New formulations containing biologically active drugs

Abstract

A pharmaceutical formulation useful for delivering high doses of a biologically active drug is provided, comprising a plurality of particles suspended in a carrier system, the particles (a) having an average diameter by weight, number, or volume that is from about 10 nm to about 700 μm, and (b) comprising a solid core comprising a biologically active drug at least partially coated with a coating of inorganic material, the formulation comprising a concentration of the biologically active drug of at least 10 mg / mL. Preferred biologically active drugs include immunoglobulins, monoclonal antibodies, antibody mimetics, cytokines and cytokine antagonists, and human peptide hormones.

Description

[Technical field]

[0001] The present invention relates to novel formulations for use in the field of drug delivery of biologically active drugs including, for example, immunoglobulins, antibodies, antibody mimetics, cytokines and cytokine antagonists, and human peptide hormones. [Background technology]

[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 common general knowledge.

[0003] In the field of drug delivery, biologically active drugs / active pharmaceutical ingredients (APIs), such as immunoglobulins, antibodies, and antibody analogs and mimetics, are increasingly proving useful for the treatment of a wide range of diseases and disorders.

[0004] Compared to other APIs, antibodies are less potent and may require injections daily, weekly, biweekly, or even longer intervals, typically at doses of 50-500 mg per injection.

[0005] One of the major problems with the use of biologically active drugs is the inability to form injectable formulations with high drug loadings in terms of the API, many of which form gels as concentrations increase.

[0006] For any sustained release composition, it is also very important that the release profile shows an initial minimal rapid release of active ingredient, i.e., high drug concentration in plasma immediately after administration. Such a "burst" release leads to undesirable high concentrations of active ingredient, which may be dangerous for drugs with narrow therapeutic windows. To ensure an optimal pharmacokinetic profile, it is also important to ensure that active ingredient is released at a desired and predictable rate in vivo after administration.

[0007] For example, in the case of injectable suspensions of biologically active pharmaceutical ingredients (APIs), such as protein formulations, including antibody formulations, due to the relatively low potency of antibodies and other proteins, it is often desirable to obtain high concentrations of the protein, and the associated need to provide high doses of the protein formulation.

[0008] High concentration protein formulations (HCPF) is the term used to refer to such formulations; although no clear definition of the concentration has been agreed upon, a typical range for monoclonal antibody (mAb) drugs is 50-150 mg / mL. HCPFs have undesirable properties that differ from those of lower protein concentration formulations, including a tendency for lower stability, a tendency for the compound to form gels, and an increased tendency for protein particulate formation.

[0009] Thus, there is a general need in the art for effective and / or improved drug transport and delivery systems for formulations containing high concentrations of biologically active drugs, such as antibody drugs.

[0010] Atomic layer deposition (ALD) is a technique used to deposit thin films on solid substrates, including a variety of materials, including organic, biological, polymeric, and especially inorganic materials such as metal oxides. It is a technique that enables atomic and near-atomic scale manufacturing (ACSM) of materials, structures, devices, and systems in versatile applications (see, for example, Zhang et al. Nanomanuf. Metrol. 2022, https: / / doi.org / 10.1007 / s41871-022-00136-8). Based on its self-limiting properties, ALD can achieve atomic-level thicknesses that are controlled only by adjusting the number of growth cycles. Moreover, multiple layers can be deposited, and the properties of each layer can be customized at the atomic level.

[0011] Because of its atomic level control, ALD is used, for example, as a key technique in the production of next generation semiconductors, or for the atomic level synthesis of advanced catalysts, as well as for the precise fabrication of nanostructures, nanoclusters, and single atoms (see, e.g., Zhang et al., supra).

[0012] This technique is usually carried out at low pressure and high temperature. Film coatings are produced by alternately exposing a solid substrate in an ALD reactor chamber to reactants vaporized in the gas phase. The substrate can be a silicon wafer, a granular material, or small particles (e.g., microparticles or nanoparticles).

[0013] The coated substrate is protected from chemical reactions (decomposition) and physical changes by the solid coating. ALD can potentially also be used to control the release rate of substrate materials in a solvent, which could potentially be used to formulate active pharmaceutical ingredients.

[0014] In ALD, a first precursor, which may be metal-containing, is delivered to the ALD reactor chamber (in a so-called "precursor pulse") and forms a monolayer of atoms or molecules adsorbed on the surface of the substrate. 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 and forms a monolayer, for example of a metal oxide, on the substrate surface. A subsequent purge pulse is followed by a further pulse of the first precursor, thus initiating a new cycle of the same events (a so-called "ALD cycle").

[0015] The thickness of the film coating is controlled, among other things, by the number of ALD cycles performed.

[0016] In a typical ALD process, only monolayers of atoms or molecules are produced during a single cycle, so that no discernible physical interfaces form between these monolayers, resulting in essentially continuous bands across the surface of the substrate.

[0017] International Patent Application WO2014 / 187995 describes a process in which several ALD cycles are performed, after which the resulting coated substrate is periodically removed from the reactor and a redispersion / agitation step is performed to present new surfaces available for precursor adsorption.

[0018] The agitation step is performed to solve a problem observed mainly with nano- and microparticles, namely, particle agglomeration during the ALD coating process, which results in the formation of "pinholes" due to the contact points between such particles. The redispersion / agitation step is performed by placing the coated substrate in water and sonicating, which results in deagglomeration and the destruction of the contact points between the individual particles of the coated active material.

[0019] The particles were then reloaded into the reactor and the steps of ALD coating the powder and deagglomerating the powder were repeated three times (for a total of four successive cycles). This process has been shown to enable the formation of coated particles that are, to a large extent, pinhole-free (see also Hellrup et al, Int. J. Pharm., 529, 116 (2017)).

[0020] In the particular case of injectable suspensions, it is also important to ensure that the suspended particle size is controlled so that they can be injected through a needle: if large aggregated particles are present, not only will they block the needle through which the suspension is injected, but they will not form a stable suspension in the injection fluid (i.e., they will instead tend to sink to the bottom of the injection fluid).

[0021] We provide novel injectable biologically active drug formulations in which ALD is used to coat drug microparticles with an inorganic coating layer in which the coated particles are suspended in a vehicle, the drug being one or more biologically active drugs. These formulations generate favorable pharmacokinetic profiles by releasing the active ingredient over an extended period of time, providing therapeutically effective levels of drug in the systemic circulation without a significant initial burst effect.

[0022] Also provided are pharmaceutical formulations obtained by the methods disclosed herein that allow for high concentrations of an API, such as HCPF, while reducing the negative attributes associated with formulations with high concentrations of the API. Summary of the Invention

[0023] According to a first aspect of the present invention, there is provided a pharmaceutical formulation useful for delivering high doses of a biologically active drug (API) comprising a plurality of particles suspended in a carrier system, said particles comprising: (a) have an average diameter based on weight, number, or volume that is from about 10 nm to about 700 μm; (b) comprises a solid core containing a biologically active drug at least partially coated with a coating of inorganic material. This formulation contains a concentration of the API of at least about 10 mg / mL and is hereinafter referred to as the "formulation of the present invention." [Brief description of the drawings]

[0024] [Figure 1] FIG. 1 shows the absorbance measured at 450 nm in a UV-VIS plate reader after ELISA on coated and uncoated spray-dried microparticles containing monoclonal antibody ATH3G10. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] It will be well understood by those skilled in the art that the term "solid" includes any form of matter that retains its shape and density when unconfined and / or whose molecules are generally as tightly packed together as the repulsive forces between them allow. The solid core has at least a solid outer surface onto which a layer of coating material can be deposited. The interior of the solid core may also be solid or alternatively hollow. For example, if the particles are spray dried before being placed in a reaction vessel, they may be hollow due to the spray drying technique.

[0026] The formulation of the present invention is preferably a pharmaceutical formulation, in which case the formulation may contain a pharmacologically effective amount of a biologically active drug. Furthermore, the solid core preferably contains the biologically active drug.

[0027] In this regard, the solid core may consist essentially of or comprise a biologically active drug or agent (which drug or "agent" may hereinafter be interchangeably referred to as "active pharmaceutical ingredient (API)" and / or "active ingredient"). Biologically active drugs also include biopharmaceuticals and / or biologics. Biologically active drugs may also comprise a mixture of different APIs, either as different API particles or as particles containing multiple APIs. It is contemplated that the biologically active drug according to the present invention is selected from the group comprising immunoglobulins, antibodies, monoclonal antibodies, and antibody mimetics.

[0028] The drug loading contained in the formulations of the present invention can be determined as dry drug loading, which refers to the drug loading as a weight percent of the particles contained in the formulation, and suspended drug loading, which refers to the solids content as a weight percent of the formulations of the present invention in the form of a suspension.

[0029] The formulations of the present invention comprise a high dry drug loading, such as, for example, about 20% by weight or more, such as, for example, about 30% by weight or more, such as, for example, about 40% by weight or more, such as, for example, about 50% by weight or more, such as, for example, about 60% by weight or more, such as, for example, about 70% by weight or more, such as, for example, about 80% by weight or more, such as, for example, about 90% by weight or more, such as, for example, about 95% by weight or more, such as at least about 99% by weight.

[0030] The formulation of the present invention comprises a suspension having a solid content of about 20% or more, for example, about 30% or more, for example, about 40% or more, for example, about 50% or more, for example, about 60% or more, for example, about 70% or more, for example, about 80% or more.The solid content then includes the API as well as the coating used on the particles of the formulation.Why is the total drug load in the suspension affected by the dry drug load of the particles?Therefore, the total drug load may be lower than the solid content of the suspension.

[0031] As described in detail below, the formulations for the present invention may be in the form of a liquid, sol, paste, or gel that can be administered via a surgical administration device to form a depot formulation. It is contemplated that increasing the solids content of the formulations of the present invention will increase the likelihood that the formulation will be in the form of an injectable paste.

[0032] A formulation of the invention may comprise a concentration of API of at least about 10 mg / mL, such as at least about 25 mg / mL, such as at least about 50 mg / mL, such as at least about 100 mg / mL, such as at least about 150 mg / mL, such as at least about 200 mg / mL, at least about 250 mg / mL, such as at least about 300 mg / mL, such as at least about 350 mg / mL, such as at least about 400 mg / mL, such as at least about 450 mg / mL, such as at least about 500 mg / mL.

[0033] Desirable APIs and formulations of the present invention may include immunoglobulins such as those present with ATC (anatomical therapeutic chemistry) codes within J06BA and J06BB, such as IgG for use in replacement and / or replacement therapy in immunodeficiency disorders and immune regulation. Such replacement or replacement therapy is used in the treatment of various neurological disorders, Guillain-Barre syndrome, Kawasaki disease, and in patients suffering from cancer, including lymphomas such as B-cell non-Hodgkin's lymphoma, Hodgkin's lymphoma, leukemias such as chronic lymphocytic leukemia (CLL), and / or patients treated for cancer by undergoing chemotherapy treatments that reduce the number of B cells or destroy B cells. Examples of such treatments include, for example, treatment with rituximab.

[0034] Immune globulin replacement / replacement therapy is also used in immune thrombocytopenia and autoimmune hemolytic anemia, especially in patients who have not responded positively to other treatments.

[0035] Immune globulin replacement / replacement therapy is also used in patients with primary immunodeficiencies such as common variable immunodeficiency (CVID) and X-linked agammaglobulinemia.

[0036] Non-limiting examples of immunoglobulins that may be used in accordance with the present invention include immunoglobulin, normal human, for extravascular administration (J06BA01), immunoglobulin, normal human, for intravascular administration (J06BA02), anti-D(rh) immunoglobulin (J06BB01), tetanus immune globulin (J06BB02), varicella / zoster immune globulin (J06BB03), hepatitis B immune globulin (J06BB04), rabies immune globulin (J06BB05), rubella immune globulin (J06BB06), vaccinia immune globulin (J06BB07), staphylococcus immune globulin (J06BB08), cytomegalovirus immune globulin (J06BB09), and / or erythropoietin immune globulin (J06BB10). (J06BB09), diphtheria immune globulin (J06BB10), hepatitis A immune globulin (J06BB11), encephalitis, tick-borne immune globulin (J06BB12), pertussis immune globulin (J06BB13), measles immune globulin (J06BB14), parotitis immune globulin (J06BB15), palivizumab (J06BB16), motavizumab (J06BB17), raxibacumab (J06BB18), bezlotoxumab (J06BB21), obiltoxaximab (J06BB22), anthrax immune globulin (J06BB19), combinations (J06BB30), or mixtures thereof.

[0037] The desired API and formulation of the present invention may comprise a monoclonal antibody. Non-limiting examples of (monoclonal) antibodies that may be used according to the present invention include edrecolomab (L01XC01), rituximab (L01XC02), trastuzumab (L01XC03), gemtuzumab ozogamicin (L01XC05), cetuximab (L01XC06), bevacizumab (L01XC07), panitumumab (L01XC08), catumaxomab (L01XC09), ofatumumab (L01XC10), ipilimumab (L01XC11), brentuximab vedotin (L01XC12), pertuzumab (L01XC13), trastuzumab Emtansine (L01XC14), obinutuzumab (L01XC15), dinutuximab beta (L01XC16), nivolumab (L01XC17), pembrolizumab (L01XC18), blinatumomab (L01XC19), ramucirumab (L01XC21), necitumumab (L01XC22), elotuzumab (L01XC23), daratumumab (L01XC24), mogamulizumab (L01XC25), inotuzumab Ozogamicin (L01XC26), Olaratumab (L01XC27), Durvalumab (L01XC28), Bermekimab (L01XC29), Avelumab (L01XC31), Atezolizumab (L01XC32), Cemiplimab (L01XC33), Moxetumomab passudotox (L01XC34), Tafasitamab (L01XC35), Enfortumab vedotin (L01XC36), Polatuzumab vedotin (L01XC37), Isatuximab (L01XC38), Belantamab mafodotin (L01XC39), Dostarlimab (L01XC40), Trastuzumab deruxtecan (L01XC41), phosphorylcholine monoclonal antibodies, bispecific T cell engagers (BiTEs) such as blinatumomab, solitomab, AMG330, MT112, MT111, BAY2010112, MEDI-565, or any mixture thereof.

[0038] The desired API and formulation of the present invention may include antibody mimetics. Non-limiting examples of antibody mimetics that may be used according to the present invention include affibody molecules (such as ABY-025), affilins (such as SPVF2801), affimers, affitins, alphabodies (such as CMPX-1023), anticalins, avimers, designed ankyrin repeat proteins (DARPins such as MP0112), finomers, Kunitz domain peptides (such as Ecallantide (Kalbitor)), adnectins and monobodies (such as Pegdinetanib (Angiocept)), nanoCLAMPs, single domain antibodies such as camelid antibodies, and VNARs derived from cartilaginous fishes (IgNARs). NAR fragments, bivalent single domain antibodies (such as caplacizumab (Cablivi)), as well as armadillo repeat proteins (here designed armadillo repeat proteins), peptide aptamers, and knottins, or any mixtures thereof.

[0039] The desired API and formulation of the present invention may include human peptide hormones. Non-limiting examples of human peptide hormones that may be used in accordance with the present invention include amylin, anti-Mullerian hormone, adiponectin, adrenocorticotropic hormone, angiotensinogen, angiotensin, antidiuretic hormone, atrial natriuretic peptide, brain natriuretic peptide, calcitonin, cholecystokinin, corticotropin releasing hormone, cortistatin, enkephalin, endothelin, erythropoietin, follicle stimulating hormone, galanin, gastric inhibitory polypeptide, gastrin, ghrelin, glucagon, glucagon-like peptide-1, gonadotropin releasing hormone, growth hormone releasing hormone, hepcidin, human chorionic gonadotropin, human placental lactogen, growth hormone, inhibin, insulin, insulin-like growth factor, leptin, lipotropin, luteinizing hormone, melanocyte stimulating hormone, motilin, orexin, osteocalcin, oxytocin, pancreatic po peptide, parathyroid hormone, pituitary adenylate cyclase-activating peptide, prolactin, prolactin-releasing hormone, relaxin, renin, secretin, somatostatin, growth hormone-inhibiting hormone, growth hormone-releasing inhibitory hormone, somatotropin-releasing inhibitory factor, somatotropin-releasing inhibitory hormone, thrombopoietin, thyrotropin, thyrotropin-releasing hormone, vasoactive intestinal peptide, guanylin / uro These include guanylin, tetracosactide, mecaselmin, somapasitan, pegvisomant, vasopressin, desmopressin, terlipressin, lypressin, ornipressin, argipressin, demoxitocin, carbetocin, gonadorelin, nafarelin, histrelin, octreotide, lanreotide, vapreotide, pasireotide, ganirelix, cetrorelix, elagolix, relugolix, teriparatide, and elcatonin.

[0040] Desirable APIs and formulations of the present invention may include cytokines and analogs, including recombinant cytokines. Non-limiting examples of cytokines and analogs include IL-1 receptor antagonists, anakinra, IL-2, IL-7, IL-15, IL-21, TNF-alpha, interferon-gamma, IFN-alpha, pifonakin, movenakin, adargilkin alfa, aldesleukin, sermoleukin, denilkin diftitox, pegaldesleukin, teceleukin, tucotuzumab sermoleukin, daniprestim, muprestim, binetrakin, atexakin alfa, emoctakine, ilodecakin, oprelvekin, edekin alfa, syntredekin budox, ivoctadekin, cytokines developed into protein therapeutics (e.g., bone morphogenetic proteins (BMPs), erythropoietin (EPO), granulocyte colony stimulating factor (G-CSF), interferon alpha, interferon beta, IL-11, interferon gamma, and the like.

[0041] The solid core of the formulations of the present invention comprises a biologically active drug, and in this regard may consist essentially of one or more biologically active drugs and / or may comprise one or more biologically active drugs together with other excipients and / or other active ingredients.

[0042] "Consisting essentially of" a biologically active drug includes that the solid core contains essentially only the biologically active drug, i.e., is free of non-biologically active substances such as excipients, carriers (discussed below), and other active substances. This means that the core may contain less than about 5%, such as less than about 3%, such as less than about 2%, such as less than about 1% of such other excipients and / or active substances.

[0043] Alternatively, the core containing a biologically active drug may contain such an active ingredient mixed with one or more pharmaceutical ingredients, which may include pharma- ceutically acceptable excipients such as adjuvants, diluents, or carriers, and / or may include other biologically active ingredients.

[0044] Non-biologically active adjuvants, diluents and carriers which may be used in the cores coated according to the invention may include pharma- ceutically acceptable substances which are soluble in water, such as carbohydrates, e.g., sugars such as lactose and / or trehalose, and sugar alcohols such as mannitol, sorbitol and xylitol, or pharma-ceutically acceptable inorganic salts such as sodium chloride. Preferred carrier / excipient materials include sugars and sugar alcohols.

[0045] The biologically active drug may be presented in a crystalline, partially crystalline, and / or amorphous state. Regardless of the physical form, the biologically active drug may be in or converted to a solid state at about room temperature (e.g., about 18° C.) and about atmospheric pressure. The active agent (and optionally other pharmaceutical ingredients described herein) should also remain in solid form while being coated, for example, in the ALD reactor, and should not physically or chemically degrade to any significant extent (i.e., about 10% w / w or less) while being coated or after being covered by the coating material.

[0046] The formulations of the present invention contain a pharmacologically effective amount of one or more biologically active drugs. Preferably, the solid core of the formulations of the present invention contains said pharmacologically effective amount of said one or more biologically active drugs.

[0047] The term "pharmacologically effective amount" refers to an amount of one or more biologically active agents capable of producing a desired physiological change (such as a therapeutic effect) in a treated patient, whether administered alone or in combination with another active ingredient. Such biological or medical response in a patient, or such effect, may be subjective (i.e., the subject gives an indication of or feels an effect), may be objective (i.e., measurable by some test or marker), including at least partial alleviation of the symptoms of the disease or disorder being treated, or a cure or prevention of said disease or disorder.

[0048] Therefore, the dose of one or more biologically active drugs that can be administered to a patient must be sufficient to affect a therapeutic response over a reasonable and / or relevant time frame.Those skilled in the art recognize that the selection of the exact dose and composition and the most suitable delivery regimen is influenced not only by the nature of the biologically active drug, but also by, among others, the pharmacological properties of the formulation, the route of administration, the nature and severity of the condition being treated, the physical condition and mental acuity of the recipient, and the age, condition, weight, sex, and response of the patient being treated, the stage / severity of the disease, and the genetic differences between patients.

[0049] The dosage of the one or more biologically active drugs may also be determined by the timing and frequency of administration. In any event, a physician or other skilled in the art will be able to routinely determine the actual dosage of the one or more biologically active drugs that is most suitable for an individual patient.

[0050] Upon injection, the formulations of the present invention provide a depot formulation in which one or more biologically active drugs are released over an extended period of time, which may be at least about 3 days, e.g., about 5 days, or about 7 days, and up to about a year, e.g., about 3 weeks (e.g., about 2 weeks or about 4 weeks), or about 12 weeks (e.g., about 10 weeks or about 14 weeks).

[0051] Thus, a suitable dose of one or more biologically active drugs in a formulation of the invention will be an exposure (e.g., AUC ) that provides at least the same therapeutic effect as that obtained for current, commercially available subcutaneous and / or intravenous injections of one or more biologically active drugs used in clinical practice. last (area under the plasma concentration versus time curve to the last detectable concentration over an extended period of time), more preferably AUC ∞ The present invention may provide plasma concentration-time (AUC) values ​​that provide a linear correlation between the plasma concentration and time (AUC) of the analyte concentration in the blood and the plasma concentration versus time curve (AUC) up to time infinity.

[0052] The formulations of the present invention are designed to provide a therapeutically effective dose of one or more biologically active drugs administered by injection or infusion for 7 consecutive days, which is comparable to the total exposure (AUC ∞) that is less than or equal to 100% of the AUC of one or more biologically active drugs in plasma over any one of the above periods. ∞ It may be possible to provide exposure to.

[0053] The solid biologically active drug-containing core of the formulation of the present invention is provided in the form of nanoparticles, or more preferably microparticles, preferably having an average diameter by weight, number, or volume of about 50 nm (e.g., about 100 nm, e.g., about 250 nm) to about 30 μm, e.g., about 500 nm to about 100 μm, more specifically, about 1 μm to about 50 μm (e.g., about 25 μm, e.g., about 20 μm).

[0054] As used herein, the term "weight-based average diameter" is understood by those skilled in the art to include the average particle size being characterized and defined from a particle size distribution by weight, i.e., a distribution in which the existing fraction (relative amount) in each size class is defined as a weight fraction obtained, for example, by sieving (e.g., wet sieving). As used herein, the term "number-based average diameter" is understood by those skilled in the art to include the average particle size being characterized and defined from a particle size distribution by number, i.e., a distribution in which the existing fraction (relative amount) in each size class is defined as a number fraction measured, for example, by microscopy. As used herein, the term "volume-based average diameter" is understood by those skilled in the art to include the average particle size being characterized and defined from a particle size distribution by volume, i.e., a distribution in which the existing fraction (relative amount) in each size class is defined as a volume fraction measured, for example, by laser diffraction. Those skilled in the art will also understand that there are other suitable ways of expressing average diameter, such as area-based average diameter, and these other expressions of average diameter are interchangeable with those used herein. Other instruments well known in the art may be used to measure particle size, for example those sold by Malvern Instruments, Ltd (Worcestershire, UK) and Shimadzu (Kyoto, Japan).

[0055] The particles may be spherical, i.e., they have an aspect ratio of less than about 20, more preferably less than about 10, such as less than about 4, especially less than about 2, and / or may have a variation in radius (measured from the center of gravity to the particle surface) of at least about 90% of the particles of not more than about 50% of the average value, such as not more than about 30% of that value, for example not more than about 20% of that value.

[0056] Nevertheless, the present invention also allows for coating of particles of any shape. For example, irregularly shaped (e.g., "raisin" shaped), needle-shaped, flaky or rectangular particles can be coated. For non-spherical particles, the size can be indicated as the corresponding spherical particle size of, for example, the same weight, volume or surface area. Hollow particles, as well as particles with pores, gaps, etc., such as fibrous or "entangled" particles, can also be coated according to the present invention.

[0057] The particles may be obtained in a form suitable for them to be coated, or in that form, for example, by a particle size reduction process (e.g., crushing, shearing, milling, or grinding to a particular weight-based average diameter (as described above), for example, by utilizing wet grinding, dry grinding, air jet milling (including cryogenic micronization), ball milling such as planetary ball milling, as well as end runner mills, roller mills, vibratory mills, hammer mills, roller mills, fluid energy mills, pin mills, and the like. Alternatively, the particles may be directly prepared to a suitable size and shape, for example, by spray drying, freeze drying, spray freeze drying, vacuum drying, spray drying including the use of supercritical fluids, precipitation, or other top-down methods (i.e., reducing large particle sizes, for example, by grinding), or bottom-up methods (i.e., increasing small particle sizes, for example, by sol-gel techniques). Alternatively, nanoparticles may be made by well-known techniques such as gas condensation, attrition, chemical precipitation, ion implantation, pyrolysis, hydrothermal synthesis, and the like.

[0058] The particles may need to be washed and / or cleaned to remove impurities that may result from their production, and then dried (depending on the method by which the particles containing the cores are initially provided). Drying may be carried out by many techniques known to those skilled in the art, including evaporation, spray drying, vacuum drying, freeze drying, fluidized bed drying, microwave drying, IR radiation, drum drying, etc. Once dried, the cores may then be deagglomerated by grinding, screening, milling, and / or dry sonication. Alternatively, the cores may be treated to remove any volatile materials that may be absorbed on their surface, for example by exposing the particles to vacuum and / or elevated temperatures.

[0059] The surface of the core may be chemically activated prior to application of the first layer of coating material, for example by treatment with hydrogen peroxide, ozone, a free radical-containing reactant, or by applying a plasma treatment to create free oxygen radicals on the surface of the core, which may create favorable adsorption / nucleation sites on the core for the ALD precursors.

[0060] Preferred methods for applying coatings to cores containing biologically active agents include gas phase techniques such as ALD or related techniques such as atomic layer epitaxy (ALE), molecular layer deposition (MLD, a technique similar to ALD except that molecules (usually organic molecules) are deposited with each pulse instead of atoms), molecular layer epitaxy (MLE), chemical vapor deposition (CVD), atomic layer CVD, molecular layer CVD, physical vapor deposition (PVD), sputtering PVD, reactive sputtering PVD, evaporation PVD and binary reaction sequence chemistry. ALD is the preferred coating method according to the present invention.

[0061] When ALD is used, the coating material may be prepared by feeding a precursor into the ALD reactor chamber (in a so-called "precursor pulse") to form a monolayer of atoms or molecules adsorbed on the surface of the particle. A second precursor is then pulsed into the reactor and reacts with the first precursor to form a monolayer of the compound on the substrate surface. A subsequent purge pulse is followed by a further pulse of the first precursor, thus initiating a new cycle of the same events (a so-called "ALD cycle").

[0062] In most cases, the first of the successive reactions involves some functional group or free electron pair or radical on the surface to be coated, such as a hydroxy group (-OH) or a primary or secondary amino group (-NH2 or -NHR, where R is an aliphatic group, e.g., an alkyl group). Each reaction is advantageously carried out separately under conditions such that essentially all excess reagents and reaction products are removed before carrying out the next reaction.

[0063] In ALD, the layer of coating material may be applied at a process temperature of about 20° C. to about 100° C., for example, about 40° C. to about 100° C. The optimal process temperature depends on the reactivity of the precursor. It is preferred to use lower temperatures, such as about 30° C. to about 100° C. In particular, temperatures of about 20° C. to about 80° C., for example, about 30° C. to about 70° C., for example, about 40° C. to about 60° C., for example, about 50° C., are employed.

[0064] The core may be coated with one or more separate individual layers of an inorganic coating as defined herein. Preferably, two or more separate layers, coatings or shells (these terms are used interchangeably herein) are applied (i.e., "separately applied") to the solid core containing the biologically active agent.

[0065] Such "separate application" of a "separate layer, coating, or shell" means that a solid core is coated with a first layer of coating material, which layer is formed by two or more (e.g., a plurality or set of) cycles as described herein, each cycle producing a monolayer of coating material, and then the resulting coated core is subjected to some form of sieving step, such as a vibratory sieving technique, step, or process, as described herein.

[0066] In other words, a "vapor deposition (e.g., ALD) cycle" can be repeated several times to provide a "vapor deposition (e.g., ALD) set" of cycles, which can consist of, for example, 10, 25, or 100 cycles. However, after this series of cycles, the coated cores are subjected to some form of deagglomeration process, which is followed by a further series of cycles.

[0067] This process can be repeated as many times as necessary, so that the number of separate layers of coating material produced by multiple successive cycles in the final coating corresponds to the number of these intermittent deagglomeration steps, with a final mechanical deagglomeration occurring prior to application of the final layer (series of cycles) of coating material.

[0068] The terms "deagglomeration" and "disaggregation" are used interchangeably when referring to coated particles, and deagglomeration of coated particle agglomerates is preferably accomplished by mechanical sieving techniques.

[0069] The coated cores may be subjected to the aforementioned deagglomeration process internally without being removed from the apparatus in a continuous process. Such a process may include forcing the solid product mass formed by coating the cores through a sieve disposed within the reactor, configured to deagglomerate particle agglomerates upon forcing the coated cores by a forcing means applied within the reactor before being subjected to a second and / or further coating. This process may be continued as many times as necessary and / or appropriate before applying a final coating, as described herein.

[0070] Placing the sieve within the reaction vessel means that the coating can be applied by a continuous process without the need to remove the particles from the reactor. There is therefore no need for manual handling of the particles and no external machinery is required to deagglomerate agglomerated particles. This not only significantly reduces the time over which the coating process is carried out, but also makes it more convenient and reduces the risk of harmful (e.g. toxic) materials being handled by personnel. It also makes the process more repeatable by limiting manual handling and reduces the risk of contamination.

[0071] Alternatively and / or preferably, the coated cores may be removed from the coating apparatus, such as an ALD reactor, and then subjected to an external deagglomeration step, for example as described in International Patent Application WO2014 / 187995. Such an external deagglomeration step may include agitation, such as wet or dry sonication, or, preferably, sieving the resulting solid product mass discharged from the reactor, for example by passing through a sieve or mesh, to deagglomerate the particles, for example as described below, before returning the particles to the coating apparatus for the next coating step. Again, this process may be continued as many times as necessary and / or appropriate before applying the final coating.

[0072] In the external deagglomeration process, deagglomeration can alternatively be performed 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 for the cores) prior to the next coating step.

[0073] However, in such external processes, the deagglomeration step may include one or more sieving steps, which may include jet sieving, hand sieving, vibratory sieve shaking, horizontal sieve shaking, tap sieving, or (preferably) sonic sieving as described below, or a similar process involving any combination of these sieving steps. Manufacturers of suitable sonic sifters include Advantech Manufacturing, Endecott, and Tsutsui.

[0074] Vibratory screening techniques may involve vibratory passing of the solid product mass formed by coating the cores through a screen located internal or (preferably) external (i.e., outside) of the reactor, configured to deagglomerate any particle agglomerates during the vibratory passing of the coated cores prior to subjecting them to a second and / or further layer(s) of coating material, the process being repeated as many times as necessary and / or appropriate prior to applying a final layer of coating material.

[0075] The vibrationally passing means may include a vibration motor coupled to the sieve. The vibration motor is configured to vibrate and / or rotate when powered. For example, the vibration motor may be a piezoelectric vibration motor including a piezoelectric material that changes shape when an electric field is applied as a result of the inverse piezoelectric effect. The change in shape of the piezoelectric material induces acoustic or ultrasonic vibration of the piezoelectric vibration motor.

[0076] Alternatively, the vibration motor may be an eccentric rotating mass (ERM) vibration motor that includes a mass that is rotated when power is provided to the motor. The mass is eccentric from the axis of rotation, and rotation of the mass causes the motor to become unbalanced and vibrate and / or rotate. Additionally, an ERM vibration motor may include multiple masses located at different positions relative to the motor. For example, an ERM vibration motor may include an upper mass and a lower mass, each located at opposite ends of the motor. By varying each mass and its angle relative to the other mass, the vibration and / or rotation of the ERM vibration motor can be varied.

[0077] The vibration motor is coupled to the sieve in a manner such that when power is supplied, vibration and / or rotation of the motor is transmitted to the sieve.

[0078] The sieve and vibrating motor may be suspended from a mount (e.g., a frame that can be placed on the floor, etc.) via a suspension means such that the sieve and motor are free to vibrate relative to the mount without vibrations being substantially transmitted to or damped by the mount. This allows the vibrating motor and sieve to vibrate and / or rotate without obstruction and also reduces noise generated during the vibratory sieving process. The suspension means may include one or more springs or bellows (i.e., air cushions or equivalent cushioning means) that couple the sieve and / or motor to the mount. Manufacturers of vibrating sieves or sifters suitable for carrying out such processes include, for example, Russell Finex, SWECO, Filtra Vibracion, VibraScreener, Gough Engineering, and Farley Greene.

[0079] Preferably, the vibrating sieving technique further comprises controlling a vibrating probe coupled to the sieve. The vibrating probe may be controlled to vibrate the sieve at a frequency other than the frequency of the vibrations caused by the vibration motor. Preferably, the vibrating probe vibrates the sieve at a higher frequency than the vibrations caused by the vibration motor, more preferably the frequency is in the ultrasonic range.

[0080] Providing additional vibration to the sieve by the vibrating probe reduces the occurrence of sieve clogging, reduces the likelihood of the sieve being overloaded, and reduces the time required to clean the sieve mesh.

[0081] Preferably, the vibratory sieving technique involves sieving the coated particles at a throughput of at least 1 g / min. More preferably, the vibratory sieving technique involves sieving the coated particles at a throughput of 4 g / min or more.

[0082] The throughput depends on the sieve mesh area, sieve mesh size, particle size, particle cohesion, and particle static properties. Combining several of these features allows for much higher throughput. Thus, the vibratory sieving technique may more preferably include sieving coated particles at throughputs of up to 1 kg / min or more.

[0083] Any one of the above throughputs represents a significant improvement over the use of known mechanical sieving or screening techniques. For example, we have found that sonic sieving involves 15 minutes of sieving, with a 15 minute cooling time required to store the equipment. To sieve 20 g of coated particles, 9 sets of 15 minutes of active sieving time were required, i.e., a total time (including cooling) of 255 minutes. By comparison, by using the aforementioned vibration sieving technique, 20 g of coated particles can be continuously sieved in a maximum of 20 minutes, or more preferably in only 5 minutes or less.

[0084] The mesh size of the sieve may be determined such that the ratio of the size of the sieved or sonic sieved particles to the mesh size of the sieve is about 1:>1, preferably about 1:2, and optionally about 1:4. The mesh size of the size may range from about 20 μm to about 100 μm, preferably about 20 μm to about 60 μm.

[0085] Suitable sieve meshes may include perforated plates, microplates, grids, diamonds, threads, polymers or wires (woven wire sieves), but are preferably formed from a metal such as stainless steel.

[0086] Surprisingly, the use of stainless steel mesh within the vibratory sieving technique is gentle on particle coatings, as is the use of softer polymeric sieving mesh as part of a mechanical sieving technique such as sonic sieving.

[0087] Also, a known problem with sifting powders is the generation of potentially dangerous static electricity. Steel mesh has the advantage of removing static electricity from the powder, whereas polymer mesh does not. Polymer mesh must be used with a sonic sifter.

[0088] Furthermore, the mesh size of known sonic sifters is limited to about 100 μm because the sound waves move through the mesh rather than vibrating it. That limitation does not exist for the vibratory sifting technique, since it does not rely on sound waves to create vibrations in the sieve. Thus, the vibratory sifting technique described herein allows for sifting of larger particles than would be possible if alternative mechanical sifting techniques were used.

[0089] When the sieve is located external to the reactor (i.e., outside the reactor), the process of making the coated cores of the formulation of the present invention includes discharging the coated particles from the vapor deposition reactor before subjecting the coated particles to agitation, and reintroducing the deagglomerated coated particles into the vapor deposition reactor before applying at least one additional layer of coating material to the reintroduced particles.

[0090] The inventors have found that applying separate layers of coating material after external deagglomeration results in visible and discernible interfaces that can be observed by analyzing the coated particles according to the invention, e.g., by TEM, as regions of high electron transparency. In this regard, the thickness of the layer between the interfaces directly corresponds to the number of cycles performed in each series within the ALD reactor and between the individual external agitation steps.

[0091] In ALD coating processes, such distinct physical interfaces are typically more difficult to observe because the coating occurs at the atomic level.

[0092] Without being limited by theory, it is believed that removing the coated particles from the vacuum conditions of the ALD reactor and exposing the newly coated surface to air leads to structural reorganization due to relaxation and restructuring of the outermost atomic layers. Such a process is believed to involve a reorganization of the atoms at (and near) the surface, driven by a thermodynamic tendency to reduce the free energy of the surface.

[0093] Furthermore, surface adsorption of species (e.g., hydrocarbons, which are always present in air) may contribute to this phenomenon, as well as reactions of coatings formed with hydrocarbons, as well as surface modification by atmospheric oxygen, etc. Thus, chemical analysis of such interfaces may contain traces of contaminants not originating from the coating process, such as ALD, or core materials, such as APIs, that form part of the core.

[0094] Whether performed inside or outside the reactor, the particle agglomerates are preferably broken down by forcing them through a sieve, thus separating the agglomerates into individual particles or agglomerates of a desired and predetermined size (thereby achieving deagglomeration). With regard to the latter, in some cases, the individual primary particle sizes are so small (i.e., <1 μm) that it is not possible to achieve "complete" deagglomeration (i.e., the agglomerates are broken down into individual particles). Instead, deagglomeration is achieved by breaking down the larger agglomerates into smaller agglomerates of secondary particles of the desired size, as determined by the size of the sieve mesh. The smaller agglomerates are then coated by gas-phase techniques to form fully coated "particles" in the form of small agglomerate particles. Thus, the term "particles", when referring to deagglomerated and coated particles in the context of the present invention, refers to both individual (primary) particles and agglomerated (secondary) particles of the desired size.

[0095] In either case, the desired particle size (whether it be individual particles or agglomerates of the desired size) is maintained, and furthermore, continued application of the gas phase coating mechanism to the particles after such deagglomeration by sieving will form a complete coating on the particles, thus meaning that fully coated particles (individual or agglomerates of the desired size) are formed.

[0096] The above repeated coating and deagglomeration process, whether performed inside or outside the reactor, may be performed at least once, preferably twice, more preferably three times, such as four, five times, more particularly six times, such as seven times, and up to about 100 times, such as up to about 50 times, such as up to about 40 times, up to about 30 times, for example, 2 to 20 times, such as 3 to 15 times, for example, 10 times, such as 9 or 8 times, more preferably 6 or 7 times, especially 4 or 5 times.

[0097] Whether performed inside or outside the reactor, it is preferred that at least one screening step is performed, which further preferably comprises a vibratory screening step as described above. It is further preferred that the final screening step comprises a vibratory screening step performed prior to application of the final layer (series of cycles) of coating material. However, it is further preferred that two or more (including each) of the screening steps comprise the vibratory screening techniques, steps or processes described herein.

[0098] The preferred repetition of these steps further benefits the improved throughput of any vibratory screening technique.

[0099] The total thickness of the coating (meaning all separate layers / coatings / shells) is in the range of about 0.5 nm to about 2 μm on average.

[0100] The minimum thickness of each individual layer / coating / shell is in the range of about 0.1 nm on average (eg, about 0.5 nm, such as about 0.75 nm, eg 1 nm).

[0101] The maximum thickness of each individual layer / coating / shell will depend on the size of the core (initially) and then the size of the core with any previously applied coatings, and can be, on average, about 1 / 100 of the average diameter (i.e., average diameter based on weight, number, or volume) of the core, or of the cores to which the previously applied coatings have been applied.

[0102] Preferably, for particles having an average diameter of about 100 nm to about 1 μm, the total coating thickness should be, on average, about 1 nm to about 5 nm, for particles having an average diameter of about 1 μm to about 20 μm, the coating thickness should be, on average, about 1 nm to about 10 nm, and for particles having an average diameter of about 20 μm to about 700 μm, the coating thickness should be, on average, about 1 nm to about 100 nm.

[0103] It has been found that applying a coating / shell followed by one or more deagglomeration steps, such as sonication, can result in wear, pinholes, breaks, gaps, cracks, and / or voids (hereinafter "cracks") in the layer / coating because the coated particles are essentially "bonded" or "glued" more tightly upon application of a thicker coating, which can expose the core containing the biologically active component to elements as deagglomeration occurs.

[0104] For example, if it is intended to provide the sample in a suspension prior to administration to a patient, it is necessary to provide deaggregated primary particles without pinholes or cracks in the coating, which would result in an undesirable initial peak (burst) in the plasma concentration of the active ingredient immediately after administration.

[0105] We have discovered that performing one or more of the deagglomeration steps described herein results in particles with significantly fewer pinholes, gaps, or cracks in the final layer of coating material, resulting in particles that are not only completely covered with that layer / coating, but are also coated in a manner that allows the particles to be readily deagglomerated (e.g., using non-invasive techniques such as vortexing) in a manner that does not disrupt the layer of coating material formed prior to and / or during pharmaceutical formulation.

[0106] In this regard, the (e.g., inorganic) material coating typically completely surrounds, encloses, and / or encapsulates the solid core containing the biologically active drug, thus minimizing the risk of an initial drug concentration burst resulting from direct drug contact with a solvent in which the associated active ingredient is soluble, which may include not only bodily fluids but also any medium in which such coated particles may be suspended prior to injection.

[0107] Thus, in a further embodiment of the present invention there is provided a particle as disclosed herein, wherein the coating surrounding, surrounding and / or encapsulating the core covers at least about 50%, such as at least about 65%, for example at least about 75%, such as at least about 80%, more particularly at least about 90%, such as at least about 91%, for example at least about 92%, such as at least about 93%, for example at least about 94%, such as at least about 95%, for example at least about 96%, such as at least about 97%, for example at least about 98%, such as at least about 99%, for example approximately or about 100% of the surface of the solid core, wherein the coating essentially completely surrounds, surrounds and / or encapsulates the core.

[0108] As used herein, the term "essentially completely coating, completely surrounding, enclosing, and / or encapsulating the core" refers to coverage of at least about 98%, or at least about 99%, of the surface of the solid core.

[0109] Alternatively, the processes described herein may result in deagglomerated coated particles that are essentially free of such cracks through which the active ingredient may be released in an uncontrolled manner.

[0110] Although some minor cracks may appear in the coating without affecting its essential function in terms of controlling release, in further embodiments, particles as disclosed herein are provided in which at least about 90% of the particles do not exhibit cracks in the coating surrounding, surrounding and / or encapsulating the core. In one embodiment, 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 approximately 100% of the particles do not exhibit such cracks.

[0111] Alternatively, a coating being "essentially free of cracks" means that less than about 1% of the surface of the coated particle contains wear, pinholes, breaks, gaps, cracks and / or voids through which the active ingredient may potentially be exposed (e.g., to the elements).

[0112] The layers of coating material can be collectively essentially uniform in thickness across the surface area of ​​the particle. By "essentially uniform" thickness is meant that the degree of variation in coating thickness of at least about 10%, e.g., about 25%, e.g., about 50% of the coated particles present in the compositions of the invention is no more than about ±20% (including no more than ±50%) of the average thickness as measured by TEM.

[0113] Inorganic coating materials may include one or more metals or metalloids, or may include one or more metal-containing or metalloid-containing compounds, such as metals or metalloids, oxides, nitrides, sulfides, selenides, carbonates, and / or other ternary compounds, etc. Metals and metalloids, hydroxides, especially oxides, especially metal oxides are preferred.

[0114] Metals that may be mentioned include alkali metals, alkaline earth metals, noble metals, transition metals, post-transition metals, lanthanides, etc. Metals and metalloids that may be mentioned include aluminum, titanium, magnesium, iron, gallium, zinc, zirconium, niobium, hafnium, tantalum, lanthanum and / or silicon, more preferably aluminum, titanium, magnesium, iron, gallium, zinc, zirconium and / or silicon, in particular aluminum, silicon, titanium and / or zinc.

[0115] As noted above, because the compositions made by the processes of the present invention include two or more separate layers of inorganic coating material, the nature and chemical composition of those layers can vary from layer to layer.

[0116] Individual layers may also include mixtures of two or more inorganic materials, such as metal oxides or metalloid oxides, and / or may include multiple layers or composites of different inorganic or organic materials to modify the properties of the layer.

[0117] Coating materials that may be mentioned include aluminum oxide (Al2O3), titanium dioxide (TiO2), iron oxide (Fe x O y , for example FeO and / or Fe2O3 and / or Fe3O4), gallium oxide (Ga2O3), magnesium oxide (MgO), zinc oxide (ZnO), niobium oxide (Nb2O5), hafnium oxide (HfO2), tantalum oxide (Ta2O5), lanthanum oxide (La2O3), zirconium dioxide (ZrO2), and / or silicon dioxide (SiO2). Preferred coating materials include aluminum oxide, titanium dioxide, iron oxide, gallium oxide, magnesium oxide, zinc oxide, zirconium dioxide, and silicon dioxide. More preferred coating materials include iron oxide, titanium dioxide, zinc sulfide, more preferably zinc oxide, silicon dioxide, and / or aluminum oxide.

[0118] The layer of coating material (individually or collectively) may consist essentially of (e.g., greater than about 80%, e.g., greater than about 90%, e.g., about 95%, e.g., about 98%) iron oxide, titanium dioxide, or more preferably zinc oxide, silicon oxide, and / or aluminum oxide.

[0119] Thus, there is further provided a method of preparing a plurality of coated particles according to the invention, the coated particles being made by applying, by vapor deposition techniques, precursors of at least two metal and / or metalloid oxides forming a mixed oxide onto a solid core and / or a previously coated solid core. Precursors for forming metal or metalloid oxides often include water, oxygen, ozone, and / or oxygen precursors such as hydrogen peroxide, and metal and / or metalloid compounds, typically organometallic or organometalloid compounds.

[0120] Non-limiting examples of precursors are as follows: Precursors of zinc oxide can be water and di-C1-C5 alkyl zinc such as diethyl zinc. Precursors of aluminum oxide can be water and tri-C1-C5 alkyl aluminum such as trimethyl aluminum. Precursors of silicon oxide (silica) can be water as oxygen precursor, and silane, alkyl silane, amino silane, and orthosilicate tetraethyl ester. Precursors of iron oxide include oxygen, ozone and water as oxygen precursor, and di-C1-C5 alkyl iron, dicyclopropyl iron, and FeCl3. It will be understood that a person skilled in the art will recognize which precursors are suitable for the purposes disclosed herein.

[0121] It is further preferred that the inorganic coating material comprises a mixture of: (i) zinc oxide (ZnO); (ii) one or more other metal and / or metalloid oxides. The atomic ratio ((i):(ii)) is between at least about 1:6 and at most about 6:1.

[0122] Preferably, the atomic ratio ((i):(ii)) is between at least about 1:1 and at most about 6:1.

[0123] Coatings comprising a mixture of zinc oxide and one or more other metal and / or metalloid oxides are hereinafter referred to as "mixed oxide" coatings or coating materials.

[0124] Thus, the biologically active drug-containing core may be coated with a coating material comprising a mixture of zinc oxide and one or more other metal and / or metalloid oxides in an atomic ratio of zinc oxide to other oxides of at least about 1:6 (e.g., at least about 1:4, e.g., at least 1:2), preferably at least about 1:1 (e.g., at least about 1.5:1, e.g., at least about 2:1) (including at least about 2.25:1, e.g., at least about 2.5:1 (e.g., at least about 3.25:1 or at least about 2.75:1 (including 3:1)), and up to (i.e., up to) about 6:1 (including up to about 5.5:1), or up to about 5:1, e.g., up to 4.5:1 (including up to about 4:1) (e.g., up to about 3.75:1).

[0125] To produce a mixed oxide coating in which the atomic ratio of zinc oxide to one or more other metal and / or metalloid oxides is between (for example) about 1:1 and up to about 6:1, one skilled in the art will understand that for every one ALD cycle (i.e., monolayer) of the other oxide, about one to about six ALD cycles of zinc oxide must also be deposited. For example, if a mixed oxide coating with a 3:1 atomic ratio (zinc:other oxide) is to be formed, three zinc-containing precursor pulses are followed by a second precursor pulse each to form three monolayers of zinc oxide, followed by one pulse of the other metal and / or metalloid-containing precursor, followed by a second precursor pulse to form one monolayer of the other metal and / or metalloid oxide. Alternatively, six monolayers of zinc oxide may be followed by two monolayers of the other oxide, or any other combination, to provide an overall atomic ratio of about 3:1. In this regard, the order of pulses to produce the relevant oxides is not important, so long as the resulting atomic ratios are ultimately within the relevant range.

[0126] We have found that when coatings including zinc oxide are applied using ALD at low temperatures, such as from about 50° C. to about 100° C., the coating material is largely crystalline in nature (unlike other coating materials such as aluminum oxide and titanium oxide, which form amorphous layers).

[0127] Without being limited by theory, it is understood that because zinc oxide is crystalline, when zinc oxide alone is employed as a coating material, interfaces may form between adjacent crystals of zinc oxide deposited by ALD, which may allow penetration of the carrier system, medium, or solvent in which the zinc oxide is partially soluble (e.g., an aqueous solvent system) after suspension therein. It is believed that this may cause dissolution that is too rapid for the depot-forming composition that is intended to be made.

[0128] We have found that these problems can be alleviated by making mixed oxide coatings as described herein. In particular, we have found that these problems can be alleviated by making a mixture of two or more metal and / or metalloid oxide (mixed oxide) coatings as described herein. In particular, by forming a mixed oxide coating as described herein, which may be composed primarily, but not entirely, of zinc oxide, it was possible to coat the active ingredient with a coating that appears to be essentially amorphous, or a composite between crystalline and amorphous materials, and / or that may reduce the ingress of injection vehicles such as water. In this regard, it appears that the presence of the aforementioned perceived interfaces can be reduced or avoided entirely by employing the mixed oxide aspects of the invention in either a heterogeneous manner (where other oxides "fill" the gaps formed by the interfaces) or a homogeneous manner (where a true composite of mixed oxide materials is formed during deposition in a manner in which interfaces are potentially avoided in the first place).

[0129] In addition to the inorganic coatings employed in the formulations of the present invention, other coating materials, which may be pharma- ceutically acceptable and essentially non-toxic coating materials, may also be additionally applied between the separate inorganic coatings described herein (e.g., during a separate deagglomeration step) and / or while the inorganic coatings herein are being applied. Such materials may include multiple layers or composites of the coating materials described herein and one or more different inorganic or organic materials to modify the properties of the layers.

[0130] The additional coating material may include organic or polymeric materials such as polyamides, polyimides, polyureas, polyurethanes, polythioureas, polyesters, or polyimines. The additional coating material may also include hybrid materials (such as between organic and inorganic materials), including materials that are combinations between a metal or another element and an alcohol, a carboxylic acid, an amine, or a nitrile. However, it is preferred that the coating material comprises an inorganic material.

[0131] The vapor deposition reactor chamber used may optionally and / or preferably be a stationary vapor deposition reactor chamber. The term "stationary" in the context of a vapor deposition reactor chamber will be understood to mean that the reactor chamber remains stationary during use to carry out a vapor deposition technique, excluding negligible movement and / or vibration, such as caused by associated machinery, for example.

[0132] Furthermore, a so-called "stop-flow" process can be employed. Using the stop-flow process, the first precursor is fed into the reactor chamber and allowed to contact the core in the reactor chamber for a predetermined period of time (which can be considered as a soak time) before the first precursor is purged from the reactor chamber. During the predetermined period of time, there is preferably a substantial absence of a pump that can cause gas flow and / or a substantial absence of mechanical agitation of the core.

[0133] The employment of a stop-flow process can enhance the uniformity of the coating by allowing each gas to conform and diffuse into high aspect ratio substrates such as powders. The advantage can be even more pronounced when using slower reacting precursors, since the precursors are given more time to react on the surface. This can be particularly evident when depositing mixed oxide coatings according to the invention. For example, when depositing mixed zinc oxide / aluminum oxide coatings as described below, we have found that zinc-containing precursors, such as diethylzinc (DEZ), have a lower reaction probability towards the surface of the substrate than aluminum-containing precursors, such as trimethylaluminum (TMA).

[0134] In addition to producing coatings with good shell integrity and more controlled release profiles, the employment of such a stopped-flow process may improve the ability to achieve specific coating compositions.

[0135] For example, as noted above, when attempting to employ gas phase techniques to produce a coating in which the atomic ratio between zinc and aluminum in the resulting shell is 3:1, it was found that a ratio much closer to 3:1 could be achieved using a stopped-flow process than when depositing the material using a continuous flow of precursor.

[0136] Preferably and / or optionally, a "multi-pulse" technique may be employed to deliver the first precursor, the second precursor, or both precursors to the reactor chamber.

[0137] Using such multi-pulse techniques, each precursor may be delivered to the reactor chamber as multiple "sub-pulses," each lasting a short period of time, such as, for example, 1 second to about 1 minute (depending on the size and nature of the vapor deposition reactor), rather than as one continuous pulse. The precursor may be contacted with the core in the reactor chamber for a predetermined period of time, such as, for example, about 1-500 seconds, about 2-250 seconds, about 3-100 seconds, about 4-50 seconds, or about 5-10 seconds, such as 9 seconds, after each sub-pulse. Again, depending on the size and nature of the vapor deposition reactor, this time may extend up to several minutes (e.g., up to about 30 minutes). The introduction of the sub-pulses, followed by a period of soak time, may be repeated a predetermined number of times, such as, for example, about 5-1000 times, about 10-250 times, or about 20-50 times, in a single step.

[0138] Although the coated particles according to the present invention are essentially free of the aforementioned cracks in the applied coating through which the active ingredient would potentially be exposed (e.g., to the elements), two further optional steps may be applied to the coated particles before subjecting them to further processing of the pharmaceutical formulation.

[0139] The first optional step, after the final deagglomeration step described above, may involve the application of a final overcoating layer, where the thickness of that outer "overcoating" layer / coating, or "sealing shell" (these terms are used interchangeably herein) should be thinner than any previously applied separate layer / coating / shell (or "subshell").

[0140] Thus, the thickness can be about 0.7 times (e.g., about 0.6 times) or less on average the thickness of the widest previously applied subshell. Alternatively, the thickness can be about 0.7 times (e.g., about 0.6 times) or less on average the thickness of the last subshell applied and / or about 0.7 times (e.g., about 0.6 times) or less on average the average thickness of all previously applied subshells. The thickness can range from about 0.3 nm to about 10 nm on average for particles up to about 20 μm. For larger particles, the thickness can be about 1 / 1000 or less on average the average diameter based on weight, number, or volume of the coated particles.

[0141] The role of the sealing shell is to provide a "sealing" overcoating layer to the particles, covering their cracks, resulting in particles that are not only completely covered with their sealing shell, but also covered in a manner that allows the particles to be easily deagglomerated (e.g., using non-invasive techniques such as vortexing) in a manner that does not disrupt the subshell formed underneath before and / or during pharmaceutical formulation.

[0142] For reasons described herein, the sealing shell preferably does not include zinc oxide, although it may include silicon dioxide or, more preferably, aluminum oxide.

[0143] A second optional step may include subjecting the few remaining particles having broken and / or cracked shells / coatings to a process in which all particles are suspended in a solvent (wherein the biologically active drug is soluble, e.g., with a solubility of at least about 0.1 mg / mL, while the least soluble material of the inorganic coating is insoluble, e.g., with a solubility of about 0.1 μg / mL or less), followed by separation of the solid particles from the solvent, e.g., by centrifugation, sedimentation, flocculation, and / or filtration, thereby ensuring that primarily intact particles remain.

[0144] The above optional step, as previously discussed, provides a means of further potentially reducing the possibility of a (possibly) undesirable initial peak (burst) in plasma concentration of the active ingredient.

[0145] At the end of the process, the coated particles can be dried using one or more of the techniques previously described for drying the cores. Drying can occur in the absence or presence of one or more pharma- ceutically acceptable excipients (e.g., sugars or sugar alcohols).

[0146] Alternatively, at the end of the process, the separated particles may be resuspended in a solvent (e.g., water, with or without the presence of one or more pharma- ceutically acceptable excipients as defined herein) for subsequent storage and / or administration to a patient.

[0147] Prior to applying the first layer of coating material or between successive coatings, the cores and / or partially coated particles may be subjected to one or more alternative and / or preliminary surface treatments. In this regard, one or more intermediate layers comprising a different material (i.e., other than an inorganic material) may be applied to the relevant surface, for example, to protect the cores or partially coated particles from undesired reactions with precursors during the coating step / deposition process, to enhance coating efficiency, or to reduce agglomeration.

[0148] The intermediate layer may contain one or more surfactants, for example, to reduce the agglomeration of the particles to be coated and provide a hydrophilic surface suitable for subsequent coating. In this regard, suitable surfactants include well-known nonionic, anionic, cationic, or zwitterionic surfactants, such as the Tween series (e.g., Tween 80). Alternatively, the core may be subjected to a preliminary surface treatment if the active ingredient used as part of (or as) the core is susceptible to react with one or more precursor compounds that may be present in the gas phase during the coating (e.g., ALD) process.

[0149] Alternatively, application of an "intermediate" layer / surface treatment of this nature may alternatively be accomplished by liquid phase non-coating techniques followed by freeze drying, spray drying, or other drying methods to provide the particles with a surface layer onto which a coating material may then be applied.

[0150] The outer surface of the particles of the formulations of the invention may also be derivatized or functionalized with chemical compounds or moieties that enhance, for example, targeted delivery of the particles within a patient to which the nanoparticles are administered, for example, by attaching one or more chemical compounds or moieties to the outer surface of the final layer of coating material. Such compounds may be organic molecules (e.g., PEG) polymers, antibodies or antibody fragments, or receptor binding proteins or peptides, etc.

[0151] Alternatively, the moiety can be an anchoring group, such as a moiety containing silane functionality (see, e.g., Herrera et al, J. Mater. Chem., 18, 3650 (2008) and US 8,097,742). Another compound (e.g., a desired targeting compound) can be attached to such an anchoring group by covalent or non-covalent bonds (including hydrogen bonds or van der Waals bonds), or a combination thereof.

[0152] The presence of such an anchoring group may provide a versatile tool for targeted delivery to specific sites in the body. Alternatively, compounds such as PEG may be used to allow the particles to circulate longer in the bloodstream and avoid accumulation in the liver or spleen (the body's natural mechanisms for eliminating particles, potentially preventing delivery to diseased tissues).

[0153] The core coated with an inorganic coating, whether in the form of a separate, distinct layer, coating, or shell, as defined herein, is hereinafter referred to as the "coated particle of the formulation of the present invention."

[0154] Pharmaceutical (or veterinary) formulations comprising the compositions of the invention may contain different types of particles, for example particles with different functionalization (as described above), particles of different sizes and / or layers of inorganic coating material 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 may be controlled (e.g., varied or extended) over a particular period of time.

[0155] The formulations of the invention may be administered systemically, for example, by injection or infusion, intravenously or intra-arterially (including by intravascular or other perivascular devices / formulations (e.g., stents)), intramuscularly, intraosseously, intracerebrally, intraventricularly, intrasynovially, intrasternally, intrathecally, intralesional, intracranial, intratumoral, cutaneously, intradermally, subcutaneously, transdermally, in a pharmaceutical (or veterinarily) acceptable dosage form.

[0156] Preparation of the formulations of the present invention involves incorporating the coated particles described herein into a suitable pharma- ceutically acceptable aqueous carrier system, which may be accomplished taking into account the intended route of administration and standard pharmaceutical practice. Suitable excipients must therefore be chemically inert to the active agent employed and have no adverse side effects or toxicity under the conditions of use. Such pharma-ceutically acceptable carriers may also provide immediate or modified release of the biologically active agent from the particles of the formulations of the present invention.

[0157] For parenteral administration, such as subcutaneous and / or intramuscular injection, the compositions made by the process of the present invention may be in sterile injectable and / or infusible forms, for example, in the form of a sterile oily or preferably aqueous suspension of the compositions made by the process of the present invention.

[0158] Sterile aqueous suspensions of particles of the formulation of the invention can be formulated according to techniques known in the art. The aqueous medium should contain at least about 50% water, but may include other aqueous excipients such as Ringer's solution, and may also include polar co-solvents (e.g., ethanol, glycerol, propylene glycol, 1,3-butanediol, polyethylene glycols of various molecular weights, and tetraglycols), viscosity increasing or thickening agents (e.g., carboxymethylcellulose, microcrystalline cellulose, hydroxypropyl methylcellulose, hydroxyethyl cellulose, ethylhydroxyethyl cellulose, sodium starch glycolate, poloxamers such as poloxamer 407, polyvinylpyrrolidone, cyclodextrins such as hydroxypropyl-β-cyclodextrin, polyvinylpyrrolidone, and polyethylene glycols of various molecular weights), surfactants / wetting agents to achieve homogenous suspension (e.g., sorbitan esters, sodium lauryl sulfate, monoglycerides, polyoxyethylene esters, polyoxyethylene alkyl ethers, polyoxylglycerides, and preferably Tweens (Polysorbates), e.g., Tween 80 and Tween 20). Preferred ingredients include isotonicity modifiers (e.g., sodium lactate, dextrose, especially sodium chloride), pH adjusters and / or buffers (e.g., citric acid, sodium citrate, especially phosphate buffers such as disodium hydrogen phosphate hydrate, sodium phosphate, sodium dihydrogen phosphate monohydrate, and combinations thereof, which may be employed in combination with standard inorganic acids and bases such as hydrochloric acid and sodium hydroxide), and other ingredients such as mannitol, croscarmellose sodium, and hyaluronic acid.

[0159] The oily or oil-based carrier system may include one or more pharma- ceutically or veterinarily acceptable liquid lipids, which may include fixed oils such as mono-, di-, or triglycerides, including Miglyol (e.g., 812N), propylene glycol dicaprylocaprate (Miglyol 840, C8 / C10 ester), tricaprylin (Miglyol Oil), Gelcire 43 / 01, Corisolv GTA, Labrafil. Carrier systems may also include 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 pharma- ceutically acceptable oils, such as olive oil, peanut oil, soybean oil, corn oil, cottonseed oil, sesame oil, castor oil, oleic acid, and polyoxyethylated versions thereof (e.g., sorbitan trioleate, lauroglycol 90, capryol PGMC, PEG-60 hydrogenated castor oil, polyoxyl 35 castor oil).More preferred carrier systems include mono-, di-, and / or triglycerides, including alkyl chain triglycerides (e.g., C6-C 12 Medium chain triglycerides, such as alkyl chain triglycerides, are most preferred.

[0160] Such injection suspensions may be formulated according to techniques well known to those skilled in the art by employing suitable dispersing or wetting agents (eg, Tween, such as Tween 80) and suspending agents.

[0161] The formulations of the present invention may further be formulated in the form of an injectable suspension of coated particles having a size distribution that is capable of forming a homogenous and stable (i.e., non-settling) suspension in the injection fluid and that is capable of being injected through a needle. In this regard, the formulations of the present invention may comprise an aqueous medium containing inactive ingredients that has sufficient viscosity to prevent premature gelation of the formulations of the present invention and / or to prevent settling that would result in the suspension not being "homogeneous" and thus risking under- or overdosing of the active ingredient.

[0162] Thus, the formulations can be stored under normal storage conditions and maintain their physical and / or chemical integrity. The phrase "maintain physical and chemical integrity" essentially means chemical stability and physical stability.

[0163] "Chemical stability" includes the ability of the formulations of the present invention to be stored (with or without appropriate pharmaceutical packaging) under normal storage conditions with only slight chemical degradation or decomposition.

[0164] "Physical stability" includes that the formulations of the present invention may be stored (with or without appropriate pharmaceutical packaging) under normal storage conditions with only a small degree of physical transformation (e.g., precipitation as described above) or change in the properties and / or integrity of the coated particles, e.g., the coating itself or the active ingredient (including dissolution, solvation, solid-state phase transitions, etc.).

[0165] Examples of "normal storage conditions" for the formulations of the present invention include temperatures of about -50°C to about +80°C (preferably, about -25°C to about +75°C, e.g., about 50°C) for an extended period of time (i.e., about 12 months or more, e.g., about 6 months), and / or pressures of about 0.1 to about 2 bar (preferably, atmospheric pressure), and / or exposure to about 460 lux of ultraviolet / visible light, and / or relative humidity of about 5 to about 95% (preferably, about 10 to about 40%).

[0166] Under such conditions, the formulations of the invention may be found to be chemically and / or physically degraded / decomposed, optionally to less than about 15%, more preferably less than about 10%, especially less than about 5%.Those skilled in the art will appreciate that the above upper and lower limits of temperature and pressure represent the extremes of normal storage conditions, and that certain combinations of these extremes will not be experienced during normal storage (e.g., a temperature of 50°C and a pressure of 0.1 bar).

[0167] The formulations of the present invention may comprise from about 1% to about 99% by weight, such as from about 10% (such as about 20% by weight, e.g., about 50% by weight) to about 90% by weight of the coated particles, with the remainder being made up of the carrier system and / or other pharma- ceutically acceptable excipients.

[0168] Compositions of the present invention suitable for injection may also include compositions in the form of a liquid, sol, or gel that can be administered via a surgical administration device, such as a needle, catheter, etc., to form a depot preparation.

[0169] In any case, the preparation of suitable formulations can be accomplished non-innovatively by those skilled in the art using routine techniques. Thus, the formulations of the present invention and dosage forms containing same can be formulated with conventional pharmaceutical additives and / or excipients used in the art for the preparation of pharmaceutical formulations, and then incorporated into various types of pharmaceutical formulations and / or dosage forms using standard techniques (see, for example, Lachman et al., 'The Theory and Practice of Industrial Pharmacy', Lea & Febiger, 30, 2003). rd edition (1986), 'Remington: The Science and Practice of Pharmacy', Troy (ed.), University of the Sciences in Philadelphia, 21 st edition (2006), and / or 'Aulton's Pharmaceutics: The Design and Manufacture of Medicines', Aulton and Taylor (eds.), Elsevier, 4 th edition, 2013), and the documents cited therein, the relevant disclosures of all of which are incorporated herein by reference.

[0170] According to a further aspect of the invention there is provided a process for preparing a formulation of the invention comprising mixing a coated particle, e.g. as described herein, together with an aqueous carrier system, e.g. as described herein.

[0171] For parenteral administration, such as subcutaneous and / or intramuscular injection, the formulations of the present invention may be presented in the form of sterile injectable and / or infusible dosage forms that can be administered via a surgical administration device (e.g., a syringe with an injection needle, catheter, etc.) to form a depot preparation.

[0172] Additionally, injectable and / or infusible dosage forms are provided that comprise the formulations of the invention, the formulations being contained within a reservoir that is connected to and / or associated with an injection or infusion means (e.g., a syringe with an injection needle, catheter, etc.).

[0173] Alternatively, the formulations of the invention may be stored prior to loading into a suitable injectable and / or injectable dosage device (e.g., a syringe equipped with a needle for injection) or may be prepared immediately prior to loading into such a dosage device.

[0174] Thus, the sterile injectable and / or infusible dosage forms may comprise a receptacle or reservoir in communication with the injection or infusion means which may be preloaded with the formulation of the invention, which may be loaded prior to use, or may comprise one or more reservoirs in which the formulation of the invention and the coated particles of the aqueous carrier system are separately contained and mixing occurs prior to and / or during injection or infusion.

[0175] therefore, (a) coated particles of a formulation of the invention; (b) a carrier system of the formulation of the present invention; As well as a kit-of-parts comprising coated particles of the formulation of the invention together with instructions to the end user to mix the particles with a carrier system according to the invention.

[0176] There is further provided a pre-loaded injectable and / or infusible dosage form as described above, but modified by including at least two chambers, in one of which the coated particles of the formulation of the invention are disposed, and in the other chamber the aqueous carrier system of the formulation of the invention is disposed, and upon mixing a suspension or otherwise is generated prior to and / or during injection or infusion.

[0177] The formulations of the present invention can be used in human medicine, and are particularly useful in any indication for which a biologically active drug is either approved for use or is otherwise known to be useful.

[0178] The formulations of the present invention are indicated in the curative, palliative, and / or diagnostic treatment, as well as the prophylactic treatment (including preventing and / or arresting the deterioration and / or worsening of a condition) of any condition that the biologically active agent is known to treat.

[0179] Injection of the formulations of the present invention may cause a mild inflammatory response, which may be mitigated by co-administration of a suitable anti-inflammatory agent with the injection.

[0180] Suitable anti-inflammatory agents which may be used in this regard include butylpyrazolidines (e.g., phenylbutazone, mofebutazone, oxyphenbutazone, clofezone, kebuzone and suxibuzone), acetic acid derivatives and related substances (indomethacin, sulindac, tolmetin, zomepirac, diclofenac, alclofenac, bumadizone, etodolac, lonazolac, fentiazac, acemetacin, difenpyramide, oxametacin, proglumetacin, ketorolac, aceclofenac and bufexamac), oxicams (e.g., piroxicam, tenoxicam, droxicam, lornoxicam and meloxicam), propionic acid derivatives (e.g., ibuprofen, naproxen, ketoprofen, fenoprofen, fenbufen, benoxaprofen, suprofen, pirprofen, flurbiprofen, indoprofen, tiaprofenic acid, oxaprozin, ibuproxam, dexibuprofen, flunoxaprofen, alminoprofen, dexketoprofen, vedaprofen, carprofen and tepoxalin), fenamates (e.g., mefenamic acid, tolfenamic acid), , flufenamic acid, meclofenamic acid, and flunixin), coxibs (e.g., celecoxib, rofecoxib, valdecoxib, parecoxib, etoricoxib, lumiracoxib, firocoxib, robenacoxib, mavacoxib, and cimicoxib), other nonsteroidal anti-inflammatory drugs (e.g., nabumetone, niflumic acid, azapropazone, glucosamine, benzydamine, glucosaminoglycan polysulfate, proquazone, orgotein, nimesulide, feprazone, diacerein, morniflumate, tenidap, oxaceprol, condom, leutin sulfate, pentosan polysulfate and aminopropionitrile), corticosteroids (e.g., 11-dehydrocorticosterone, 11-deoxycorticosterone, 11-deoxycortisol, 11-ketoprogesterone, 11β-hydroxypregnenolone, 11β-hydroxyprogesterone, 11β,17α,21-trihydroxypregnenolone, 17α,21-dihydroxypregnenolone, 17α-hydroxypregnenolone, 17α-hydroxyprogesterone, 18-hydroxy-11-deoxycorticosterone,18-hydroxycorticosterone, 18-hydroxyprogesterone, 21-deoxycortisol, 21-deoxycortisone, 21-hydroxypregnenolone (prevedilone), aldosterone, corticosterone (17-deoxycortisol), cortisol (hydrocortisone), cortisone, pregnenolone, progesterone, flugestone (flurogestone), fluorometholone, medrysone (hydroxymethylprogesterone), prevedilone acetate (21-acetoxypregnenolone), chloroprednisone, cloprednol, difluprednate, fludrocortisone, fluocinolone, fluperolone, fluprednisolone, loteprednol , methylprednisolone, prednicarbate, prednisolone, prednisone, tixocortol, triamcinolone, alclometasone, beclomethasone, betamethasone, clobetasol, clobetasone, clocortolone, desoximetasone, dexamethasone, diflorasone, difluocortolone, fluclorolone, flumethasone, fluocortin, fluocortolone, fluprednidene, fluticasone, fluticasone furoate, halometasone, meprednisone, mometasone, mometasone furoate, paramethasone, prednylidene, rimexolone, urobetasol (halobetasol), amcinonide, budesonide, ciclesonide, deflazacort, desonide, formocortal Fluclorone acetonide (flucloronide), fludroxycortide (flurandrenolon, flurandrenolide), flunisolide, fluocinolone acetonide, fluocinonide, halcinonide and triamcinolone acetonide), quinolines (e.g., oxycinchophen), gold preparations (e.g., sodium gold thiomalate, sodium aurothiosulfate, auranofin, aurothioglucose and aurothioprole), Penicillamine and similar agents (such as bucillamine), and antihistamines (acrivastine, alimemazine, antazoline, astemizole, azatadine, azelastine, bamipine, bilastine, bromdiphenhydramine, brompheniramine, buclizine, cetirizine, cinnarizine, cyclizine, cyproheptadine, deptropine, desloratadine, dexbrompheniramine, dexchlorpheniramine,Dienylpyraline, dimenhydrinate, dimethindene, doxylamine, ebastine, epinastine, phenindamine, pheniramine, fexofenadine, histapirodine, hydroxyethylpromethazine, isothipendyl, carbinoxamine, ketotifen, quifenadine, clemastine, chlorcyclizine, chlorphenamine, chlorphenoxamine, chloropyramine, levocetirizine, loratadine , mebhydroline, mequitazine, meclozine, mepyramine, methapyrilene, methdilazine, mizolastine, oxatomide, oxomemazine, pimethixene, promethazine, pyrobutamine, rupatadine, sequifenadine, talastine, thenarizine, terfenadine, thiazinam, thiethylperazine, thonzylamine, trimethobenzamide, tripelennamine, triprolidine and tritocubaline). Combinations of any one or more of the foregoing anti-inflammatory agents may be used.

[0181] Preferred anti-inflammatory agents include nonsteroidal anti-inflammatory drugs such as diclofenac, ketoprofen, meloxicam, aceclofenac, flurbiprofen, parecoxib, ketolac tromethamine, or indomethacin.

[0182] The subject may receive (or may already receive) one or more of the aforementioned anti-inflammatory agents separate from the formulation of the present invention, meaning that they receive a prescribed dose of one or more of these anti-inflammatory agents prior to, in addition to, and / or following treatment with the formulation of the present invention.

[0183] When a biologically active agent is "combined" with such an anti-inflammatory agent, the active ingredients may be administered together in the same formulation or may be administered separately (simultaneously or sequentially) in different formulations (hereinafter referred to as a "combination product").

[0184] Such combination products provide for administration of a biologically active agent in conjunction with an anti-inflammatory agent and therefore may be presented as separate formulations, where at least one of the formulations is a formulation of the invention and at least one includes an anti-inflammatory agent in a separate formulation, or may be presented (i.e., formulated) as a combined preparation (i.e., may be presented as a single formulation including a biologically active agent and an anti-inflammatory agent).

[0185] In this regard, the anti-inflammatory agent may be presented together with the biologically active agent in an appropriate dose in one or more of the cores forming part of the formulation of the invention described above, or may be formulated using the same or similar processes for coating as those described above for the biologically active agent, which may allow release of the anti-inflammatory agent over the same time scale or over a different time scale.

[0186] Thus, there is further provided a pharmaceutical formulation of the invention which further comprises an anti-inflammatory agent.

[0187] In such formulations of the invention, the anti-inflammatory agent is: (1) formulating within the solid core of a formulation of the present invention (which formulation is hereinafter referred to as a "combination core preparation") with a bioactive agent; (2) It may be included by dissolving and / or suspending it in the aqueous carrier system of the formulation of the present invention (which formulation is hereinafter referred to as the "combination preparation").

[0188] In embodiment (2) above, the anti-inflammatory agent may be presented in the formulation of the invention in any form separate from the biologically active agent-containing core. This may be achieved, for example, by dissolving or suspending the active ingredient directly in the aqueous medium of the formulation of the invention, or by presenting it in a form whose release, like the biologically active agent, may be controlled after injection.

[0189] The latter option may be achieved, for example, by providing the anti-inflammatory agent in the form of additional particles suspended in the aqueous carrier system of the formulation of the invention, the additional particles having an average diameter by weight, number, or volume that is from about 10 nm to about 700 μm, and comprising a core that contains the anti-inflammatory agent, the core being at least partially coated with one or more coating materials as described above (this formulation is hereinafter referred to as a "combination suspension").

[0190] It may also be in the form of a kit of parts, comprising the following components: (A) a pharmaceutical formulation of the present invention; (B) a pharmaceutical formulation comprising an anti-inflammatory agent, There is provided a pharmaceutical formulation of the invention in the form of a kit of parts, in which these components (A) and (B) are each provided in a form suitable for administration in conjunction with the other.

[0191] Component (B) of the above proposed kit-of-parts may differ in its chemical composition and / or physical form from component (A) (i.e., the formulation of the present invention), but it may also be essentially in the same or at least similar form as the biologically active agent-containing formulation of the present invention, i.e., in the form of a plurality of particles suspended in a (e.g., aqueous) carrier system, said particles being: (a) have an average diameter based on weight, number, or volume that is from about 10 nm to about 700 μm; (b) comprises a solid core containing the anti-inflammatory agent, which is at least partially coated with one or more coatings of (eg, inorganic) materials.

[0192] In addition, in such preferred kit-of-parts, and combination suspensions presented under embodiment (2) above, the coated cores comprising the anti-inflammatory agent may differ in terms of their chemical composition and / or physical form, but the coating of inorganic material used is preferably the same or similar to the coating used in the biologically active agent-containing formulation of the present invention, meaning that the anti-inflammatory agent is coated with one or more inorganic coatings as described herein, e.g., one or more inorganic coating materials comprising one or more metal-containing or metalloid-containing compounds, e.g., metals, or metalloids, oxides, e.g., iron oxide, titanium dioxide, zinc sulfide, more preferably zinc oxide, silicon dioxide and / or aluminum oxide, which coating materials essentially constitute (e.g., on an individual or collective basis) (e.g., greater than about 80%, e.g., about 90%, e.g., about 95%, e.g., about 98%) of such oxides, and more specifically, the inorganic coating is: (i) zinc oxide; (ii) In admixture with one or more other metal and / or metalloid oxides. The atomic ratio ((i):(ii)) is between at least about 1:6 and at most about 6:1.

[0193] Preferably, the atomic ratio ((i):(ii)) is between at least about 1:1 and at most about 6:1.

[0194] In any event, for the avoidance of doubt, all aspects, including preferred aspects, of the biologically active agent-containing formulations of the present invention disclosed and / or claimed herein are equally applicable as aspects and / or preferences of the coated cores comprising one or more of the anti-inflammatory agents described above, and for the avoidance of doubt, such aspects, preferences and features, either singly or in combination, are hereby incorporated by reference to these aspects of the present invention.

[0195] Thus, all combination products, including the above combination core preparations, combination suspensions and kits of parts, may be used in human medicine, particularly for any indication where a biologically active agent is approved for use or is otherwise known to be useful.

[0196] According to a further aspect of the invention there is provided a method of making a kit-of-parts as defined above, the method comprising bringing component (A), as defined above, into association with component (B), as defined above, thus rendering the two components suitable for administration in combination with one another.

[0197] By "associating" the two components with each other, components (A) and (B) of the kit are: (i) may be provided as separate formulations (i.e., independent of each other) and then combined for use in conjunction with each other in a combination therapy; or (ii) They may be packaged and presented together as separate components of a "combination pack" for use in conjunction with each other in combination therapy.

[0198] Thus, a kit-of-parts as defined herein, wherein components (A) and (B) are packaged and presented together as separate components of a combination pack, for use with each other in a combination therapy, and: (I) one of components (A) and (B) as defined herein; (II) instructions for using the component in conjunction with the other of the two components.

[0199] As mentioned above, the kit-of-parts described herein may include two or more formulations containing suitable amounts / doses of a biologically active agent and / or two or more formulations containing suitable amounts / doses of an anti-inflammatory agent to provide for repeated administration, as mentioned above.

[0200] In this regard, with respect to the kit-of-parts described herein, "administration in conjunction with" includes that components (A) and (B) of the kit are administered sequentially, separately and / or simultaneously over the course of treatment of a condition.

[0201] Thus, the term "in conjunction with" includes that one or the other of the two formulations may be administered (optionally repeatedly) before, after, and / or simultaneously with the administration of the other component. As used in this context, the terms "co-administered" and "administered simultaneously with" include individual doses of the biologically active agent and the anti-inflammatory agent being administered within 48 hours (e.g., 24 hours) of each other.

[0202] With respect to any of the above combination products according to the invention, each formulation is administered (or, in the case of a kit-of-parts, the two components are administered together, optionally repeatedly), in a manner that may enable a beneficial effect for the subject over the course of treatment of the condition, i.e., a greater extent, than if a formulation comprising the biologically active agent alone (e.g., a formulation of the invention) were administered over the same course of treatment (e.g., repeatedly as described herein) in the absence of the other component.

[0203] The determination of whether a combination product provides a greater beneficial effect with respect to treatment and over the course of treatment depends on the condition being treated and / or its severity, but can be routinely accomplished by one of ordinary skill in the art.

[0204] The physician may then administer one or more of the following: ● The above kit-of-parts components (B), Combination core preparations, Concomitant preparations, and / or Combined suspension As above, either of these may include an anti-inflammatory agent as previously described.

[0205] The amount of anti-inflammatory agent that may be used in a combination product according to the present invention should be sufficient to exert its pharmacological effect.

[0206] Therefore, the dose of such anti-inflammatory components that can be administered to a patient must be sufficient to affect a therapeutic response over a reasonable and / or relevant time frame.Those skilled in the art recognize that the selection of the exact dose and composition and the most suitable delivery regimen is influenced not only by the nature of the anti-inflammatory agent, but also by, among others, the pharmacological properties of the formulation, the route of administration, the nature and severity of the condition being treated, the physical condition and mental acuity of the recipient, and the age, condition, weight, sex, and response of the patient being treated, the stage / severity of the disease, and genetic differences between patients.

[0207] Because administration of the formulations of the present invention can be continuous or intermittent (eg, by bolus injection), the dosage of such anti-inflammatory components can also be determined by the timing and frequency of administration.

[0208] In any event, a physician or other skilled artisan can routinely determine the actual dosage of any particular additional active ingredient that will be most suitable for an individual patient, and dosages of such relevant additional active ingredients are known in the art and are set forth in the medical literature (Martindale-The Complete Drug Reference, 38 U.S.C. 5,311,133; 5,311,133; and 5,311,133). th Edition, Pharmaceutical Press, London (2014) and documents referenced therein, the relevant disclosures of all of which are incorporated herein by reference.

[0209] The use of the formulations of the present invention may be advantageous in reducing any burst effect (e.g., concentration maxima immediately following administration) as discussed above and / or in reducing the C max By lowering the dissolution rate, it is possible to control the dissolution rate of the biologically active drug and affect the pharmacokinetic profile.

[0210] The formulations of the present invention may also provide release and / or pharmacokinetic profiles that increase the length of release of the biologically active drug from the formulation.

[0211] These factors not only reduce the frequency or frequency with which the formulation needs to be administered to the subject, but also allow the subject more time as an outpatient, thus improving their quality of life.

[0212] The formulations of the invention also have the advantage that by controlling the release of the active ingredient at a steady rate over an extended period of time, for example, they provide a lower daily exposure to cytotoxic drugs, which is expected to reduce undesirable side effects.

[0213] The formulations and processes described herein may have advantages for the physician and / or patient in treating the relevant condition, such as being more convenient, more effective, less toxic, having a broader spectrum of activity, more potent, causing fewer side effects, and having other useful pharmacological properties, than any similar treatments known in the prior art.

[0214] Whenever the word "about" is used herein, for example in the context of an amount (e.g., number, concentration, dimension (size and / or weight), dose, duration, pharmacokinetic parameter, etc.), relative amount (percentage, weight ratio, size ratio, atomic ratio, aspect ratio, proportion, multiple or fraction, etc.), relative humidity, lux, temperature or pressure, it will be understood that such variables are approximate and thus may vary from the numerical values ​​specified herein by ±15%, such as ±10%, for example ±5%, preferably ±2% (e.g. ±1%). This is true even when such numerical values ​​are expressed as percentages in the first place (e.g., "about 15%" may mean ±15% of the numerical value 10, which is anywhere from 8.5% to 11.5%).

[0215] The invention is illustrated, but in no way limited, by the following examples with reference to the figures, in which Figure 1 shows the absorbance measured at 450 nm in a UV-VIS plate reader after ELISA on coated and uncoated spray-dried microparticles containing monoclonal antibody ATH3G10. EXAMPLES

[0216] Example 1 Mixed oxide coated Anakin particulate I. A microparticle sample of anakinra was prepared by spray drying with trehalose to give a mean particle diameter of 5 μm as determined by laser diffraction.

[0217] The powder is loaded into an ALD reactor (Picosun, SUNALE™ R-series, Espoo, Finland) and 24 ALD cycles are run at a reactor temperature of 50 °C. The coating sequence consists of three ALD cycles with diethylzinc and water as precursors for three ALD cycles, followed by one cycle of trimethylaluminum and water, repeated six times to form a mixed oxide layer with a zinc:aluminum atomic ratio of 3:1. The first layer is expected to be approximately 5 nm thick (estimated from the number of ALD cycles).

[0218] The powder was removed from the reactor and deagglomerated by forcing it through a polymeric sieve with a mesh size of 20 μm using a sonic sifter.

[0219] The resulting deagglomerated powder was reloaded into the ALD reactor and subjected to 24 more ALD cycles as before to form a second layer of the mixed oxide in the ratios described above, extracted from the reactor, deagglomerated by means of sonic sieving as above, reloaded to form a third layer, deagglomerated, and then reloaded to form a final, fourth layer.

[0220] To determine the drug loading (i.e. w / w% of anakinra in the powder), it is planned to determine the drug loading using methods known in the art. The nanoshell coating was dissolved in 2 M phosphoric acid in DMSO, and the slurry was then diluted with DMSO before being filtered (0.2 μm RC, Lab Logistics Group, Germany) and further analyzed by HPLC (n=2).

[0221] Example 2 Mixed oxide coated Anakin microparticles II The same procedure as described in Example 1 is carried out to produce microparticles coated with a mixed oxide coating containing an atomic ratio of zinc:aluminum of 2:1.

[0222] The coating sequence was two ALD cycles using diethylzinc and water as precursors, followed by one cycle of trimethylaluminum and water, repeatedly removing the coated powder from the reactor 10 times, deagglomerating, reloading, and repeating the same coating sequence, removing, and deagglomerating until a total of four sets of 30 cycles were provided.

[0223] Example 3 Aluminum oxide coated Anakin microparticles The same particulates coated with the mixed oxide coating as described in Example 1 are coated with a pure aluminum oxide coating. Thirty ALD cycles are performed before the coated powder is removed from the reactor and deagglomerated as described in Example 1. The resulting deagglomerated powder is reloaded into the ALD reactor and subjected to an additional 30 ALD cycles, followed by extraction, deagglomeration, reloading, repeating the same coating sequence, removing, and deagglomerating until a total of four sets of 30 cycles are provided.

[0224] Example 4 Zinc oxide coated Anakin microparticles The same particulates coated with the mixed oxide coating as described in Example 1 are coated with a pure zinc oxide coating. Thirty ALD cycles are performed before the coated powder is removed from the reactor and deagglomerated as described in Example 1. The resulting deagglomerated powder is reloaded into the ALD reactor and subjected to an additional 30 ALD cycles, followed by extraction, deagglomeration, reloading, repeating the same coating sequence, removing, and deagglomerating until a total of four sets of 30 cycles are provided.

[0225] Example 5 Formulation I of the Invention The same microparticles coated with the oxide coating described in Examples 1, 2, 3 and 4 are suspended in a commercially available aqueous vehicle, Hyonate® vet (Boehringer Ingelheim Animal Health, France), a veterinary drug used for injections of animals such as horses, which contains a sterile, isotonic, phosphate buffer solution of 10 mg / mL sodium hyaluronate, pH 7.4.

[0226] The concentration of anakin in the formulation was 10 mg / mL, which corresponds to 10 mg / kg body weight in Sprague-Dawley rats.

[0227] Example 6 Formulation II of the Invention A suspension of coated microparticles of anakinra (prepared according to the process described in Example 4 above) is suspended in an aqueous vehicle containing 0.1% (w / w) Polysorbate 20, 0.25% (w / w) sodium carboxymethylcellulose in phosphate buffered saline (pH 7.4).

[0228] The concentration of anakin in the formulation was 10 mg / mL, which corresponds to 10 mg / kg body weight in Sprague-Dawley rats.

[0229] Example 7 Formulation III of the Invention A suspension of anakinra uncoated microparticles (the same type of uncoated microparticles used for the coated microparticles of Examples 1-4) is suspended in commercially available aqueous BiHyonate® vet to a concentration of azacytidine in the formulation of 10 mg / mL, corresponding to 10 mg / kg body weight in Sprague-Dawley rats.

[0230] Example 8 In vivo rat study I Thirty-six male Sprague Dawley rats are provided, for example, by Charles River (UK). The animals are randomly divided into groups of 6 animals.

[0231] The hair in the intended administration area is clipped prior to injection and the injection site is marked. As shown in Table 1 below, the suspensions described in Example 5 (Group 1 suspension of particles described in Example 1, Group 2 suspension of particles described in Example 2, Group 3 suspension of particles described in Example 3, and Group 4 suspension of particles described in Example 4), Example 6 (Group 5), and Example 7 (Group 6) are drawn into a 1 mL BD syringe and a single subcutaneous injection (approximately 0.3 mL) is administered through a 23G needle (BD Microlance) into the flank of each rat. Administration occurs within 30 minutes of preparation of the formulation. [Table 1]

[0232] Blood samples (approximately 0.2 mL) are collected into K2EDTA (dipotassium ethylenediaminetetraacetate) tubes from the tail vein at the following time points: 0.5, 1, 3, 6, 12, 24, 48, 72, 120, 168, 240, and 336 hours post-dose. Actual sampling times are recorded. As soon as practically possible after blood sampling, plasma is separated by centrifugation (1500 g, 10 min at 4° C.) and stored at −80° C. until analysis is performed.

[0233] Upon completion of the study, all plasma samples are shipped for analysis deep frozen on dry ice. Animals are sacrificed on the final day of the study.

[0234] Plasma concentrations of anakinra are determined using HPLC-MS / MS.

[0235] The relative bioavailability of the formulations is expected to be similar to that of the uncoated biologically active drug.

[0236] Pharmacokinetic analysis of anakinra in plasma was performed following a standard non-compartmental approach using Microsoft Excel for Mac (16.43, Microsoft, Redmond, Washington, USA). Maximum concentration, C max , and the associated time, t max is the coordinate of the maximum concentration in the time course. t last is the time to the last detectable concentration. last ) is calculated using the linear trapezoidal rule.

[0237] result Dose-normalized plasma concentrations of anakinra after a single subcutaneous administration of various formulations are presented. Plasma pharmacokinetic parameters are also presented as mean values ​​for groups of six rats (standard deviations are shown in brackets). • Doses are expressed in mg / kg body weight in rats. ● "t max " is the time to peak concentration expressed in hours. ● "C max " is the maximum concentration found in the assay expressed in μg / mL. ● "t last " is the time of last detectable concentration expressed in hours. ● "t 1 / 2,z " is the terminal half-life expressed in hours. ● "AUC ∞ " is the area under the concentration versus time curve to infinity expressed in μg*h / mL. • "F" is the relative bioavailability expressed as a percentage. ● "C max / D" is the maximum concentration normalized to 1 mg / kg expressed in μg / mL / mg / kg body weight in rats. ● "AUC last " / D" is the area under the blood concentration versus time curve to the last detectable concentration normalized to 1 mg / kg expressed in μg*h / mL / mg / kg body weight of the rat. ● "AUC ∞ / D" is the area under the concentration versus time curve to infinity normalized to 1 mg / kg, expressed in μg*h / mL / mg / kg body weight of rats. ● “Fr. Rel. 0-12h " is the fraction released during the first 12 hours of the area under the concentration versus time curve to infinity expressed as a percentage.

[0238] The following order is suspected in terms of drug release (fastest to slowest): group 6 group 4 group 1 group 2 group 3

[0239] The formulations in Group 5, ie, Example 6, are more likely to form gels that would be difficult to administer.

[0240] Example 9 Formulation IV of the Invention The same microparticles coated with the oxide coating described in Examples 1, 2, 3, and 4 are suspended in the commercially available aqueous vehicle Hyonate® vet (Boehringer Ingelheim Animal Health, France), a veterinary drug used for injections of animals such as horses, which contains a sterile, isotonic, phosphate buffer solution of 200 mg / mL sodium hyaluronate (pH 7.4).

[0241] The concentration of anakinra in the formulation was 200 mg / mL, which corresponds to 200 mg / kg body weight in Sprague-Dawley rats.

[0242] Example 10 Formulation V of the Invention A suspension of coated microparticles of anakinra (prepared according to the process described in Example 4 above) is suspended in an aqueous vehicle containing 0.1% (w / w) Polysorbate 20, 0.25% (w / w) sodium carboxymethylcellulose in phosphate buffered saline (pH 7.4).

[0243] The concentration of anakinra in the formulation was 200 mg / mL, which corresponds to 200 mg / kg body weight in Sprague-Dawley rats.

[0244] Example 11 Formulation VI of the Invention A suspension of anakinra uncoated microparticles (the same type of uncoated microparticles used for the coated microparticles of Examples 1-4) is suspended in the commercially available aqueous vehicle Hyonate® vet to a concentration of azacytidine in the formulation of 10 mg / mL, equivalent to 200 mg / kg body weight in Sprague-Dawley rats.

[0245] Example 12 In vivo rat study II Thirty-six male Sprague Dawley rats are provided, for example, by Charles River (UK). The animals are randomly divided into groups of 6 animals.

[0246] The hair in the intended administration area is clipped prior to injection and the injection site is marked. As shown in Table 2 below, the suspensions described in Example 9 (Group 1 suspension of particles described in Example 1, Group 2 suspension of particles described in Example 2, Group 3 suspension of particles described in Example 3, and Group 4 suspension of particles described in Example 4), Example 10 (Group 5), and Example 11 (Group 6) are drawn into a 1 mL BD syringe and a single subcutaneous injection (approximately 0.3 mL) is administered through a 23G needle (BD Microlance) into the flank of each rat. Administration is performed within 30 minutes after preparation of the formulation. [Table 2]

[0247] Blood samples (approximately 0.2 mL) are collected into K2EDTA (dipotassium ethylenediaminetetraacetate) tubes from the tail vein at the following time points: 0.5, 1, 3, 6, 12, 24, 48, 72, 120, 168, 240, and 336 hours post-dose. Actual sampling times are recorded. As soon as practically possible after blood sampling, plasma is separated by centrifugation (1500 g, 10 min at 4° C.) and stored at −80° C. until analysis is performed.

[0248] Upon completion of the study, all plasma samples are shipped for analysis deep frozen on dry ice. Animals are sacrificed on the final day of the study.

[0249] Plasma concentrations of anakinra are determined using HPLC-MS / MS.

[0250] The relative bioavailability of the formulations is expected to be similar to that of the uncoated biologically active drug.

[0251] Pharmacokinetic analysis of anakinra in plasma was performed following a standard non-compartmental approach using Microsoft Excel for Mac (16.43, Microsoft, Redmond, Washington, USA). Maximum concentration, C max , and the associated time, t max is the coordinate of the maximum concentration in the time course. t last is the time to the last detectable concentration. last ) is calculated using the linear trapezoidal rule.

[0252] result Dose-normalized plasma concentrations of anakinra after a single subcutaneous administration of various formulations are presented. Plasma pharmacokinetic parameters are also presented as mean values ​​for groups of six rats (standard deviations are shown in brackets). • Doses are expressed in mg / kg body weight in rats. ● "t max " is the time to peak concentration expressed in hours. ● "C max " is the maximum concentration found in the assay expressed in μg / mL. ● "t last " is the time of last detectable concentration expressed in hours. ● "t 1 / 2,z " is the terminal half-life expressed in hours. ● "AUC ∞ " is the area under the concentration versus time curve to infinity expressed in μg*h / mL. • "F" is the relative bioavailability expressed as a percentage. ● "C max / D" is the maximum concentration normalized to 1 mg / kg expressed in μg / mL / mg / kg body weight in rats. ● "AUC last " / D" is the area under the blood concentration versus time curve to the last detectable concentration normalized to 1 mg / kg expressed in μg*h / mL / mg / kg body weight of the rat. ● "AUC ∞ / D" is the area under the concentration versus time curve to infinity normalized to 1 mg / kg, expressed in μg*h / mL / mg / kg body weight of rats. ● “Fr. Rel. 0-12h " is the fraction released during the first 12 hours of the area under the concentration versus time curve to infinity expressed as a percentage.

[0253] The following order is suspected in terms of drug release (fastest to slowest): group 6 group 4 group 1 group 2 group 3

[0254] The formulations in Group 5, ie, Example 10, are more likely to form gels that would be difficult to administer.

[0255] Example 13 Coated Monoclonal Antibody ATH3G10 Clinical Trial Material A solution of fully human IgG1 monoclonal antibody with specific affinity for phosphocholine (PC mAb) dissolved at 20 mg / mL in an aqueous solution of 125 mM sodium chloride, 100 mM glycine, and 25 mM sodium acetate (pH 5.5) was obtained from Athera Biotechnologies AB (Stockholm, Sweden). The production of this specific monoclonal antibody is described in de Vries et al. (2021), J. Intern. Med. 290(1), pp. 141-156, and general procedures for producing anti-phosphorylcholine antibodies are described in the art, e.g., U.S. Pat. No. 5,455,032. In summary, the described antibodies can be produced by synthesizing DNA encoding the antibody sequence and cloning it into a plasmid for transfection into 293T cells. The cells then transiently produce the antibody, which can be purified, e.g., using a protein-A sepharose column.

[0256] For large-scale production, stable cell lines have been developed. Gene-optimized DNA sequences encoding the signal peptide and coding regions of the variable regions of the heavy (VH) and light (VL) chains of the X19-A05 antibody were generated by Geneart AG (Regensburg, Germany). The DNA sequence encoding the VH was cloned into a vector containing a gene-optimized cDNA sequence encoding the human IgG1za constant region, except for the codon encoding the C-terminal lysine residue. The DNA sequence encoding the VL was cloned into a vector containing a gene-optimized cDNA sequence encoding the human kappa constant region. CHOK1SV host cells were transfected with a single vector encoding both the complete heavy and light chain genes to generate a stable GS-CHO transfection pool expressing PC mAb. The cell line 3G10 was selected for cGMP manufacturing of the PC-mAb antibody, designated ATH3G10. Vector construction, cell line generation, and antibody production were performed by Lonza Biologics plc (Slough, UK).

[0257] ATH3G10 clinical trial material was reformulated by dialysis through a 20 kDa MWCO membrane. The dialysis process was carried out against 1:10 volume of water for injection for 3 hours, after which the dialysate was replaced with 1:10 volume of buffer containing 5 mg / mL DL-histidine in water, adjusted to pH 6.0 using glacial acetic acid. After 3 hours, the dialysate was replaced with 1:10 volume of buffer containing 3.3 mg / mL trehalose and 4.5 mg / mL DL-histidine in water, adjusted to pH 6.0 using glacial acetic acid, and dialysis was continued for another 12 hours. Following the dialysis process, titrator test strips (30-600 mg / L Cl) were used to measure the pH of the dialysate. - ) was used to measure chloride concentrations. After dialysis, the composition of the retentate was 0.8–1.6% trehalose, 3.4–3.9% DL-histidine, 0.4–0.5% glycine, 0.3–0.5% sodium chloride, and 0.1% sodium acetate in water. Additional trehalose was added to obtain a final concentration of 3.7%–4.1% trehalose in the retentate. The retentate was then spray-dried using a Büchi B-290 Mini (Essen, Germany) with an inlet temperature of 110 °C, an aspirator speed of 100%, a pump speed of 4.6 mL / min, and a nebulizing gas (N2) volumetric flow rate of 600–700 L / h, resulting in an outlet temperature of 65–67 °C. The spray-drying process took 45 min and yielded >90%. A fine white powder of amorphous particles with toroidal and combined toroidal morphology was obtained, with an average particle size of 3-5 μm as determined by scanning electron microscopy.

[0258] Microparticle samples of ATH3G10, a fully human IgG1 monoclonal antibody with specific affinity for phosphocholine (Athera Biotechnologies, Stockholm, Sweden), were prepared by spray drying with trehalose and DL-histidine to yield particles with toroidal and combined toroidal morphology and a mean particle diameter of 5 μm as determined by scanning electron microscopy.

[0259] The powder was loaded into an ALD reactor (Picosun, SUNALE™ R-series, Espoo, Finland) and 20 ALD cycles were carried out at a reactor temperature of 30° C. The coating sequence was trimethylaluminum and water as precursors, which were pulsed into the reactor by means of a stopped-flow process. This means that each precursor was left in the reaction chamber without active pumping so that all surfaces could be coated. The particulates were subjected to 20 ALD cycles to create a first layer of aluminum oxide.

[0260] The ALD reactor included a reaction chamber loaded with particulates. The ALD reactor further included precursor bottles separately containing each precursor, with each precursor bottle being coupled to the reaction chamber via a valve. The ALD reactor also included a pump and associated piping for pumping an inert gas, such as nitrogen, through the reaction chamber, with the pump also being coupled to the reaction chamber via a valve.

[0261] An ALD cycle is performed as follows, where steps a through d represent the first cycle, and subsequent cycles begin with step a, as specified in step e. a. Reagent Pulse 1: i. The valve in the piping between the pump and the ALD reactor was closed. ii. The valve on the water precursor bottle was opened for 0.5 seconds to fill the reaction chamber with evaporated water. iii. The valve to the water precursor bottle was closed and the chamber was allowed to rest for 30 seconds (soak time) before reopening the pump valve to ensure that the water molecules were adsorbed onto the surface of the drug particles, presenting hydroxyl groups to the exterior or particles. iv. The reactor was then pumped for 9 seconds. v. The above steps i to iv were repeated 20 times. b. Purge pulse: The chamber was purged with a continuous flow of nitrogen to remove gaseous water and organic gases. c. Reagent Pulse 2: i. The valve in the piping between the pump and the ALD reactor was closed. ii. The valve on the trimethylaluminum precursor bottle was opened for 0.5 seconds to charge the vaporized metal-containing precursor into the reaction chamber. iii. The valve to the precursor bottle was closed and the chamber was allowed to rest for 30 seconds (soak time) before reopening to the pump to ensure that the metal-containing precursor vapor reacted with the hydroxyl groups on the surface of the drug particles. iv. The reactor was then pumped for 9 seconds. v. The above steps i to iv were repeated 20 times. d. Purge pulse: The chamber was purged with a continuous flow of nitrogen to remove unreacted reagents and organic gases. e. The cycle from steps a to d was repeated 20 times.

[0262] 2. The powder was then removed from the reactor and deagglomerated through a sieve with a mesh size of 20 μm by a sonic sifter (Tsutsui Sonic Agitated Sifting Machine SW-20AT).

[0263] 3. The resulting deagglomerated powder was reloaded into the ALD reactor and step 1 was repeated once to form a second layer of aluminum oxide.

[0264] analysis Drug loading (i.e., w / w% of ATH3G10 in the powder) was estimated through sodium chloride concentration from buffer displacement (via dialysis) as measured by chloride titrator strips. Combined with the estimated yield after spray drying, the drug loading in the uncoated particles was estimated to be approximately 69% ATH3G10, with the remainder including trehalose, histidine, water, glycine, sodium chloride, and sodium acetate (in descending order of mass fraction). Drug loading after coating was estimated gravimetrically by powder mass before and after coating. Drug loading of the coated microparticles was estimated to be approximately 44%.

[0265] Example 14 Coated monoclonal antibody formulations The microparticles of Example 13 above were suspended in a vehicle containing 0.05% (w / v) polysorbate 20, 20 mM monopotassium phosphate, 0.8% (w / v) sodium chloride, 0.02% (w / v) potassium chloride, and water for injection, pH 7.2. The tested concentrations of ATH3G10 in the formulation were 132 and 200 mg / mL.

[0266] The formulations were tested for injectability using the TA.XTplus Texture Analyser and Exponent software, which was used to measure the force required to extend the piston.

[0267] In the method, the instrument piston was connected to a syringe piston (1 mL, 3-part luer lock, polypropylene housing) with a 25-gauge hypodermic needle (25 mm long) filled with 1 mL of formulation. For reference, spray-dried (uncoated) microparticles were dissolved in vehicle to the same ATH3G10 concentration and tested using this method. The vehicle of the reference solution contained 125 mM sodium chloride, 100 mM glycine, 25 mM sodium acetate, and 10 mM phosphate in water at pH 5.8. During the test, the piston was extended at 10 mm / s until a threshold force of 50 N was measured, corresponding to the complete distribution of the syringe contents into air. The threshold of 15 N was considered the maximum tolerable force required for injection.

[0268] result Injectability testing showed that higher forces were required to inject the coated microparticle suspension compared to the corresponding ATH3G10 solution. However, the average force throughout the injection of the coated microparticle suspension was acceptable, corresponding to 132 mg / mL ATH3G10 through a 25-gauge hypodermic needle. The average force recorded in this condition was 5.03±0.16 N, while the corresponding average force recorded for the solution formulation at 132 mg / mL ATH3G10 was 3.97±0.04 N.

[0269] Thus, at approximately 43% drug loading (which may be considered a worst-case scenario due to the high specific surface area of ​​the particles produced by the spray drying process), a highly concentrated dose can still be injected through a 25-gauge hypodermic needle.

[0270] These conditions are expected to be suitable for subcutaneous administration without the patient experiencing significant pain during injection. The injectability of the coated microparticle suspension formulation was not improved but was similar compared to the equivalent solution formulation. However, the mixed metal oxide coating provides a sustained release property for the suspension formulation, which may contribute to reducing the required dosing frequency. Furthermore, the mixed metal oxide coating has been shown to function as an efficient moisture barrier, which can extend the shelf life of the product.

[0271] Example 15 Affinity Testing To determine whether the affinity of ATH3G10 for its antigen is retained after being subjected to ALD coating, the coated ATH3G10 microparticles of Example 13 were tested using an indirect enzyme-linked immunosorbent assay (ELISA) and compared to uncoated ATH3G10.

[0272] The coated material (9 mg) was dispersed in 5 mL of ethylenediaminetetraacetic acid solution (0.02%, in 0.5 mM Dulbecco's phosphate-buffered saline). After 4 h of overhead rotation at 75 rpm, the dispersion was filtered through regenerated cellulose (pore size 0.2 μm) and diluted to 1.5 μg / mL in phosphate-buffered saline containing 0.05% (w / v) polysorbate 20. 96-well microtiter plates precoated with phosphocholine (CVDefine ELISA kit) were used for the assay.

[0273] Non-specific binding was blocked by incubating the plates for 30 min with wells filled with blocking buffer containing 2% (w / w) bovine serum albumin in dilution medium. After discarding the blocking buffer, the diluted solutions were added to the wells. The contents of the wells were then discarded and the wells were washed with dilution medium according to standard protocols.

[0274] Following this, the solution was added to the wells containing a secondary antibody (mouse anti-human IgG1) conjugated with horseradish peroxidase. After discarding the well contents, the wells were washed again, followed by the addition of a substrate solution containing 3,3',5,5'-tetramethylbenzidine. After 15 minutes, the reaction was stopped by adding a sulfuric acid solution (0.5 M) and the absorbance of each well was measured using a Tecan Sunrise UV-VIS plate reader equipped with a 450 nm optical filter.

[0275] result Measurement of absorbance in the wells after ELISA showed that the coated ATH3G10 obtained as described in Example 13 retained its affinity compared to the uncoated antibody. The results are shown in Figure 1 (*p<0.05 calculated using unpaired two-tailed t-test).

[0276] Significance testing using an unpaired two-tailed t-test showed a small but significant (p=0.02) increase in affinity after coating (NB this increase is likely an artifact due to slight uncertainty in the drug loading of the coated particles). However, it was concluded that the process of coating the microparticles as described in Example 13 above does not adversely affect ATH3G10 in terms of its affinity for its antigen.

Claims

**Claim 1** A pharmaceutical formulation comprising a plurality of particles suspended in a carrier system, wherein the particles: (a) have an average diameter, based on weight, number, or volume, of from about 10 nm to about 700 μm, (b) comprise a solid core comprising a biologically active drug at least partially coated by a coating of an inorganic material, The formulation comprising a concentration of the biologically active drug of at least 50 mg / mL. A pharmaceutical formulation. **Claim 2** The pharmaceutical formulation according to claim 1, wherein the concentration of the biologically active drug is at least 250 mg / mL. **Claim 3** The pharmaceutical formulation according to claim 1 or 2, wherein the concentration of the biologically active drug is at least 450 mg / mL. **Claim 4** The pharmaceutical formulation according to claim 1 or 2, wherein the biologically active drug is selected from immunoglobulins, monoclonal antibodies, antibody mimetics, cytokines, or antagonists or agonists of cytokine receptors. **Claim 5** The biologically active drug is an immunoglobulin and is selected from immunoglobulins, normal human, for extravascular administration (J06BA01), immunoglobulins, normal human, for intravascular administration (J06BA02), anti-D (rh) immunoglobulin (J06BB01), tetanus immunoglobulin (J06BB02), human growth hormone, varicella / zoster immunoglobulin (J06BB03), hepatitis B immunoglobulin (j06BB04), rabies immunoglobulin (J06BB05), rubella immunoglobulin (J06BB06), vaccinia immunoglobulin (J06BB07), staphylococcus immunoglobulin (J06BB08), cytomegalovirus immunoglobulin (J06BB09), diphtheria immunoglobulin (J06BB10), hepatitis A immunoglobulin (J06BB11), encephalitis, tick-borne immunoglobulin (J06BB12), pertussis immunoglobulin (J06BB13), measles immunoglobulin (J06BB14), mumps immunoglobulin (J06BB15), palivizumab (J06BB16), motavizumab (J06BB17), raxibacumab (J06BB18), bezlotoxumab (J06BB21), obiltoxaximab (J06BB22), anthrax immunoglobulin (J06BB19), combination (J06BB30), or any mixture thereof. The pharmaceutical formulation according to claim 4. **Claim 6** The pharmaceutical preparation according to claim 4, wherein the biologically active drug is a monoclonal antibody and is selected from edrecolomab (L01XC01), rituximab (L01XC02), trastuzumab (L01XC03), gemtuzumab ozogamicin (L01XC05), cetuximab (L01XC06), bevacizumab (L01XC07), panitumumab (L01XC08), catumaxomab (L01XC09), ofatumumab (L01XC10), ipilimumab (L01XC11), brentuximab vedotin (L01XC12), pertuzumab (L01XC13), trastuzumab emtansine (L01XC14), obinutuzumab (L01XC15), dinutuximab beta (L01XC16), nivolumab (L01XC17), pembrolizumab (L01XC18), blinatumomab (L01XC19), ramucirumab (L01XC21), necitumumab (L01XC22), elotuzumab (L01XC23), daratumumab (L01XC24), mogamulizumab (L01XC25), inotuzumab ozogamicin (L01XC26), orlatumumab (L01XC27), durvalumab (L01XC28), brolucizumab (L01XC29), abemaciclib (L01XC31), atezolizumab (L01XC32), semipramosumab (L01XC33), moxetumomab pasudotox (L01XC34), tafasitamab (L01XC35), enfortumab vedotin (L01XC36), polatuzumab vedotin (L01XC37), isatuximab (L01XC38), belantamab mafodotin (L01XC39), dostarlimab (L01XC40), trastuzumab deruxtecan (L01XC41), alemtuzumab (L04AA34), bispecific T cell engagers (BiTEs, e.g., blinatumomab, solitomab, AMG 330, MT112, MT111, BAY 2010112, MEDI-565), or a mixture of any of these.

7. The biological active drug is an antibody mimetic and is selected from an affibody molecule (such as ABY-025), an affilin (such as SPVF2801), an affimer, an affitin, an alphabody (such as CMPX-1023), an anticalin, an avimer, a designed ankyrin repeat protein (DARPin such as MP0112), a fibronomer, a knotted domain peptide (such as Ecallantide (Kalbitor)), an adnectin, and a monobody (such as Pegaptanib (Macugen)), a nanobody, a single domain antibody such as a camelid antibody, and a V ＮＡＲ fragment, a bivalent single domain antibody (such as caplacizumab (Cablivi)), and an armadillo repeat protein (here, a designed armadillo repeat protein), a peptide aptamer, and a notchin, or a mixture of any of these, the pharmaceutical preparation according to claim 4.

8. The pharmaceutical preparation according to claim 4, wherein the biologically active drug is a human peptide hormone and is selected from amylin, anti-Müllerian hormone, adiponectin, adrenocorticotropic hormone, angiotensinogen, angiotensin, antidiuretic hormone, atrial natriuretic peptide, brain natriuretic peptide, calcitonin, cholecystokinin, corticotropin-releasing hormone, cortistatin, enkephalin, endothelin, erythropoietin, follicle-stimulating hormone, galanin, gastric inhibitory polypeptide, gastrin, ghrelin, glucagon, glucagon-like peptide-1, gonadotropin-releasing hormone, growth hormone-releasing hormone, hepcidin, human chorionic gonadotropin, human placental lactogen, growth hormone, inhibin, insulin, insulin-like growth factor, leptin, lipotropin, luteinizing hormone, melanocyte-stimulating hormone, motilin, orexin, osteocalcin, oxytocin, pancreatic polypeptide, parathyroid hormone, pituitary adenylate cyclase-activating peptide, prolactin, prolactin-releasing hormone, relaxin, renin, secretin, somatostatin, growth hormone-inhibiting hormone, growth hormone release-inhibiting hormone, somatotropin release-inhibiting factor, somatotropin release-inhibiting hormone, thrombopoietin, thyroid-stimulating hormone, thyrotropin, thyrotropin-releasing hormone, vasoactive intestinal peptide, guanilin / uroguanylin, tetracosactide, mecaseremin, somapacitan, pegvisomant, vasopressin, desmopressin, terlipressin, lypressin, ornipressin, argipressin, demoxytocin, carbetocin, gonadorelin, nafarelin, histrelin, octreotide, lanreotide, vapreotide, pasireotide, ganirelix, cetrorelix, elagolix, relugolix, teriparatide, elcatonin.

9. The pharmaceutical preparation according to claim 4, wherein the biologically active drug is a cytokine or a cytokine antagonist and is selected from IL-1 receptor antagonist, anakinra, IL-2, IL-7, IL-15, IL-21, TNF-alpha, interferon-gamma.

10. The pharmaceutical preparation according to claim 1 or 2, further comprising a pharmaceutically acceptable or veterinarily acceptable adjuvant, diluent, or carrier.

11. The pharmaceutical formulation according to claim 1 or 2, wherein the carrier system is an aqueous carrier system.

12. The coating of the inorganic material is (i) zinc oxide (ZnO) and (ii) a mixture with one or more other metal and / or metalloid oxides, The pharmaceutical formulation according to claim 1 or 2, wherein the atomic ratio ((i):(ii)) is between about 1:6 and at most about 6:

1.

13. The formulation according to claim 12, wherein the ratio of zinc oxide to other metal and / or metalloid oxides is from about 1:1 to about 6:

1.

14. The formulation according to claim 12, wherein the ratio of zinc oxide to other metal and / or metalloid oxides is from about 2:1 to about 5:

1.

15. The formulation according to claim 12, wherein the one or more other metal and / or metalloid oxides are selected from aluminum oxide and / or silicon dioxide.

16. The pharmaceutical formulation according to claim 1 or 2, in the form of a sterile injectable dosage form and / or an infusion dosage form.

17. The pharmaceutical formulation according to claim 16, in a form administrable via a surgical administration device for forming a depot formulation.

18. A process for preparing the formulation according to claim 1 or 2, wherein the coated particles are produced by applying a layer of a mixed oxide coating material to the core and / or a previously coated core by atomic layer deposition.

19. An injectable dosage form and / or an infusion dosage form containing the formulation according to claim 1, contained in a reservoir and injection means or infusion means.

20. The dosage form according to claim 19, which is a surgical administration device for forming a depot formulation.

21. The dosage form according to claim 1, wherein the coated particles according to claim 19 and the carrier system are separately housed and mixing occurs before and / or during injection or infusion.

22. Use of the formulation according to claim 1 or the dosage form according to claim 19 for the manufacture of a medicament.

23. The formulation, use or method for use according to claim 22, wherein after injection, the formulation provides a depot formulation in which the API is released over a period of 1 week to about 3 months.

24. The formulation, use or method for use according to claim 23, wherein the API is released over a period of 1 month to 2 months.

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