less than about 10 mg / mL, such as less than about 5 mg / mL, as low as less than about 1 mg / mL, and including those zinc compounds with solubility less than about 0.1 mg / mL.

[0191] Preferably, at any given temperature, pressure, and pH, the solubility of zinc-related compounds (e.g., salts) in aqueous media (e.g., pure water) is lower than that of zinc hydroxide (Zn(OH)2) (e.g., at least about 10%, such as at least about 5%).

[0192] In this regard, suitable counterions with the above-mentioned properties include aspartate counterions, tartrate counterions, maleate counterions, fumarate counterions, malate counterions, benzoate counterions, and preferably phosphate counterions. Suitable sources of such counterions include substances capable of forming buffers based on aspartate, tartrate, maleate, fumarate, malate, benzoate, and / or phosphate, such as: aspartic acid, aspartate (e.g., sodium aspartate) and their hydrates, and mixtures thereof; tartaric acid, tartrate (sodium tartrate) and their hydrates (e.g., dihydrates), and mixtures thereof; maleic acid, maleate (e.g., sodium maleate) and their hydrates, and mixtures thereof; fumaric acid, fumarate (e.g., monosodium fumarate) and their hydrates, and mixtures thereof; malic acid, malate and their hydrates, and mixtures thereof; benzoic acid, benzoate (e.g., sodium benzoate) and their hydrates, and mixtures thereof; and more specifically, phosphate buffers (such as phosphate, disodium hydrogen phosphate dihydrate, sodium acid phosphate, sodium dihydrogen phosphate monohydrate, and combinations thereof).

[0193] Alternative sources of phosphate counterions include salts such as sodium phosphate, potassium phosphate, and calcium phosphate. Sources may also include organophosphates (e.g., glycerophosphate, sodium glycerophosphate, and potassium glycerophosphate), as well as pyrophosphates and polyphosphates. Alternative sources of tartrate counterions include potassium tartrate, diethyl tartrate, and disodium tartrate. Alternative sources of benzoate counterions include sodium benzoate, benzyl benzoate, denatumium benzoate, and potassium benzoate.

[0194] The phosphate counterions in the aqueous media of the formulations of the present invention can have suitable concentrations ranging from about 1 mM, such as from about 2 mM to about 50 mM, including about 40 mM, such as from about 3 mM to about 35 mM, for example, between about 4 mM (e.g., about 5 mM) and about 30 mM (e.g., about 25 mM, including about 20 mM, about 15 mM and about 10 mM). (In, for example, subcutaneously injectable compositions, the recommended upper limit of the concentration of phosphate buffer is 10 mM; see Usach et al., Adv. Ther. 36, 2986 (2019)).

[0195] Although the counterion sources mentioned above, capable of forming zinc compounds (e.g., salts), could prevent the gelling problem described here, it has been unexpectedly found that this can cause pH changes in the resulting formulations (especially over time, e.g., during storage). This is thought to be due to the reaction of the aforementioned counterions (e.g., phosphate ions) with zinc, depleting the buffering capacity of the relevant counterion sources (e.g., buffer solutions).

[0196] This problem can be addressed by adding a physiologically acceptable injectable (e.g., intramuscular or, more preferably, subcutaneous) buffer system, wherein the buffer system:

[0197] • Contains 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,

[0198] • It can basically not form salts with zinc.

[0199] The presence of such buffer solutions, when combined with the aforementioned other counterion sources, counteracts the effects of buffer depletion on the latter's buffering capacity, and thus helps maintain a constant (e.g., physiologically acceptable) pH within the formulations of the present invention. By "maintaining a constant pH" within the formulations of the present invention, it has been found that, during storage, the average pH change is less than ±20%, such as less than ±10%, including less than ±5%, compared to the pH measured immediately after the preparation of the formulations of the present invention.

[0200] Buffers containing counterion sources of salts that can form with zinc in aqueous media more readily soluble 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., glutamate, monosodium glutamate, and combinations thereof), succinate buffers (e.g., succinic acid, sodium succinate, and combinations thereof), α-ketoglutarate buffers (α-ketoglutarate, α-ketoglutarate, and combinations thereof), ascorbate buffers (e.g., ascorbic acid, sodium ascorbate, and combinations thereof), bicarbonate buffers (e.g., carbonic acid, sodium bicarbonate, and combinations thereof), ammonium salt buffers (e.g., ammonium chloride, ammonium hydroxide, and combinations thereof), glycine buffers (e.g., glycine, sodium glycinate, and combinations thereof), or any combination of the above substances.

[0201] Buffers that are generally non-salting with zinc include histidine, diethanolamine (e.g., diethanolamine, magnesium chloride hexahydrate, and combinations thereof), or most preferably tromethamine ('Tris' or 'Trizma') buffers.

[0202] All of the above-described buffer solutions can be used alone or in combination with standard inorganic acids and bases (such as hydrochloric acid and sodium hydroxide) that can be used to adjust the pH. The preferred pH of the formulations of the present invention can be in the pH range of about pH 3 to about pH 10, such as about pH 4 to about pH 9, including about pH 5 to about pH 8.

[0203] Suitable concentrations of such buffers (e.g., tromethamine buffer) are in the range of about 0.1 mM (e.g., about 5 mM, including about 10 mM) to about 200 mM, such as about 25 mM to about 175 mM, for example between about 50 mM and about 150 mM, including between about 75 mM and about 125 mM (e.g., about 100 mM).

[0204] The formulations of the present invention may also include a carrier system, which is a mixture of one or more oil and water-based systems, such as an emulsion, like an oil-in-water emulsion or an oil-in-water emulsion.

[0205] In (specifically oil-based) carriers, suitable dispersants or wetting agents (e.g., Tween, such as Tween 80) and / or suspending agents may be used.

[0206] Therefore, formulations can be stored under normal storage conditions and maintain their physical and / or chemical integrity. The phrase 'maintain physical and chemical integrity' essentially refers to both chemical and physical stability.

[0207] 'Chemical stability' includes the ability of any formulation of the present invention to be stored under normal storage conditions (with or without suitable pharmaceutical packaging), the degree of chemical (including stereochemical) degradation or decomposition of any active ingredient (especially any GLP1RA) and / or inert excipients and / or the aforementioned pH changes being unremarkable.

[0208] 'Physical stability' includes the ability of any formulation of the present invention to be stored under normal storage conditions (with or without appropriate pharmaceutical packaging) with no significant degree of physical transformation (sedimentation as described above), or with no significant change in the properties and / or integrity of the coated particles, such as the coating itself or the active ingredient (including dissolution, solvation, solid-state phase transition, etc.).

[0209] Examples of 'normal storage conditions' for the formulations of the present invention include a temperature between about -50°C and about +80°C (preferably between about -25°C and about +75°C, such as about 50°C), and / or a pressure between about 0.1 bar and about 2 bar (preferably atmospheric pressure), and / or exposure to about 460 lux of UV / visible light, and / or a relative humidity between about 5% and about 95% (preferably about 10% to about 40%) for a prolonged period of time (i.e., greater than or equal to about twelve months, such as about six months).

[0210] Under such conditions, it can be found that the formulations of the present invention are suitably chemically and / or physically degraded / decomposed by less than about 15%, more preferably less than about 10%, and especially less than about 5%. Those skilled in the art will understand that the upper and lower limits of temperature and pressure mentioned above represent extreme values ​​of normal storage conditions, and certain combinations of these extreme values ​​will not be experienced during normal storage (e.g., at a temperature of 50°C and a pressure of 0.1 bar).

[0211] The formulations of the present invention may comprise between about 1% by weight and about 99% by weight, such as about 10% by weight (e.g., about 20% by weight, for example about 50% by weight) to about 90% by weight of coated particles, with the remainder consisting of a carrier system and / or other pharmaceutically acceptable excipients.

[0212] The formulations of the present invention can be in the form of liquids, sols, or gels, and can be applied via surgical application devices, such as needles or catheters, to form reservoir formulations.

[0213] In any case, the preparation of suitable formulations can be carried out non-inventively by a person skilled in the art using conventional techniques. Therefore, the formulations of the present invention and dosage forms comprising them can be formulated together with conventional pharmaceutical additives and / or excipients used in the art for preparing pharmaceutical formulations, and then incorporated into various pharmaceutical preparations and / or dosage forms using standard techniques (see, for example, Lachman et al., 'The Theory and Practice of Industrial Pharmacy', Lea and Febbiger, 3rd ed. (1986); 'Remington: The Science and Practice of Pharmacy', Troy (ed.), University of the Sciences in Philadelphia, 21st ed. (2006); and / or 'Aulton's Pharmaceutics: The Design and Manufacture'). References to the document mentioned in the 'History of Medicines', Aulton and Taylor (eds.), Elsevier, 4th edition, 2013, and all relevant public information in all such documents are hereby incorporated by reference.

[0214] According to another aspect of the invention, a method for preparing the formulations of the invention is provided, the method comprising mixing coated particles as described herein with, for example, an aqueous carrier system as described herein.

[0215] For intramuscular or, especially, subcutaneous injection, the formulations of the present invention can be in the form of sterile injectable dosage forms, which can be administered via surgical application devices (e.g., syringes with injection needles, catheters, etc.) to form reservoir formulations.

[0216] Particularly preferred administration devices include autoinjectors, which are medical devices known to those skilled in the art specifically designed for patients to administer predetermined doses of medication themselves, typically single-use spring-loaded syringes, and include any of those administration devices currently known in the art for GLP1RA administration.

[0217] Therefore, an injectable dosage form comprising the formulation of the present invention is further provided, wherein the formulation is contained in a reservoir connected to and / or associated with an injection device (e.g., a syringe with an injection needle).

[0218] Alternatively, the formulations of the present invention may be stored before being loaded into a suitable injectable delivery device (e.g., a syringe with an injection needle), or may be prepared on the spot even before being loaded into such a delivery device.

[0219] Therefore, sterile injectable dosage forms may comprise containers or reservoirs in communication with an injection device. The formulations of the present invention may be pre-loaded into said containers or reservoirs, or may be loaded at a point before use, or may comprise one or more reservoirs in which the coated particles and aqueous carrier systems of the formulations of the present invention are separately contained and mixed before and / or during injection.

[0220] Therefore, a multi-part kit is further provided, comprising:

[0221] (a) Coated particles of the formulation of the present invention; and

[0222] (b) The carrier system of the formulation of the present invention.

[0223] And a multi-part kit containing coated particles of the formulations of the present invention, and instructions for end users to mix these particles with the carrier system according to the present invention.

[0224] Further, a pre-loaded injectable dosage form as described above is provided, but modified, comprising at least two chambers, one chamber containing coated particles of the formulation of the present invention, and the other chamber containing an aqueous carrier system of the formulation of the present invention, wherein mixing occurs before and / or during injection to form a suspension or other substance.

[0225] The formulations of this invention can be used in human medicine. Specifically, the formulations of this invention can be used for any indication for which GLP1RA is approved or otherwise known to be applicable. In particular, the formulations of this invention can be used to treat metabolic conditions or disorders.

[0226] The term 'metabolic symptoms or conditions' will be understood to include endocrine disorders and / or conditions, and therefore include conditions of glands of the endocrine system (including the adrenal glands, thyroid gland, pituitary gland, and specifically the pancreas), and therefore include all types of diabetes, such as hyperglycemia, type 1 diabetes, type 2 diabetes, impaired glucose tolerance, non-insulin-dependent diabetes, juvenile-onset diabetes, and gestational diabetes; symptoms related to weight and / or overeating, such as obesity, which may be induced by habit or exogenous sources (such as steroids and antipsychotic drugs), as well as binge eating and bulimia nervosa. The term may also include the medical term 'metabolic syndrome'.

[0227] The body mass index (BMI; calculated as a person’s weight (kg) divided by the square of the subject’s height (m)) of a human subject with obesity (obese subject) may be at least about 25, such as at least about 27, including at least about 30, such as in the range of about 30 to about 40 (e.g., about 35), or even greater than 40.

[0228] Preferred metabolic conditions include type 2 diabetes and obesity, which, as mentioned above, are often considered comorbidities. In addition to diabetes (such as type 2 diabetes), overweight-related comorbidities that can also be treated with the formulations of this invention include hypertension, dyslipidemia, high cholesterol, and obstructive sleep apnea.

[0229] The formulations of the present invention are applicable to therapeutic, palliative and / or diagnostic treatment of any of the above-mentioned metabolic disorders or symptoms, as well as to preventive treatment (including prevention and / or elimination of the worsening and / or exacerbation of symptoms).

[0230] This can include delaying or preventing the progression of diabetes, such as type 2 diabetes, delaying the progression of impaired glucose tolerance (IGT) to insulin-dependent type 2 diabetes, and / or delaying the progression of insulin-independent type 2 diabetes to insulin-dependent type 2 diabetes. It can also include reducing food intake, reducing weight, suppressing appetite, inducing satiety, reducing gastric motility, and / or delaying gastric emptying.

[0231] The formulations of the present invention may also be used to treat and / or prevent (as described above, which may include eliminating exacerbations and / or aggravations) conditions, symptoms, or diseases that may be caused by metabolic conditions or disorders, including hypertension and more serious adverse cardiovascular events such as myocardial infarction, stroke, angina pectoris, heart failure, and other cardiovascular conditions; non-alcoholic fatty liver diseases such as non-alcoholic fatty liver and non-alcoholic steatohepatitis; and other neurodegenerative conditions such as Alzheimer's disease and Parkinson's disease, as well as other conditions including nerve damage, skin conditions such as chronic diabetic ulcers, diabetic ketoacidosis, eye conditions such as retinopathy and / or kidney problems (such as chronic renal failure), such as kidney disease, gum disease and / or oral problems, cancer, and / or sexual dysfunction.

[0232] In the treatment of any of the above-mentioned conditions, GLP1RA may be combined with other treatments known to be used to treat the relevant conditions.

[0233] This includes type 2 diabetes receptor analogs, such as GIP, glucagon, sodium-glucose cotransporter, amylin, and gastrin / YY peptide, as well as known diabetes treatments such as insulin, biguanides (e.g., buformin, phenformin, specifically 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 postprandial glucose regulators (e.g., reserpine). Repaglinide and nateglinide, peroxisome proliferator-activated receptor γ (PPAR) or thiazolidinediones (such as rosiglitazone, troglitazone, or pioglitazone), dipeptidyl peptidase-4 (DPP-4) inhibitors (gliptin, such as vildagliptin, sitagliptin, saxagliptin) (axagliptin), linagliptin, septagliptin, teneligliptin, gemigliptin, and alogliptin) and / or SGLT2 inhibitors (such as dapagliflozin, canagliflozin, empagliflozin, and remogliflozin).

[0234] It also includes known obesity treatments such as orlistat, cetilistat, lorcaserin, sibutramine, rimonabant, metformin, berberine, forskolin, and combination drugs such as amylin / pramlinatide, phentermine / topiramate, naltrexone / bupropion, superabsorbent hydrogels (e.g., Gelesis100), and / or statins.

[0235] The formulations of the present invention (with or without the aforementioned combination therapies) may also be used in conjunction with blood glucose meters and continuous glucose monitors, and / or in conjunction with more invasive techniques such as islet cell transplantation or bariatric surgery.

[0236] Additionally, as described below, we have found that injection of the formulations of the present invention may cause a mild inflammatory reaction. Such reactions can be alleviated by co-administration with an injectable anti-inflammatory agent.

[0237] Appropriate anti-inflammatory agents that can be used in this regard include butylpyrazolidines (such as phenylbutazone, mofebutazone, oxyphenbutazone, clofezone, kebuzone, and suxibuzone); acetic acid derivatives and related substances (indomethacin, sulindac, tolmetin, zomepirac, diclofenac, alclofenac, bumadizone, etodolac, lonazolac, fentiazac, acemetacin, difenpiramide, and oxametacin). Proglumetacin, ketorolac, aceclofenac, and bufexamac; oxicam (such as piroxicam, tenoxicam, droxicam, lornoxicam, and meloxicam); propionic acid derivatives (such as ibuprofen, naproxen, ketoprofen, fenoprofen, fenbufen, benoxaprofen, supraprofen, pirprofen, flurbiprofen, indoprofen, and tiaprofenic acid). (Acids), oxaprozin, ibuprofen, dexibuprofen, flunoxaprofen, aminoprofen, dexketoprofen, vedaprofen, carprofen, and tepoxalin; fenamates (such as mefenamic acid, tolfenamic acid, and flufenamic acid).Meclofenamic acid and flunixin), coxib derivatives (such as celecoxib, rofecoxib, valdecoxib, parecoxib, etoricoxib, lumiracoxib, firocoxib, robenacoxib, mavacoxib, and cimicoxib); other nonsteroidal anti-inflammatory drugs (such as nabumetone, niflumic acid, etc.). acid), azapropazone, glucosamine, benzyldamine, glucosaminoglycan polysulfate, proquazone, orgotein, nimesulide, feprazone, diacerein, morniflumate, tenidap, oxaceprol, chondroitin sulfate, pentosan. (polysulfate) and aminopropionitrile; corticosteroids (such as 11-dehydrocorticosterone, 11-deoxycorticosterone, 11-deoxycortisol, 11-ketoprogesterone, 11β-hydroxypregnenolone, 11β-hydroxyprogesterone) esterone), 11β,17α,21-trihydroxypregnenolone, 17α,21-dihydroxypregnenolone, 17α-hydroxypregnenolone, 17α-hydroxyprogesterone, 18-hydroxy-11-deoxycorticosterone18-hydroxycorticosterone, 18-hydroxyprogesterone, 21-deoxycortisol, 21-hydroxycortisone, 21-hydroxypregnenolone (prebediolone), aldosterone, corticosterone (17-deoxycortisol), cortisol (hydrocortisone), cortisone, pregnenolone, progesterone, flugestone (flurogestone), fluorometholone, medrysone (hydroxymethylprogesterone), prebediolone Acetate (21-acetoxypregnenolone), chloroprednisone, cloprednol, difluprednate, fludrocortisone, fluocinolone, fluperolone, fluprednisolone, loteprednol, methylprednisolone, prednicarbate, prednisolone Prednisone, tixocortol, triamcinolone, alclometasone, beclometasone, betamethasone, clobetasol, clobetasone, clocortolone, desoximetasone, dexamethasone, diflorasone, difluocortolone, flucloroloneFlumetasone, fluocortin, fluocortolone, fluprednidene, fluticasone, fluticasone furoate, halometasone, meprednisone, mometasone, mometasone furoate furoate, paramethasone, prednylidene, rimexolone, ulobetasol (halobetasol), amcinonide, budesonide, ciclesonide, deflazacort, desonide, formocortal, fluclorolone acetonide, flucloronide, fluroxycortide, flurandrenolone, flurandrenolide, flunisolide, fluocinolone acetonide, fluocinonide, halcinonide, and triamcinolone Acetone; quinoline derivatives (such as oxycinchophen); gold preparations (such as sodium aurothiomalate, sodium aurothiosulfate, auranoofin, aurothioglucose, and aurothioprol); penicillamine and similar agents (such as buccilamine); and antihistamines (such as akrivastin, alimethazine, antazolin, astemizol, azatadin, azelastin, bamipin, bilastin, bromdifenhydramin, bromfeniramin, buklizine, and cetirizine).Cinnarizine, Cyklizine, Cyproheptadine, Deptropine, Desloratadine, Dexbromfeniramin, Dexklorfeniramin, Difenylpyralin, Dimenhydrinat, Dimetinden, Doxylamin, Ebastin, Epinastine n), fenindamin, feniramin, fexofenadin, histapyrrodin, hydroxietylprometazin, isotipendyl, karbinoxamin, ketotifen, kifenadin, klemastin, klorcyklizin, chlorpheniramine, carfexobin, carfexobin min), kloropyramin, levocetirizine, loratadine, mebhydrolin, mekitazin, meklozin, mepyramin, metapyrilen, metdilazin, mizolastin, oxatomide, oxomemazine, pimexicillin, prometaxine The following are listed as anti-inflammatory agents: pyrrobutamin, rupatadin, sekifenadin, talastin, tenalidin, terfenadine, tiazinam, tietylperazin, tonzylamin, trimetobenzamid, tripelennamin, triprolidine, and tritokvalin. Any one or more combinations of the above anti-inflammatory agents can be used.

[0238] Preferred anti-inflammatory agents include nonsteroidal anti-inflammatory drugs (NSAIDs) such as diclofenac, ketoprofen, meloxicam, aceclofenac, flurbiprofen, parecoxib, ketoralac tromethamine, or indomethacin.

[0239] Subjects may receive (or may have received) one or more of the above-described co-therapeutic agents and / or anti-inflammatory agents separate from the formulations of the present invention, meaning that they receive one or more of those other therapeutic agents in prescribed doses before, in addition to and / or after treatment with the formulations of the present invention.

[0240] When GLP1RA / its salts are “combined” with other such therapeutic agents, the active ingredients can be administered together in the same formulation or individually (simultaneously or sequentially) in different formulations (hereinafter referred to as “combination products”).

[0241] Such combination products provide for the combined administration of GLP1RA with other therapeutic agents, and can therefore be presented as individual formulations (where at least one of those formulations is a formulation of the present invention, and at least one of the individual formulations contains other therapeutic agents), or as combination formulations (i.e., formulations) (i.e., as a single formulation comprising GLP1RA / salt and other therapeutic agents).

[0242] In this regard, another therapeutic agent may be co-present with GLP1RA in one or more cores forming part of the formulation of the present invention as described above, or may be formulated using the same or similar GLP1RA coating process as described above, which may allow the release of other therapeutic agents on the same or different timescales.

[0243] Therefore, a pharmaceutical formulation of the present invention is further provided, the pharmaceutical formulation further comprising a therapeutic agent for treating metabolic conditions such as type 2 diabetes or obesity as defined above, and / or an anti-inflammatory agent.

[0244] The pharmaceutical formulations of the present invention that may be mentioned include those that do not contain and / or are substantially free of said anti-inflammatory agents. “Substantially free of anti-inflammatory agents” means 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 about 0.01% by weight) of one or more of the aforementioned anti-inflammatory agents.

[0245] In such formulations of the present invention, additional therapeutic agents may be included in the following manner:

[0246] (1) Formulated together with GLP1RA within the core of the formulation of the present invention (the formulation is referred to below as a 'combination core formulation'); or

[0247] (2) Dissolve and / or suspend it in an aqueous carrier system of the formulation of the present invention (the formulation is referred to below as a 'combination formulation').

[0248] In the above embodiment (2), other therapeutic agents may be present in the formulation of the present invention in any form, wherein they are separate from the core containing GLP1RA. This can be achieved, for example, by directly dissolving or suspending the active ingredient in an aqueous medium of the formulation of the present invention, or by presenting the active ingredient in a form in which its release after injection is controlled, similar to that of GLP1RA.

[0249] The latter option can be achieved, for example, by providing additional therapeutic agents in the form of additional particles suspended in an aqueous carrier system of the formulation of the present invention, the additional particles having an average diameter based on weight, number, or volume between about 10 nm and about 700 μm, and comprising a core containing a therapeutic agent and / or anti-inflammatory agent for treating metabolic disorders as defined above, the core being at least partially coated with one or more coating materials as described above (the formulation is referred to below as a 'combined suspension').

[0250] A further provision of the present invention is a pharmaceutical formulation, said pharmaceutical formulation being in the form of a multi-part kit comprising the following components:

[0251] (A) The pharmaceutical formulation of the present invention; and

[0252] (B) Pharmaceutical formulations containing therapeutic agents and / or anti-inflammatory agents that can be used to treat metabolic disorders as defined above.

[0253] Components (A) and (B) are each provided in a form suitable for combined application with another component.

[0254] Although component (B) of the multi-part kit presented above may differ from component (A) (i.e., the formulation of the present invention) in terms of its chemical composition and / or physical form, it may also be in a form substantially the same as or at least similar to the formulation of the present invention containing GLP1RA, i.e., in the form of multiple particles suspended in a (e.g., aqueous) carrier system, said particles:

[0255] (a) Having an average diameter based on weight, number, or volume between about 10 nm and about 700 μm; and

[0256] (b) Containing a solid core, the solid core containing the other therapeutic agent, the core being at least partially coated with one or more (e.g., inorganic) materials.

[0257] Furthermore, although the coated cores containing another therapeutic agent may differ in their chemical composition and / or physical form in such preferred multipart kits and the combined suspensions presented in Example (2) above, it is preferred that the inorganic coating material used is the same as or similar to the coating used in the formulations of the present invention containing GLP1RA. This means that the other therapeutic agent is coated by one or more inorganic coatings as described above, such as one or more metal- or metal-like compounds, such as metal oxides or metal-like oxides, such as iron oxide, titanium dioxide, zinc sulfide, more preferably zinc oxide, silicon dioxide and / or aluminum oxide, wherein the coating material may (on a single or combined basis) consist substantially of such oxides (e.g., greater than about 80%, such as greater than about 90%, such as about 95%, such as about 98%), and more specifically, the inorganic coating comprises a mixture of the following substances:

[0258] (i) Zinc oxide; and

[0259] (ii) one or more other metal oxides and / or metal-like oxides.

[0260] The atomic ratio ((i):(ii)) is at least about 1:6 and at most and includes about 6:1.

[0261] Preferably, the atomic ratio ((i):(ii)) is at least about 1:1 and at most and includes about 6:1.

[0262] In any event, and to avoid doubt, all aspects of the formulations of the present invention containing GLP1RA disclosed and / or claimed herein, including preferred aspects, are equally applicable to aspects and / or preferences comprising one or more of the additional therapeutic agents described above. For the avoidance of doubt, these aspects, preferences, and features, individually or in combination, are incorporated herein by reference.

[0263] Therefore, all combination products, including the aforementioned combination core formulations, combination suspensions, and multipart kits, are intended for human medical use, and specifically, GLP1Ra is approved for or is known to be applicable to any of the indications described above, such as metabolic disorders or conditions as defined above, such as type 2 diabetes and / or obesity.

[0264] In some cases, such additional therapeutic agents, including some that can be used to treat, for example, metabolic disorders or conditions as defined above (such as type 2 diabetes and / or obesity), may be referred to as “standard care” in relation to a specific condition. Technicians will understand that the term “standard care” includes the treatment procedures that clinicians should follow and / or expect to follow when treating certain types of patients, diseases, and / or clinical conditions. In some new or less understood conditions, standard care may change and / or evolve over time.

[0265] According to another aspect of the invention, a method for preparing a multi-component kit as defined above is provided, the method comprising combining a component (A) as defined above with a component (B) as defined above, such that the two components are suitable for combined administration with each other.

[0266] Combining the two components with each other includes, in a multi-part kit, components (A) and (B) can:

[0267] (i) provided as separate formulations (i.e., independent of each other), and subsequently assembled for combined use in combination therapy; or

[0268] (ii) Packaged and presented together as individual components of a 'combination pack' for use in combination therapy.

[0269] Therefore, a multi-part kit as defined above is further provided, wherein components (A) and (B) are packaged and presented together as separate components of a combination package for combined use in combination therapy, and a multi-part kit comprising:

[0270] (I) one of components (A) and (B) as defined herein; and

[0271] (II) Instructions for using the component in combination with another component of the two components.

[0272] As described above, the multipart kits described herein may contain more than one formulation comprising an appropriate amount / dosage of GLP1RA / salt, and / or more than one formulation comprising an appropriate amount / dosage of other therapeutic agents, for repeated administration as described above.

[0273] In this regard, regarding the multi-component kit described herein, 'combined administration' includes the sequential, separate and / or simultaneous administration of components (A) and (B) of the kit during the treatment of the relevant symptoms.

[0274] Therefore, the term 'combination' includes the administration of one or more of two formulations before, after, and / or at the same time (optionally repeated). When used in this context, the terms 'simultaneous administration' and 'administered at the same time' include the administration of a single dose of GLP1RA / salt and other therapeutic agents at intervals of 48 hours (e.g., 24 hours) from each other.

[0275] With respect to any of the above-described combination products according to the invention, the corresponding formulation is administered in such a manner (or, in the case of a multi-component kit, the two components are optionally administered repeatedly or in combination with each other) that, compared to administering alone (e.g., repeatedly, as described herein) a formulation containing GLP1RA / salt (e.g., the formulation of the invention) during the course of treatment of the condition in the absence of other components, the manner described above may provide a greater beneficial effect to the subject during the same course of treatment.

[0276] Determining whether a combination of products provides greater therapeutic benefit and during treatment will depend on the condition being treated and / or its severity, but can be routinely accomplished by a technician.

[0277] For example, a physician may initially administer a formulation containing only the ingredients of this invention 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 by any other components of the formulation).

[0278] The doctor may then administer one or more of the following as described above:

[0279] • Component (B) of the multi-part kit as described above,

[0280] • Combined core formulation,

[0281] Combination formulations, and / or

[0282] Combined suspension

[0283] Each of these contains an anti-inflammatory agent as described above.

[0284] Other active ingredients / therapeutic agents described above that may be used in the combination products according to the invention may be provided in the form of (e.g., pharmaceutically acceptable) salts, including any such salts known in the art and described in medical literature (such as Martindale – The Complete Drug Reference, 38th edition, Pharmaceutical Press, London (2014) and references therein) (the relevant disclosures in all such literature are hereby incorporated by reference).

[0285] The amount of other active ingredients / therapeutic agents that may be used in the combination products according to the present invention must be sufficient to exert their pharmacological effects.

[0286] Therefore, the dosage of such other active ingredients that can be administered to a patient should be sufficient to influence the therapeutic response within a reasonable and / or relevant timeframe. Those skilled in the art will recognize that the precise dosage and choice of composition, as well as the most appropriate delivery regimen, will be influenced not only by the nature of the other active ingredients, but also, in particular, by the pharmacological properties of the formulation, the route of administration, the nature and severity of the condition being treated, the recipient's physical and mental condition, the patient's age, condition, weight, sex, and response, as well as the stage / severity of the disease and genetic differences between patients.

[0287] Since the application of the formulations of the present invention can be continuous or intermittent (e.g., by bolus injection), the dosage of such other active ingredients can also be determined by the timing and frequency of application.

[0288] In any case, a licensed physician or other technical person will be able to routinely determine the actual dose of any particular additional active ingredient most suitable for an individual patient, and the doses of the aforementioned additional active ingredients include those known in the art and described in medical literature for the drug in question, such as Martindale: The Complete Drug Recipe, 38th edition, Medical Publishers, London (2014) and references therein, all of which contain relevant disclosures which are hereby incorporated by reference.

[0289] The use of the formulations of the present invention can reduce any burst-release effect as defined above (e.g., the maximum concentration shortly after administration) and / or by reducing the C0 in the plasma concentration-time curve. max To control the dissolution rate of GLP1RA and its impact on pharmacokinetic curves.

[0290] The formulations of the present invention can also provide release and / or pharmacokinetic profiles that increase the length of GLP1RA release from the formulation.

[0291] These factors not only reduce the frequency or time required to administer the compound to people with related metabolic symptoms or conditions, but also allow patients more time as outpatients, resulting in a better quality of life, and offer the advantage of fewer / lower frequency of inconvenient and / or painful injections.

[0292] The formulations of the present invention also have the advantage of providing a lower daily exposure to the relevant active ingredients by controlling the release of the active ingredients at a stable rate over an extended period of time, which is expected to reduce unwanted side effects.

[0293] The formulations and methods described herein may also have the following advantages: in the treatment of related metabolic symptoms, compared with any similar treatment known in the prior art, the formulations and methods described herein may be more convenient, more effective, less toxic, have a wider range of activities, be more potent, produce fewer side effects, or have other useful pharmacological properties.

[0294] Whenever the word 'about' is used herein, such as in the context of quantities (e.g., number, concentration, size (size and / or weight), dosage, time period, pharmacokinetic parameters, etc.), relative quantities (percentage, weight ratio, size ratio, atomic ratio, aspect ratio, proportion, coefficient, fraction, etc.), relative humidity, lux, temperature, or pressure, it should be understood that these variables are approximate and therefore may differ from the figures specified herein by ±15%, such as ±10%, for example ±5%, and preferably ±2% (e.g., ±1%). This is also true even if such figures are first presented as percentages (e.g., 'about 15%' could mean ±15% of the number 10, i.e., any value between 8.5% and 11.5%).

[0295] The present invention is illustrated by the following examples with reference to the accompanying drawings, but is in no way limited thereto, wherein Figure 1 The results of coating integrity assays for two batches of coated liraglutide microparticles are shown. Figure 2 The reduction in drug load was shown in coated and uncoated particles after γ-irradiation. Figure 3 The semi-logarithmic plot of mean plasma concentration (ng / mL) versus time (day) after a single subcutaneous administration of liraglutide to rats at different formulations and dosage levels is shown. Figure 4 The average plasma concentrations over the first 72 hours in the same study are shown; Figure 5 The in vitro release results of two batches of coated liraglutide microparticles are shown; Figure 6 The figure shows a semi-logarithmic plot of mean plasma concentration (ng / mL) versus time (day) after a single subcutaneous administration of liraglutide to miniature pigs with different formulations. Figure 7The in vitro release results of four batches of coated liraglutide microparticles are shown; and Figure 8 and Figure 9 The mean plasma concentration (ng / mL) of liraglutide in rats after a single subcutaneous administration of different formulations versus time (days) is shown. Figure 8 7 days Figure 9 A semi-logarithmic graph (42 days).

[0296] Example

[0297] Example 1

[0298] Coated liraglutide microparticles

[0299] R&D grade liraglutide (MedChemExpress, New Jersey, USA) 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).

[0300] The particle size distribution determined by laser diffraction (SALD-7500nano (Shimadzu Corporation of Japan), 405nm laser) is as follows: %D(10): 2.0μm, %D(50): 7.3μm, %D(90): 23.8μm.

[0301] The raw materials are dried in a small spray dryer (B-290, with a dehumidifier B-296 featuring dual-fluid nozzles; Germany) Labortechnik GmbH Labortechnik GmbH, Germany) spray-drying is performed by dispersing the raw materials in purified water (0.8–2 MΩ / cm). 2 The mixture forms a milky white liquid, wherein the liraglutide concentration is 15 wt%.

[0302] The dispersion was spray-dried at an inlet temperature of 115°C and a suction rate of 100% (approximately 35m). 3 The pump rate was 8% (3.8 mL / min), nozzle cleaning was 2, volumetric flow rate was 35 mm (N2, approximately 600 L / h), and outlet temperature was 73°C. The yield was approximately 77.5%.

[0303] The spray-dried material was analyzed using a Nexera UPLC-UV-DAD (Shimadzu Corporation, Japan) with a SunShell nC18-WP, 4.6×100mm, 2.6μm particle size column (Chromanik Technologies Inc., Japan). The results showed that the liraglutide content in the spray-dried material was 100.1±0.2% (relative amount).

[0304] The particle size of the spray-dried material was determined as follows: %D(10): 1.7 μm, %D(50): 8.2 μm, %D(90): 21.2 μm.

[0305] The spray-dried liraglutide particles were first coated with three layers of pure alumina using the following method:

[0306] a. Close the valve on the pipeline between the pump and the ALD reactor.

[0307] b. Then open the valve on the trimethylaluminum (TMA) precursor bottle for 1 second to allow the evaporated metal-containing precursor to fill the ALD reactor for 1 second.

[0308] c. Close the valve of the precursor bottle and allow the chamber to stand for 30 seconds (soaking time) before opening it to the pump again to ensure that the metal-containing precursor vapor reacts with the surface of the drug particles.

[0309] d. Thereafter, the ALD reactor is pumped for 9 seconds.

[0310] e. Repeat step ad above 20 times.

[0311] The chamber is then purged with a continuous stream of nitrogen to remove unreacted reagents and organic gases. Thereafter, steps (a) through (e) are essentially repeated, except that water is used as a second reagent to form a discrete alumina layer on the surface of the active ingredient particles. Subsequent purging pulses of nitrogen are then provided in a continuous stream to remove gaseous water and organic gases.

[0312] Repeat the above procedure twice to form a total of three initial aluminum oxide atomic layers.

[0313] Subsequently, in step (b) above, using diethylzinc (DEZ) and then water as precursors, three zinc oxide layers are applied by repeating the ALD step above to form a total of three zinc oxide layers. Then, an alumina layer is applied using the same precursor mentioned in section (b) above. This provides a total of four atomic layers of mixed oxide layers with an atomic ratio of 1:3 (Al₂O₃:ZnO).

[0314] This is repeated a total of 9 times to form a mixed oxide (1:3Al2O3:ZnO) layer with a total of 40 atomic layers (30 ZnO layers and 10 Al2O3 layers).

[0315] The powder is then removed from the ALD reactor and deagglomerated using an acoustic sieve (Tsutsui Sonic Agitated Sifting Machine SW-20AT) with a sieve having a 20μm mesh size.

[0316] The resulting deagglomerated powder was resampled into the ALD reactor and coated with a mixed oxide (1:3 Al2O3:ZnO) layer having a total of 40 atomic layers, following the same procedure. The deagglomeration and coating steps were repeated twice to produce samples with a pure alumina underlayer and four discrete mixed oxide coatings (1:3 Al2O3:ZnO atomic ratio).

[0317] The resulting particles were then divided into two batches. For the first batch, an additional pure alumina outer layer was applied using the same pure alumina coating procedure as described above.

[0318] For the second batch, the procedure described above for preparing 40 mixed Al2O3:ZnO(1:3) oxide layers was repeated twice under deagglomeration conditions to provide two additional discrete mixed oxide coatings, followed by post-treatment with an additional pure alumina outer layer as described above.

[0319] Therefore, the first batch contains 3 Al2O3 (inner) layers, 4 x 40 1:3 Al2O3:ZnO mixed oxide layers, and 3 Al2O3 (outer) layers, and the second batch contains 3 Al2O3 (inner) layers, 6 x 40 1:3 Al2O3:ZnO mixed oxide layers, and 3 Al2O3 (outer) layers.

[0320] The particle size of a batch was determined by suspending the coated particles in a solution of 0.1% Span 85 (Sigma-Aldrich, Missouri, USA) in heptane (Merck, Germany). The particle size distribution, as measured by laser diffraction, is as follows:

[0321] Batch 1 %D(10): 3.5±0.3μm; %D(50): 10.7±0.6μm; %D(90): 23.6±0.6μm;

[0322] Batch 2 %D(10): 3.2±0.2μm, %D(50): 9.7±0.6μm, %D(90): 22.3±0.8μm.

[0323] The drug loading of the coated batch was determined as follows: Samples were etched to dissolve the coating in a solution of 43% acetonitrile (Spectrascan, Scotland), 45% water (PanReac, Spain), and 12% (v / v) phosphoric acid (Merck, Germany), then diluted to 160 μg / mL with 10% aqueous acetonitrile + 0.1% trifluoroacetic acid (Merck, Germany) (mobile phase A) and injected into a Kinetex C18 PS column (150 × 4.6 mm, 2.6 μm particle size, Phenomenex, California, USA) for quantification. The HPLC system (Prominence-IHPLC-UV-DAD, Shimadzu Corporation, Japan) of Bachem AG Ltd., CA, USA was used; mobile phase B: 80% acetonitrile aqueous solution + 0.1% trifluoroacetic acid; injection volume 10 μL, autosampler temperature 15°C, oven temperature 25°C; gradient elution was used at a flow rate of 1.0 mL / min; UV absorbance was measured at 220 nm; single-point calibration was used for determination; the retention time of liraglutide was 6.8 min; standard substance: 99.4% total liraglutide (as determined by assay), 99.1% purity (as determined by HPLC, Bachem AG, Switzerland).

[0324] The drug loading of the first batch was 61.9 ± 0.2%, and that of the second batch was 53.1 ± 0.4%. In both cases, the purity of the coated liraglutide was measured to be 98.9%. Therefore, no significant decrease in the purity of the coated liraglutide was observed, indicating that it is resistant to the ALD process and does not undergo degradation.

[0325] To determine the integrity of the coating, two batches of samples were suspended in dimethyl sulfoxide (Rathburn, UK) at a concentration of 0.4 mg liraglutide per mL of solvent and stirred on a top-mounted stirrer for up to 72 hours. Intermittent samples were removed, centrifuged at 6000 rpm (EBA 20, Hettich, Germany) for 7 min, and the supernatant was diluted in mobile phase A (as above) before being injected into the HPLC system described above for quantification. Results are shown in... Figure 1 (Batch 1 (circle), Batch 2 (square)) indicates that defects in the coating are minimal, therefore any burst release is expected to be low.

[0326] Sample materials from batch 1 and from uncoated spray-dried material were subjected to doses ranging from 20.4–20.6 kGy and 23.5–25.3 kGy, respectively. 60 Co source (Ionisos Baltics, Alicoo, Estonia) Company (Ionisos Baltics) Alliku, Estonia) underwent gamma irradiation at sterilization levels. The drug load of the coated material was determined using the same method described above and compared to the drug load of the uncoated material. At approximately 21 kGy of gamma irradiation, the absolute reduction in drug load of the first batch of material was less than 5% unit, indicating that this material could be terminally sterilized using this method in a sufficiently non-destructive manner. Results are shown in… Figure 2 The error bars show the standard deviation (n=3), and ns indicates no significant difference (p>0.05), as shown by using Dunn– (As determined by corrected two-way ANOVA). The purity of liraglutide in the treated material was determined using the same method as described above, and was above 95% in all samples, confirming the results of the drug loading assay.

[0327] Example 2

[0328] Formulation I of the invention

[0329] The coated liraglutide particles (from batches 1 and 2 of Example 1 above) were suspended in a suitable aqueous carrier, such as phosphate-buffered saline at a concentration between 5 mM and 30 mM and 100 mM tromethorphan buffer, and the volume was made up with water for injection to provide a suspension with a concentration of approximately 100 mg liraglutide per milliliter of aqueous carrier. The pH of the resulting formulation was measured to be approximately 7.4.

[0330] Example 3

[0331] Formulation II of the invention

[0332] Suspensions of two coated liraglutide microparticles (batch 1 and batch 2, prepared according to the method described in Example 1 above) were prepared by adding an appropriate volume of veterinary hyonate (1 mM phosphate buffer containing 1 wt% hyaluronic acid solution, pH 7.4) to the test vials to achieve a liraglutide concentration corresponding to 10 mg / mL. The vials were then tapped at least 10 times to remove any material that might have precipitated at the bottom of the test vials. The mixture was then vortexed for approximately 60 seconds to ensure homogeneity of the suspension.

[0333] Example 4

[0334] In vivo study

[0335] On the day of administration, sixteen male Sprague Dawley rats weighing approximately 300g were provided by Charles River Labs (UK). The animals were randomly divided into groups of four.

[0336] Before injection, the hair in the intended application area was clipped, and the injection site was marked. A single subcutaneous injection (approximately 0.08 mL for batches 1 and 2, group 1 and 3 respectively, as indicated in Table 1 below, and 0.17 mL for batch 2, group 2, as indicated in Table 1 below) was administered subcutaneously to the flank of each rat using a 23G needle. A control liraglutide solution (Victoza, Novo Nordisk, Denmark) was diluted to 0.6 mg / mL in saline solution for injection (group 4, as indicated in Table 1 below), drawn into a 1 mL syringe, and administered subcutaneously to the flank of each rat using a 23G needle. Administration was performed no more than 30 minutes after preparation.

[0337] After administration, the administration site was marked on each animal and kept hair-free at the site during the study. The administration site was observed at 1 hour and 24 hours post-administration, and then daily until the last sampling point.

[0338] Table 1

[0339]

[0340]

[0341] For groups 1 through 3, blood samples (approximately 0.2 mL) were collected from the jugular vein into K2EDTA (dipotassium ethylenediaminetetraacetate) tubes at the following time points: 1 hour, 3 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 120 hours, 168 hours, 251 hours, 384 hours, 480 hours, 576 hours, and 672 hours after administration. For group 4, samples were collected at the following time points: 1 hour, 2 hours, 3 hours, 6 hours, 9 hours, 12 hours, 24 hours, and 48 hours after subcutaneous administration. The actual sampling time was recorded. Plasma was separated as soon as possible after blood sampling by centrifugation (1500 g, 4°C, for 10 minutes) and stored at -80°C until analysis.

[0342] After the study was completed, all plasma samples were deep-frozen on dry ice before being shipped for analysis. Animals were euthanized on the last day of the study.

[0343] Plasma concentrations of liraglutide were determined using LC-MS / MS. Study samples were prepared using an EVO-2 liquid handling robot (Tecan, Austria) by pipetting 35 μL of rat plasma into 96-well plates and adding 75 μL of internal standard working solution. The 96-well plates were then shaken for 15 minutes and centrifuged. All samples were then injected into a UPLC-MS / MS system (Waters, MA, USA). An ACQUITY UPLC Protein BEH C4 column was used. Separation was performed using a 1.7 μm, 2.1 mm x 50 mm (Waters Corporation, Massachusetts, USA) mobile phase A and acetonitrile as mobile phase B at 60 °C with a 0.3% formic acid aqueous solution.

[0344] Non-compartmental analysis (NCA) was performed using Phoenix WinNonlin version 8.3 software (Certara, USA). The pharmacokinetic (PK) analysis of liraglutide in plasma was evaluated. Cp was derived from the observed plasma concentration data. max and t max AUC is assessed by integrating the plasma concentration versus time curve, using linear interpolation for increasing plasma levels and logarithmic interpolation (linear for the ascending segment and logarithmic for the descending segment) for decreasing plasma levels. For AUC... ∞ Calculations were made up to the last point (AUC) showing the measurable plasma concentration. last The area of ​​the curve is calculated, and then extrapolated to infinity using the concentration in the last quantifiable sample and the first-order rate constant λz associated with the end of the curve. 1 / 2,z From ln2 / λ z calculate.

[0345] result

[0346] Figure 3 The dose-normalized plasma concentrations of liraglutide over four weeks following a single subcutaneous administration of various formulations are presented. Figure 4 The same plasma concentration curves are shown for the first 72 hours.

[0347] Plasma pharmacokinetic parameters are also presented in Table 2 below as mean values ​​(standard deviations are provided in parentheses) for the six rat groups, where:

[0348] ·'t max 'This is the time to reach peak concentration, expressed in hours.'

[0349] ·'C max ' is the highest concentration found in the analysis, expressed in ng / mL.

[0350] ·'t last 'This is the last detectable concentration, expressed in hours.

[0351] ·'t 1 / 2,z 'This is the terminal half-life, expressed in hours'

[0352] ·'AUC ∞ ' is the area under the concentration-time curve extrapolated to infinity, expressed in ng*h / mL.

[0353] • 'F' is relative bioavailability, expressed as a percentage.

[0354] ·'C max / D' is the maximum concentration normalized to 1 mg / kg, expressed in ng / mL / mg / kg rat body weight.

[0355] ·'AUC last / D' is the area under the curve of blood concentration versus time, normalized to 1 mg / kg up to the last detectable concentration, expressed as ng*h / mL / mg / kg rat body weight.

[0356] ·'AUC ∞ / D' is the area under the concentration-time curve normalized to 1 mg / kg up to infinity, expressed as ng*h / mL / mg / kg rat body weight.

[0357] ·'Fr.Rel. 0-24h 'Is the percentage of the concentration released during the first 24 hours of the time curve, expressed as a percentage, up to infinity.

[0358] Table 2

[0359] Parameters Group 1 Group 2 Group 3 Group 4 [cat max ]] 21(6.0) 21(6.0) 18(6.9) 6(0) [C max ]]> 799(176) 670(61.2) 391(81.6) 343(49.5) <![CDATA[t last ]]> 672 672 672 48 [CAT 1 / 2,z ]]> 79.4(14.8) 153(89.0) 142(27.6) 4.43(0.71) AUC ∞ ]]> 30100(7089) 48775(5545) 26325(4892) 4620(733.8) F 47(11) 38(4.2) 41(7.6) 100(34) C max / D]] 285(62.9) 120(10.9) 140(29.2) 1710(248) AUC last / D]]> 10738(2512) 8304(1155) 9321(1798) 23090(3664) AUC ∞ / D]]> 10750(2532) 8710(990.2) 9045(1747) 23100(3669) Fr. Rel. 0-24h ]]> 42(1) 23(3) 28(2) 100

[0360] As can be seen, the plasma concentration curves of groups 1 through 3 were comparable to those of group 4 (showing complete release after only 2 days), reaching maximum plasma concentrations (C0) within the first 24 hours after administration. max The concentration then dropped rapidly until 72 hours later, and then decreased slowly but steadily over the 28-day study period.

[0361] Compared to the thicker coated formulations in Groups 2 and 3, Group 1 exhibited higher initial drug release.

[0362] Comparing groups 1 through 3, it can be seen that the systemic exposure of liraglutide is highly proportional to the administered dose, regardless of the coating thickness.

[0363] Groups 1 through 3 showed lower relative bioavailability (F) than 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.

[0364] Compared to liraglutide solution (Group 4), liraglutide (Groups 1 through 3) administered in granular form coated with a mixed oxide had a higher C content. max The levels were higher, with Group 2 releasing approximately one-fifth of the dose during Day 1, Group 3 releasing one-third, and Group 1 releasing two-fifths. For the thicker coatings (Groups 2 and 3), the differences were approximately one order of magnitude when normalized to the dose. Furthermore, the residual area (related to unreleased drug) after the last sampling time (672 hours post-dose) was <5%.

[0365] The results for Group 1 show slightly different curves, characterized by a larger dose fraction released during the first day, and C... max / D is higher and the duration is shorter.

[0366] In summary, groups 1 through 3 exhibited prolonged release curves, unlike the rapid decline observed after administration of liraglutide solution. Similarly, favorable plasma concentration-time curves were observed for all formulations of the present invention, particularly those in groups 2 and 3.

[0367] Example 5

[0368] Coated liraglutide microparticles

[0369] Coated liraglutide microparticles were prepared using a method similar to that described for Example 1, resulting in two batches corresponding to batch 3 and batch 4.

[0370] The particle size of the spray-dried liraglutide particles, as described above, was determined as follows: %D(10): 1.5 μm ± 2.5%, %D(50): 5.2 μm ± 2.9%, %D(90): 11.5 μm ± 2.3%.

[0371] Batch 3

[0372] For the third batch, the spray-dried liraglutide particles were first coated with three layers of pure alumina using the following method:

[0373] a. Close the valve on the pipeline between the pump and the ALD reactor.

[0374] b. Then open the valve on the trimethylaluminum (TMA) precursor bottle for 1 second to allow the evaporated metal-containing precursor to fill the ALD reactor for 1 second.

[0375] c. Close the valve of the precursor bottle and allow the chamber to stand for 30 seconds (soaking time) before opening it to the pump again to ensure that the metal-containing precursor vapor reacts with the surface of the drug particles.

[0376] d. Thereafter, the ALD reactor is pumped for 9 seconds.

[0377] e. Repeat step ad above 20 times.

[0378] The chamber is then purged with a continuous stream of nitrogen to remove unreacted reagents and organic gases. Thereafter, steps (a) through (e) are essentially repeated, except that water is used as a second reagent to form a discrete alumina layer on the surface of the active ingredient particles. Subsequent purging pulses of nitrogen are then provided in a continuous stream to remove gaseous water and organic gases.

[0379] Repeat the above procedure twice to form a total of three initial aluminum oxide atomic layers.

[0380] Subsequently, in step (b) above, using diethylzinc (DEZ) and then water as precursors, a zinc oxide layer is applied by repeating the ALD step above to form a total of one zinc oxide layer. This is then coated with an alumina layer using the same precursor mentioned in section (b) above. This provides a total of four atomic layers of a mixed oxide layer with an atomic ratio of 1:1 (Al₂O₃:ZnO).

[0381] This is repeated a total of 11 times to form a mixed oxide (1:1 Al2O3:ZnO) layer with a total of 25 atomic layers (11 ZnO layers and 14 Al2O3 layers), including three initial aluminum oxide atomic layers.

[0382] The powder was then removed from the ALD reactor and deagglomerated using an acoustic sieve (Tsutsui SW-20AT acoustic agitator sieve) with a sieve having a 20μm mesh size.

[0383] The resulting deagglomerated powder was re-sampled into the ALD reactor and coated with three layers of alumina, followed by a layer of mixed oxide (1:1 Al2O3:ZnO) with a total of 25 atomic layers, following the same procedure. The deagglomeration and coating steps were repeated four times to produce samples with a pure alumina underlayer and five discrete layers of mixed oxide coating (1:1 Al2O3:ZnO atomic ratio).

[0384] Therefore, the third batch contains 5 x 25 layers, which is 5 x (3 Al2O3 layers and 22 1:1 Al2O3:ZnO mixed oxide layers).

[0385] Batch 4

[0386] For batch 4, the spray-dried liraglutide particles were first coated with ten layers of pure alumina using the following method:

[0387] a. Close the valve on the pipeline between the pump and the ALD reactor.

[0388] b. Then open the valve on the trimethylaluminum (TMA) precursor bottle for 1 second to allow the evaporated metal-containing precursor to fill the ALD reactor for 1 second.

[0389] c. Close the valve of the precursor bottle and allow the chamber to stand for 30 seconds (soaking time) before opening it to the pump again to ensure that the metal-containing precursor vapor reacts with the surface of the drug particles.

[0390] d. Thereafter, the ALD reactor is pumped for 9 seconds.

[0391] e. Repeat step ad above 20 times.

[0392] The chamber is then purged with a continuous stream of nitrogen to remove unreacted reagents and organic gases. Thereafter, steps (a) through (e) are essentially repeated, except that water is used as a second reagent to form a discrete alumina layer on the surface of the active ingredient particles. Subsequent purging pulses of nitrogen are then provided in a continuous stream to remove gaseous water and organic gases.

[0393] Repeat the above procedure nine times to form a total of ten initial aluminum oxide atomic layers.

[0394] Subsequently, in step (b) above, using diethylzinc (DEZ) and then water as precursors, three zinc oxide layers are applied by repeating the ALD step above to form a total of three zinc oxide layers. Then, an alumina layer is applied using the same precursor mentioned in section (b) above. This provides a total of four atomic layers of mixed oxide layers with an atomic ratio of 1:3 (Al₂O₃:ZnO).

[0395] This is repeated a total of 9 times to form a mixed oxide (1:3Al2O3:ZnO) layer with a total of 50 atomic layers (30 ZnO layers and 20 Al2O3 layers), including ten initial aluminum oxide atomic layers.

[0396] The powder was then removed from the ALD reactor and deagglomerated using an acoustic sieve (Tsutsui SW-20AT acoustic agitator sieve) with a sieve having a 20μm mesh size.

[0397] The resulting deagglomerated powder was re-sampled into the ALD reactor and coated with ten layers of pure alumina, followed by a layer of mixed oxide (1:3 Al₂O₃:ZnO) with a total of 50 atomic layers, following the same procedure. After the final deagglomeration step, the particles were coated with ten layers of pure alumina, followed by a layer of mixed oxide (1:3 Al₂O₃:ZnO), and finally post-treated with three layers of pure alumina with a total of 53 atomic layers. The deagglomeration and coating steps were repeated five times to produce a sample with a pure alumina underlayer, a finishing layer, and six discrete layers of mixed oxide coating (1:3 Al₂O₃:ZnO atomic ratio).

[0398] Therefore, the fourth batch contains 6 x 50 layers, which is 6 x (10 Al2O3 layers and 40 1:3 Al2O3:ZnO mixed oxide layers) and 3 Al2O3 (outer) layers.

[0399] Batch characterization

[0400] The particle size of a batch was determined by suspending the coated particles in a solution of 1% Span 85 (Sigma-Aldrich, Missouri, USA) in heptane (Merck, Germany). The particle size distribution, as measured by laser diffraction, is as follows:

[0401] Batch 3: %D(10): 3.7μm±9.9%; %D(50): 9.3μm±3.9%; %D(90): 21.3μm±5.9%;

[0402] Batch 4: Undetermined.

[0403] The drug loading of the coated batches was determined as follows: the sample was etched in an aqueous solution of 2M phosphoric acid (Merck, Germany) (PanReac, Spain) to dissolve the coating, and then diluted to 160 μg / mL. For batch 3, the sample diluent was a mixture of 10% mobile phase B (90% acetonitrile aqueous solution (Spectrascan, Scotland) + 0.1% trifluoroacetic acid (Sigma Aldrich, Germany)) 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 an HPLC system for quantification (Prominence-IHPLC-UV-DAD, Shimadzu, Japan) with a Kinetex F5 column. 150 × 4.6 mm, 2.6 μm particle size (Phenomenex, California, USA); Mobile phase A: 10% acetonitrile aqueous solution + 0.1% trifluoroacetic acid; Mobile phase B: 90% acetonitrile aqueous solution + 0.1% trifluoroacetic acid; Injection volume 10 μL, autosampler temperature 20 °C, oven temperature 25 °C; Gradient elution was used at a flow rate of 1.0 mL / min; UV absorbance was measured at 220 nm; Single-point calibration was used for determination; Liraglutide retention time was 6.8 min; Standard: 99.4% total liraglutide (as determined by assay), 99.1% purity (as determined by HPLC, Bachem AG, Switzerland).

[0404] The drug load of the third batch was 70.5±0.7%, and the drug load of the fourth batch was 46.8±0.2%.

[0405] To determine the integrity of the coating, two batches of samples were suspended in dimethyl sulfoxide (Rathburn, UK) at a concentration of 0.4 mg liraglutide per mL of solvent and rotated on a top-mounted stirrer for 3 hours. Intermittent samples were removed, centrifuged at 6000 rpm (EBA 20, Hettich, Germany) for 7 minutes, and the supernatant was diluted in mobile phase A (as above) before being injected into the HPLC system described above for quantification. After 3 hours, the results showed the release of 3.6% (batch 3) and 2.2% (batch 4) of liraglutide, indicating minimal defects in the coating, thus any burst release was expected to be low.

[0406] In vitro studies

[0407] 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 a blue-capped flask and incubated at 37 °C for up to 72 hours. Intermittent samples were taken and filtered through a 0.2 μm PES filter before analysis using HPLC with the same method described for determining drug loading. Results for batch 3 (circles) and batch 4 (squares) are shown in... Figure 5 middle.

[0408] Example 6

[0409] Formulation III of the invention

[0410] Suspensions of two coated liraglutide microparticles (batch 3 and batch 4, prepared according to the method described in Example 5 above) were prepared by adding an appropriate volume of veterinary hyonate (1 mM phosphate buffer containing 1 wt% hyaluronic acid solution, pH 7.4) to the test vials to achieve a liraglutide concentration corresponding to 10 mg / mL. The vials were then tapped at least 10 times to remove any material that might have precipitated at the bottom of the test vials. The mixture was then vortexed for approximately 60 seconds to ensure homogeneity of the suspension.

[0411] Example 7

[0412] In vivo study

[0413] On the day of application, it was administered by Ellegaard Göttingen Miniature Pig Breeding AG. MinipigsA / S (Denmark) provided nine male Göttingen SPF miniature pigs weighing approximately 10 kg. The animals were randomly divided into groups of three.

[0414] The suspension (prepared as described in Example 6 above) or a control liraglutide solution diluted in physiological saline (Novo Nordisk, Denmark) was administered subcutaneously as a single dose to three Göttingen miniature pigs in each dose group via a 23G needle into the left side of the neck of each pig. The miniature pigs receiving batch 3, batch 4, or control liraglutide solutions corresponded to groups 5, 6, and 7, respectively, as indicated in Table 3 below.

[0415] Table 3

[0416]

[0417] Blood samples for pharmacokinetic purposes were collected from all animals on Day 1 and in the following days. Blood samples were collected from all groups at the following time points: before treatment, and at 2, 4, 6, 12, 24, 48, 72, 96, and 144 hours after treatment. Additionally, blood was collected from animals in Groups 2 and 3 at 216, 360, 456, 552, and 648 hours after treatment (corresponding to Days 10, 16, 20, 24, and 28).

[0418] Approximately 3 mL of blood was drawn from the jugular vein / jugular trunk. The blood was collected into a vacuum blood collection tube containing K2EDTA as an anticoagulant. The vacuum blood collection tube was placed in ice water until centrifuged (10 minutes, 1270 G, +4°C). Each plasma sample was aliquoted into two aliquots of approximately 0.5 mL each and transferred within 90 minutes of collection to a cryovial provided by the sponsor and frozen at -70°C or on dry ice. The first set of samples was shipped on dry ice (approximately -70°C) with a thermometer to aid in analysis. The second set of samples was stored as a backup sample at -70°C or lower. The backup sample was shipped within one month of receiving the last set of original samples.

[0419] The plasma concentration of liraglutide was determined using LC-MS / MS. Study samples were prepared using a TECAN liquid handling robot by pipetting 35 μL of miniature pig plasma into 384-well plates and adding 35 μL of internal standard working solution. The 384-well plates were then shaken for 1 min. Protein precipitation solution was added, and the samples were shaken for 5 min and centrifuged. All samples were injected into a UPLC-MS / MS system. Separation was performed using a Waters ACQUITY Phenyl-Hexyl column (1.7 μm, 2.1 × 50 mm) at 60 °C, using 0.3% formic acid as mobile phase A (MP A) and acetonitrile as mobile phase B (MP B).

[0420] Non-compartmental analysis (NCA) was used to evaluate PK in Göttingen miniature pigs using Phoenix WinNonlin, version 8.3 (Certara, USA).

[0421] Nominal plasma sampling time points and nominal doses are used for non-compartmental PK analysis. Animal PK is calculated using the extravascular dosing option within WinNonlin. max Plasma concentrations below LLOQ that were previously observed are considered zero.

[0422] C was derived from observed plasma concentration data. max and T max AUC is assessed by integrating the plasma concentration versus time curve, using linear interpolation for increasing plasma levels and logarithmic interpolation (linear for the ascending segment and logarithmic for the descending segment) for decreasing plasma levels. For AUC... last Calculations were made up to the last point (AUC) showing the measurable plasma concentration. last The area of ​​).

[0423] result

[0424] Figure 6 The mean plasma concentrations of liraglutide over four weeks following a single subcutaneous administration of various formulations (n=3) are presented.

[0425] Plasma pharmacokinetic parameters are also presented as mean values ​​for each dose group (standard deviations are provided in parentheses) in Table 4 below, where C max and t max Corresponding to the definition as defined above, and:

[0426] ·'AUC 0-24h '-AUC from time 0 hours to time 24 hours'

[0427] ·'AUC last '-AUC from time 0 to the last detectable plasma concentration'

[0428] Table 4

[0429]

[0430] Compared to group 7, subcutaneous administration of the two coated liraglutide microparticle formulations (groups 5 and 6) showed an initial absorption period of liraglutide, followed by a slower and more prolonged absorption period spanning at least 28 days. For groups 5 and 6 (coated liraglutide microparticle formulations, dose 1.0 mg), the observed mean C0.0 of liraglutide was... max The results were significantly lower than those observed after subcutaneous administration of liraglutide solution (Group 7, dose 0.2 mg).

[0431] The plasma exposure to liraglutide in group 5 (batch 3) appeared to be higher than that in group 6 (batch 4), which may be due to the thicker coating formulation in group 6 compared to group 5.

[0432] In summary, groups 5 and 6 exhibited prolonged release curves, which is consistent with the higher C-values ​​observed after administration of liraglutide solution. max The subsequent rapid decline in plasma concentration was significantly different.

[0433] Example 8

[0434] Coated liraglutide microparticles

[0435] Liraglutide granules

[0436] Batches of spray-dried liraglutide particles with large sizes were prepared in a spray drying apparatus similar to that used in Example 1, using process parameters defined below. All other steps were performed according to the method described in Example 1.

[0437] For these formulations, liraglutide was dissolved in water for injection (WFI) to a concentration of 5% (w / w) during spray drying. 5g and 30g batches were produced in a small spray dryer, PROCEPT SD3, with equipment setup parameters as shown in Table 5. Process parameters Set values) and Table 6 ( Result parameter ranges during the process) Defined in [the document / reference].

[0438] Table 5

[0439]

[0440]

[0441] Table 6

[0442] Critical parameters Range Ambient temperature outside the chamber (°C) 60-70 Temperature inside the cyclone (°C) 30-40 Condenser temperature (°C) 0-4 Dosing speed (g / min) 4.0±1.0

[0443] Batch 5

[0444] For batch 5 (297_5_LIR), larger spray-dried liraglutide particles prepared as described above were used. The particle size of the larger spray-dried liraglutide particles as described above was determined as follows: %D(10): 3.1 μm ± 4.3%, %D(50): 10.9 μm ± 4.0%, %D(90): 30.5 μm ± 6.8%.

[0445] For batch 5, the large spray-dried liraglutide particles were first coated with ten layers of pure alumina using the following method:

[0446] a. Close the valve on the pipeline between the pump and the ALD reactor.

[0447] b. Then open the valve on the trimethylaluminum (TMA) precursor bottle for 1 second to allow the evaporated metal-containing precursor to fill the ALD reactor for 1 second.

[0448] c. Close the valve of the precursor bottle and allow the chamber to stand for 30 seconds (soaking time) before opening it to the pump again to ensure that the metal-containing precursor vapor reacts with the surface of the drug particles.

[0449] d. Thereafter, the ALD reactor is pumped for 9 seconds.

[0450] e. Repeat step ad above 20 times.

[0451] The chamber is then purged with a continuous stream of nitrogen to remove unreacted reagents and organic gases. Thereafter, steps (a) through (e) are essentially repeated, except that water is used as a second reagent to form a discrete alumina layer on the surface of the active ingredient particles. Subsequent purging pulses of nitrogen are then provided in a continuous stream to remove gaseous water and organic gases.

[0452] Repeat the above procedure nine times to form a total of ten initial aluminum oxide atomic layers.

[0453] Subsequently, in step (b) above, using diethylzinc (DEZ) and then water as a precursor, twenty zinc oxide layers are applied by repeating the ALD step above to form a total of twenty zinc oxide layers. This provides a multilayer structure of pure alumina and zinc oxide with a total of 30 atomic layers (Al2O3:ZnO), including ten initial alumina atomic layers.

[0454] The powder was then removed from the ALD reactor and deagglomerated using an acoustic sieve (Tsutsui SW-20AT acoustic agitator sieve) with a sieve opening size of 45 μm.

[0455] The resulting deagglomerated powder was re-loaded into the ALD reactor and coated with ten layers of pure alumina, followed by twenty layers of pure zinc oxide with a total of 30 atomic layers, following the same procedure. After the final deagglomeration step, the particles were coated with seven layers of pure alumina, followed by twenty layers of pure zinc oxide, and then post-treated with three layers of pure alumina to obtain a mixed oxide (Al₂O₃:ZnO:Al₂O₃) layer with a total of 30 atomic layers. The deagglomeration and coating steps were repeated four times to produce samples with five discrete layers of pure alumina and zinc oxide coating.

[0456] Therefore, the fifth batch contains 5 x 30 layers (10 Al2O3 layers and 20 ZnO layers) and 1 x 30 layers (7 Al2O3 layers, 20 ZnO layers and 3 Al2O3 layers).

[0457] Batch 6

[0458] For batch 6, spray-dried liraglutide particles as described in Example 1 were used.

[0459] For batch 6, the spray-dried liraglutide particles were first coated with ten layers of pure alumina using the following method:

[0460] a. Close the valve on the pipeline between the pump and the ALD reactor.

[0461] b. Then open the valve on the trimethylaluminum (TMA) precursor bottle for 1 second to allow the evaporated metal-containing precursor to fill the ALD reactor for 1 second.

[0462] c. Close the valve of the precursor bottle and allow the chamber to stand for 30 seconds (soaking time) before opening it to the pump again to ensure that the metal-containing precursor vapor reacts with the surface of the drug particles.

[0463] d. Thereafter, the ALD reactor is pumped for 9 seconds.

[0464] e. Repeat step ad above 20 times.

[0465] The chamber is then purged with a continuous stream of nitrogen to remove unreacted reagents and organic gases. Thereafter, steps (a) through (e) are essentially repeated, except that water is used as a second reagent to form a discrete alumina layer on the surface of the active ingredient particles. Subsequent purging pulses of nitrogen are then provided in a continuous stream to remove gaseous water and organic gases.

[0466] Repeat the above procedure nine times to form a total of ten initial aluminum oxide atomic layers.

[0467] Subsequently, in step (b) above, using diethylzinc (DEZ) and then water as precursors, three zinc oxide layers are applied by repeating the ALD step above to form a total of three zinc oxide layers. Then, an alumina layer is applied using the same precursor mentioned in section (b) above. This provides a total of four atomic layers of mixed oxide layers with an atomic ratio of 1:3 (Al₂O₃:ZnO).

[0468] This is repeated a total of 9 times to form a mixed oxide (1:3Al2O3:ZnO) layer with a total of 50 atomic layers (30 ZnO layers and 20 Al2O3 layers), including ten initial aluminum oxide atomic layers.

[0469] The powder was then removed from the ALD reactor and deagglomerated using an acoustic sieve (Tsutsui SW-20AT acoustic agitator sieve) with a sieve having a 20μm mesh size.

[0470] The resulting deagglomerated powder was re-sampled into the ALD reactor and coated with ten layers of pure alumina, followed by a layer of mixed oxide (1:3 Al₂O₃:ZnO) with a total of 50 atomic layers, following the same procedure. After the final deagglomeration step, the particles were coated with ten layers of pure alumina, followed by a layer of mixed oxide (1:3 Al₂O₃:ZnO) with a total of 50 atomic layers, following the same procedure. The deagglomeration and coating steps were repeated three times to produce a sample with a pure alumina underlayer and four discrete layers of mixed oxide coating (1:3 Al₂O₃:ZnO atomic ratio).

[0471] Therefore, the 6th batch contains 4 x 50 layers, which is 4 x (10 Al2O3 layers and 40 1:3 Al2O3:ZnO mixed oxide layers).

[0472] Batch 7

[0473] For batch 7, larger spray-dried liraglutide particles prepared as described above were used. The particle size of the larger spray-dried material was determined as follows: %D(10): 3.0 μm ± 1.2%, %D(50): 9.7 μm ± 2.4%, %D(90): 27.6 μm ± 6.1%.

[0474] For batch 7, the large spray-dried liraglutide particles were first coated with three layers of pure alumina using the following method:

[0475] a. Close the valve on the pipeline between the pump and the ALD reactor.

[0476] b. Then open the valve on the trimethylaluminum (TMA) precursor bottle for 1 second to allow the evaporated metal-containing precursor to fill the ALD reactor for 1 second.

[0477] c. Close the valve of the precursor bottle and allow the chamber to stand for 30 seconds (soaking time) before opening it to the pump again to ensure that the metal-containing precursor vapor reacts with the surface of the drug particles.

[0478] d. Thereafter, the ALD reactor is pumped for 9 seconds.

[0479] e. Repeat step ad above 20 times.

[0480] The chamber is then purged with a continuous stream of nitrogen to remove unreacted reagents and organic gases. Thereafter, steps (a) through (e) are essentially repeated, except that water is used as a second reagent to form a discrete alumina layer on the surface of the active ingredient particles. Subsequent purging pulses of nitrogen are then provided in a continuous stream to remove gaseous water and organic gases.

[0481] Repeat the above procedure twice to form a total of three initial aluminum oxide atomic layers.

[0482] Subsequently, in step (b) above, using diethylzinc (DEZ) and then water as precursors, three zinc oxide layers are applied by repeating the ALD step above to form a total of three zinc oxide layers. Then, an alumina layer is applied using the same precursor mentioned in section (b) above. This provides a total of four atomic layers of mixed oxide layers with an atomic ratio of 1:3 (Al₂O₃:ZnO).

[0483] This is repeated a total of 5 times to form a mixed oxide (1:3Al2O3:ZnO) layer with a total of 27 atomic layers (18 ZnO layers and 9 Al2O3 layers), including three initial aluminum oxide atomic layers.

[0484] The powder was then removed from the ALD reactor and deagglomerated using an acoustic sieve (Tsutsui SW-20AT acoustic agitator sieve) with a sieve opening size of 32 μm.

[0485] The resulting deagglomerated powder was re-sampled into the ALD reactor and coated with three layers of pure alumina, followed by a layer of mixed oxide (1:3 Al₂O₃:ZnO) with a total of 27 atomic layers, following the same procedure. After the final deagglomeration step, the particles were coated with three layers of pure alumina, followed by a layer of mixed oxide (1:3 Al₂O₃:ZnO) with a total of 27 atomic layers, following the same procedure. The deagglomeration and coating steps were repeated three times to produce a sample with a pure alumina underlayer and four discrete layers of mixed oxide coating (1:3 Al₂O₃:ZnO atomic ratio).

[0486] Therefore, the 7th batch contains 4 x 27 layers, which is 4 x (3 Al2O3 layers and 24 1:3 Al2O3:ZnO mixed oxide layers).

[0487] Batch 8

[0488] For batch 8, larger spray-dried liraglutide particles prepared as described above were used. The particle size of the larger spray-dried material was determined as follows: %D(10): 3.1 μm ± 4.3%, %D(50): 10.9 μm ± 4.0%, %D(90): 30.5 μm ± 6.8%.

[0489] For batch 8, the larger spray-dried liraglutide particles were first coated with three layers of pure alumina using the following method:

[0490] a. Close the valve on the pipeline between the pump and the ALD reactor.

[0491] b. Then open the valve on the trimethylaluminum (TMA) precursor bottle for 1 second to allow the evaporated metal-containing precursor to fill the ALD reactor for 1 second.

[0492] c. Close the valve of the precursor bottle and allow the chamber to stand for 30 seconds (soaking time) before opening it to the pump again to ensure that the metal-containing precursor vapor reacts with the surface of the drug particles.

[0493] d. Thereafter, the ALD reactor is pumped for 9 seconds.

[0494] e. Repeat step ad above 20 times.

[0495] The chamber is then purged with a continuous stream of nitrogen to remove unreacted reagents and organic gases. Thereafter, steps (a) through (e) are essentially repeated, except that water is used as a second reagent to form a discrete alumina layer on the surface of the active ingredient particles. Subsequent purging pulses of nitrogen are then provided in a continuous stream to remove gaseous water and organic gases.

[0496] Repeat the above procedure twice to form a total of three initial aluminum oxide atomic layers.

[0497] Subsequently, in step (b) above, using diethylzinc (DEZ) and then water as precursors, three zinc oxide layers are applied by repeating the ALD step above to form a total of three zinc oxide layers. Then, an alumina layer is applied using the same precursor mentioned in section (b) above. This provides a total of four atomic layers of mixed oxide layers with an atomic ratio of 1:3 (Al₂O₃:ZnO).

[0498] This is repeated a total of 5 times to form a mixed oxide (1:3Al2O3:ZnO) layer with a total of 27 atomic layers (18 ZnO layers and 9 Al2O3 layers), including three initial aluminum oxide atomic layers.

[0499] The powder was then removed from the ALD reactor and deagglomerated using an acoustic sieve (Tsutsui SW-20AT acoustic agitator sieve) with a sieve opening size of 45 μm.

[0500] The resulting deagglomerated powder was re-sampled into the ALD reactor and coated with three layers of pure alumina, followed by a layer of mixed oxide (1:3 Al₂O₃:ZnO) with a total of 27 atomic layers, following the same procedure. After the final deagglomeration step, the particles were coated with three layers of pure alumina, followed by a layer of mixed oxide (1:3 Al₂O₃:ZnO) with a total of 27 atomic layers, following the same procedure. The deagglomeration and coating steps were repeated three times to produce a sample with a pure alumina underlayer and four discrete layers of mixed oxide coating (1:3 Al₂O₃:ZnO atomic ratio).

[0501] Therefore, the 8th batch contains 4 x 27 layers, which is 4 x (3 Al2O3 layers and 24 1:3 Al2O3:ZnO mixed oxide layers).

[0502] Batch characterization

[0503] The particle size of a batch was determined by suspending the coated particles in a solution of 1% Span 85 (Sigma-Aldrich, Missouri, USA) in heptane (Merck, Germany). The particle size distribution, as measured by laser diffraction, is as follows:

[0504] Batch 5: %D(10): 7.2μm±3.9%; %D(50): 17.9μm±1.6%; %D(90): 36.2μm±2.1%;

[0505] Batch 6: Undetermined;

[0506] Batch 7: %D(10): 3.8μm ± 1.3%; %D(50): 11.2μm ± 1.9%; %D(90): 26.3μm ± 6.8%; and

[0507] Batch 8: %D(10): 4.5μm±5.3%; %D(50): 13.0μm±0.8%; %D(90): 30.0μm±0.5%.

[0508] The drug loadings of batches 5, 7, and 8 were determined as follows: Samples were etched in a solution of 88% water (PanReac, Spain) and 12% (v / v) phosphoric acid (Merck, Germany) to dissolve the coating, then diluted to 75 μg / mL with 75% water (Rathburn, UK) + 0.1% trifluoroacetic acid (Merck, Germany) and injected into an HPLC system (Prominence-IHPLC-UV-DAD, Shimadzu, Japan) with a Kinetex F5 column (150 x 4.6 mm, 150 × 4.6 mm, 2.6 μm particle size) for quantification; mobile phase A: comprising 0.1%... TFA was prepared as a 10% acetonitrile aqueous solution; mobile phase B consisted of 80% acetonitrile aqueous solution + 0.1% trifluoroacetic acid; injection volume was 2 μL; autosampler temperature was 20°C; oven temperature was 25°C; gradient elution was used at a flow rate of 1.0 mL / min; UV absorbance was measured at 220 nm; single-point calibration was used for determination; standard substance: 99.4% total liraglutide (as determined by assay), 99.1% purity (as determined by HPLC (Bachem AG, Switzerland)).

[0509] The drug loading of the 6th batch was determined as follows: the sample was etched to dissolve the coating in a solution of 43% acetonitrile (Spectrascan, Scotland), 45% water (PanReac, Spain), and 12% (v / v) phosphoric acid (Merck, Germany), then diluted to 160 μg / mL with 10% acetonitrile aqueous solution + 0.1% trifluoroacetic acid (Merck, Germany) (mobile phase A) and injected into a Kinetex C18 column for quantification. PS, 150×4.6mm, 2.6μm particle size (Phenomenex, California, USA) HPLC system (Prominence-IHPLC-UV-DAD, Shimadzu, Japan); mobile phase B: 80% acetonitrile aqueous solution + 0.1% trifluoroacetic acid; injection volume 10μL, autosampler temperature 15℃, oven temperature 25℃; gradient elution, flow rate 1.0mL / min; UV absorbance detection at 220nm; determination using single-point calibration; liraglutide retention time 6.8min; standard: 99.4% total liraglutide (as determined by assay), 99.1% purity (as determined by HPLC, Bachem AG, Switzerland).

[0510] Drug loading measured by HPLC-UV: 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).

[0511] To determine the integrity of the coating, the material was suspended in dimethyl sulfoxide (Rathburn, UK) at a concentration of 0.4 mg liraglutide per mL of solvent and rotated on a top-mounted stirrer for 3 hours. Intermittent samples were removed, centrifuged at 6000 rpm (EBA 20, Hettich, Germany) for 7 minutes, and the supernatant was diluted in mobile phase A (as above) before being injected into the HPLC system described above for quantification. After 3 hours, the results showed that batch 5 released 4.0% liraglutide, batch 6 released 4.6% liraglutide, batch 7 released 5.2% liraglutide, and batch 8 released 7.8% liraglutide, indicating minimal defects in the coating, thus any burst release was expected to be low.

[0512] In vitro studies

[0513] 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 a blue-capped flask and incubated at 37 °C for up to 72 hours. Intermittent samples were taken and filtered through a 0.2 μm PES filter before analysis using HPLC with the same method described for determining drug loading. Results for batch 5 (circular), batch 6 (square), batch 7 (diamond), and batch 8 (triangular) are shown in [Figure / Diagram / Illustration]. Figure 7 middle.

[0514] Example 9

[0515] Formulation IV of the invention

[0516] Suspensions of two types of coated liraglutide microparticles (batch 5, batch 6, batch 7, and batch 8, prepared according to the method described in Example 8 above) were prepared by adding an appropriate volume of veterinary hyonate (1 mM phosphate buffer containing 1 wt% hyaluronic acid solution, pH 7.4) to the test vials to achieve a liraglutide concentration corresponding to 10 mg / mL. The vials were then tapped at least 10 times to remove any material that might have precipitated at the bottom of the test vials. The mixture was then vortexed for approximately 60 seconds to ensure homogeneity of the suspension.

[0517] Example 10

[0518] In vivo study

[0519] According to Table 7 below, sixteen (16) male Sprague-Dawley rats were divided into four groups of four animals each. The animals were weighed and administered a single subcutaneous injection using a 23G needle. Blood samples were collected at the following time points after administration for plasma separation: 3 hours, 6 hours, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, 120 hours, 168 hours, 251 hours, 384 hours, 480 hours, 576 hours, and 672 hours. The administration site was visually inspected at blood sampling time points one hour and 24 hours after administration. The animals were euthanized after the final blood sample was collected.

[0520] Table 7

[0521]

[0522] a.Al+Zn refers to particles with a multilayer structure containing pure Al2O3 and ZnO.

[0523] b.1:3AZO refers to particles with mixed oxide layers (three ZnO layers and one Al2O3 layer).

[0524] For groups 1 through 3, blood samples (approximately 0.2 mL) were collected from the jugular vein into K2EDTA (dipotassium ethylenediaminetetraacetate) tubes at the following time points: 1 hour, 3 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 120 hours, 168 hours, 251 hours, 384 hours, 480 hours, 576 hours, and 672 hours after administration. For group 4, samples were collected at the following time points: 1 hour, 2 hours, 3 hours, 6 hours, 9 hours, 12 hours, 24 hours, and 48 hours after subcutaneous administration. The actual sampling time was recorded. Plasma was separated as soon as possible after blood sampling by centrifugation (1500 g, 4°C, for 10 minutes) and stored at -80°C until analysis.

[0525] After the study was completed, all plasma samples were deep-frozen on dry ice before being shipped for analysis. Animals were euthanized on the last day of the study.

[0526] Plasma concentrations of liraglutide were determined using LC-MS / MS. Study samples were prepared using an EVO-2 liquid handling robot (Teikken, Austria) by pipetting 35 μL of rat plasma into 96-well plates and adding 75 μL of internal standard working solution. The 96-well plates were then shaken for 15 minutes and centrifuged. All samples were then injected into a UPLC-MS / MS system (Waters Corporation, Massachusetts, USA). An ACQUITY UPLC Protein BEH C4 column was used. Separation was performed using a 1.7 μm, 2.1 mm x 50 mm (Waters Corporation, Massachusetts, USA) mobile phase A and acetonitrile as mobile phase B at 60 °C with a 0.3% formic acid aqueous solution.

[0527] Non-compartmental analysis (NCA) was performed using Phoenix WinNonlin version 8.3 software (Certara, USA). The pharmacokinetic (PK) analysis of liraglutide in plasma was evaluated. Cp was derived from the observed plasma concentration data. max and t max AUC is assessed by integrating the plasma concentration versus time curve, using linear interpolation for increasing plasma levels and logarithmic interpolation (linear for the ascending segment and logarithmic for the descending segment) for decreasing plasma levels. For AUC... ∞ Calculations were made up to the last point (AUC) showing the measurable plasma concentration. last The area of ​​the curve is calculated, and then the concentration in the last quantifiable sample and the first-order rate constant λ associated with the end of the curve are used. z Extrapolate to infinity. t 1 / 2,z From ln2 / λ z calculate.

[0528] result

[0529] Figure 8 and 9 It was shown that within one week after a single subcutaneous application of various formulations ( Figure 8 ) and within 42 days ( Figure 9 ) dose-normalized plasma concentration of liraglutide.

[0530] Plasma pharmacokinetic parameters are also presented in Table 8 below as means of the four groups (standard deviations are provided in parentheses), where the parameter definitions correspond to those above, and:

[0531] ·'Fr.Rel. 0-72h 'Is the percentage of the concentration released during the first 72 hours of the time curve, expressed as a percentage, up to infinity.

[0532] Table 8

[0533]

[0534]

[0535] from Figure 8 As can be seen, subcutaneous administration of the four formulations showed an initial absorption period of liraglutide, followed by a slower and longer absorption period spanning at least 7 days. Specifically, Figure 8 and 9 The formulation administered to group 11 showed higher release during the first 7 days, followed by lower release over 42 days, compared to groups 8 through 10 (which exhibited prolonged release exceeding 42 days). Group 8 (corresponding to batch 5, i.e., Al+Zn coated) showed lower C compared to group 11 (corresponding to batch 8, i.e., 1:3 AZO coated). max Both formulations are produced using 10.9 μm (% D50) liraglutide particles as raw materials.

Claims

1. A pharmaceutical formulation for treating metabolic disorders or symptoms, said pharmaceutical formulation comprising a plurality of particles suspended in a carrier system, said particles: (a) Having an average diameter based on weight, number, or volume between about 10 nm and about 700 μm; and (b) comprising a core containing at least one glucagon-like peptide-1 receptor agonist or a pharmaceutically acceptable salt thereof, the core being at least partially coated with an inorganic material comprising a mixture of the following: (i) Zinc oxide; and (ii) one or more other metal oxides and / or metal-like oxides. The atomic ratio ((i):(ii)) is at least about 1:10 and at most and includes about 10:

1.

2. The formulation according to claim 1, wherein the atomic ratio ((i):(ii)) is at least about 1:1 and at most and includes about 6:

1.

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

4. The formulation according to any one of the preceding claims, wherein the coated particles comprise more than one discrete layer surrounding the core.

5. The formulation according to claim 4, wherein at least one of the discrete layers is substantially composed of a single metal oxide and / or a metal-like oxide.

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

7. The formulation according to any one of claims 4 to 6, wherein the number of discrete layers is at most four single metal oxide and / or metal-like oxide layers.

8. The formulation according to any one of claims 4 to 7, wherein the single metal oxide and / or metal-like oxide is aluminum oxide.

9. The formulation according to any one of the preceding claims, wherein the core is substantially composed of at least one glucagon-like peptide-1 receptor agonist or a pharmaceutically acceptable salt thereof.

10. The formulation according to any one of the preceding claims, wherein the average diameter of the particles based on weight, quantity or volume is between about 1 μm and about 50 μm.

11. The formulation according to any one of the preceding claims, wherein more than one discrete oxide mixture layer is sequentially applied to the core.

12. The formulation according to claim 11, wherein between 3 and 10 of the discrete oxide mixture layers are applied.

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

14. The formulation according to any one of claims 4 to 13, wherein the maximum thickness of a single discrete oxide coating layer is about 1 percent of the average diameter of the core based on weight, quantity, or volume, including any other discrete layers previously applied to the core.

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

1.

16. The formulation according to any one of the preceding claims, wherein the one or more other metal oxides and / or metal-like oxides are selected from alumina and / or silicon dioxide.

17. The formulation according to any one of the preceding claims, wherein it is in the form of a sterile injectable dosage form.

18. The formulation according to claim 17, wherein it is in the form of a surgical application device capable of forming a reservoir formulation.

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

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

21. A method for preparing a formulation according to any one of the preceding claims, wherein the coated particles are prepared by applying a layer of mixed oxide coating material to the core and / or a previously coated core via atomic layer deposition.

22. The method according to claim 21, wherein: (i) Coating a solid core with a first discrete layer of a mixed oxide coating material; (ii) The coated core from step (i) is then subjected to a deagglomeration process step. (iii) Then the core from step (ii) is coated with a second discrete layer of mixed oxide coating material; (iv) Repeat steps (ii) and (iii) to obtain the desired number of discrete layers.

23. The method of claim 22, wherein the deagglomeration step occurring between coating applications comprises sieving.

24. The method of claim 23, wherein the screening comprises vibrating screening.

25. The method of claim 24, wherein the vibrating sieving includes controlling a vibrating probe coupled to the sieving.

26. The method of claim 23, wherein the sieving comprises acoustic sieving.

27. A method 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. An injectable dosage form comprising a formulation according to any one of claims 1 to 20, said formulation being contained in a reservoir connected to and / or associated with an injection device.

29. The dosage form according to claim 28 is a surgical application device for forming a reservoir preparation.

30. The dosage form according to claim 28 or claim 29, wherein it is an auto-injector.

31. The formulation according to any one of claims 1 to 20 or the dosage form according to any one of claims 28 to 30, for the treatment of metabolic disorders or symptoms.

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 preparation of a medicament for the treatment of metabolic disorders or symptoms.

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

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 comorbidities, or both.

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

36. The formulation, use, or method of use according to 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 daily or weekly injection of the at least one glucagon-like peptide-1 receptor agonist over the same time period.

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