Vapor deposition techniques for preparing pharmaceutical compositions

Abstract

The present invention provides a process for preparing a pharmaceutical or veterinary composition in the form of a plurality of particles, the process comprising: (a) loading a plurality of solid cores comprising a bioactive agent into a stationary vapor deposition reactor chamber; and (b) applying a vapor deposition technique to enclose, enclose and / or encapsulate the core with one or more layers comprising one or more coating materials, each comprising one or more metal-containing or metalloid-containing compounds; and (c) sequentially repeating the above step (b) as needed to form a plurality of particles having an average diameter based on weight, quantity and / or volume of between about 10 nm and about 100 [mu] m, each particle comprising a respective solid core and a coating surrounding, enclosing and / or encapsulating the core, the vapor deposition technique includes: (1) introducing a pulse of a first reactant gas into the stationary vapor deposition reactor chamber and allowing the first reactant gas to contact the solid core for a predetermined soak time period; (2) after the step (1), evacuating the chamber of the fixed vapor deposition reactor and / or purging the chamber of the fixed vapor deposition reactor with inert gas; (3) introducing a pulse of a second reactant gas into the stationary vapor deposition reactor chamber and allowing the second reactant gas to contact the solid core for a predetermined soak time period; and (4) after step (3), evacuating the stationary vapor deposition reactor chamber and / or purging the stationary vapor deposition reactor chamber with an inert gas wherein the first reactant gas or the second reactant gas comprises a metal-containing or metalloid-containing compound.

Description

Technical Field

[0001] The present invention relates to a novel method for producing a composition useful in the field of drug delivery. Background Art

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

[0003] In the field of drug delivery, the ability to control the drug release profile is of vital importance. It is desirable to ensure that the active ingredient is released in vivo at a desired and predictable rate after administration in order to ensure a more optimized pharmacokinetic profile.

[0004] In the case of sustained release compositions, it is essential that the drug delivery composition provides a release profile showing a minimal initial rapid release of the active ingredient, i.e. high drug concentrations in plasma shortly after administration. In the case of drugs with a narrow therapeutic window or drugs that are toxic at high plasma concentrations, such a 'burst' release can be dangerous.

[0005] In the case of an injectable suspension of the active ingredient, it is also important to control the size of the suspended particles so that they can pass through a needle for injection. If large aggregated particles are present, they will not only clog the needle (through which the suspension is injected), but will also not form a stable suspension within the injection (i.e., they will instead tend to settle to the bottom thereof).

[0006] Therefore, there is a general need in the art for efficient and / or improved drug transport and delivery systems.

[0007] Atomic layer deposition (ALD) is a technique for depositing thin films containing a variety of materials on a solid substrate, including organic materials, biomaterials, polymer materials, and especially inorganic materials, such as metal oxides. It is an enabling technology for atomic and near-atomic scale manufacturing (ACSM) of materials, structures, devices, and systems in a variety of applications (see, for example, Zhang et al. Nanomanuf. Metrol. 2022, https: / / doi.org / 10.1007 / s41871-022-00136-8). Based on its self-limiting characteristics, ALD can achieve atomic-level thickness that is controlled only by adjusting the number of growth cycles. In addition, multiple layers can be deposited, and the characteristics of each layer can be customized at the atomic level.

[0008] Due to its atomic-scale control, ALD is used as a key technology for manufacturing, for example, next-generation semiconductors, or for atomic-scale synthesis of advanced catalysts and precise fabrication of nanostructures, nanoclusters, and single atoms (see, e.g., Zhang et al., supra).

[0009] The technique is typically performed at low pressure and elevated temperature. The film coating is produced by alternating exposure of a solid substrate within the ALD reactor chamber to vaporized reactants in the gas phase. The substrate can be a silicon wafer, granular material, or small particles (e.g., microparticles or nanoparticles).

[0010] The coated substrate is protected by the solid coating from chemical reactions (decomposition) and physical changes.ALD can also potentially be used to control the release rate of substrate materials within a solvent, which makes it potentially useful in the formulation of active pharmaceutical ingredients.

[0011] In ALD, a first precursor (which may be metal-containing) is fed into the ALD reactor chamber (in a so-called 'precursor pulse') and an adsorbed atomic or molecular monolayer is formed at the substrate surface. Excess first precursor is then purged from the reactor, and a second precursor (such as water) is then pulsed into the reactor. This reacts with the first precursor, resulting in the formation of a monolayer, e.g., a metal oxide, on the substrate surface. Subsequent purge pulses are followed by further pulses of the first precursor, and thus a new cycle of the same events (a so-called 'ALD cycle') begins.

[0012] Alternatively, in 'spatial ALD', a separate reactor chamber contains each precursor, and the coated substrate is moved from one reactor chamber to another in order to form the coating. In this or other ALD methods, introducing a precursor to a coated substrate (or vice versa) can be considered equivalent to a 'precursor pulse', and separating a precursor from a substrate to be coated (or vice versa) can be considered equivalent to a 'purge pulse'.

[0013] The thickness of the film coating is controlled, among other things, by the number of ALD cycles performed.

[0014] In a normal ALD process, since only atomic or molecular monolayers are produced in any one cycle, no discernible physical interfaces are formed between these monolayers, which essentially become a continuum at the surface of the substrate.

[0015] In international patent application WO 2014 / 187995 a method is described in which a plurality of ALD cycles are performed and then the resulting coated substrate is periodically removed from the reactor and subjected to a redispersion / agitation step in order to present new surfaces available for precursor adsorption.

[0016] The agitation step is performed primarily to address a problem observed with nanoparticles and microparticles, namely, that during the ALD coating process, particle aggregation occurs, resulting in 'pinholes' formed by contact points between such particles. The redispersion / agitation step is performed by placing the coated substrate in water and subjecting it to sonication, which results in deagglomeration and disruption of contact points between individual particles of the coated active.

[0017] The particles were then loaded back into the reactor, and the ALD coating step of the powder and the step of deagglomerating the powder were repeated 3 times, for a total of 4 series of cycles. This method has been found to allow the formation of coated particles that are largely free of pinholes (see also, Hellrup et al., Int. J. Pharm., 529, 116 (2017)).

[0018] It has been found that the method of performing 'multiple sets' of ALD coating cycles followed by intermittent dispersion, as described in WO 2014 / 187995, produces distinct, separate coating layers defined by distinct, visible physical interfaces between such coating layers. Such interfaces are more distinct than interfaces visible between layers of different coating materials. Interfaces formed by such intermittent dispersions of particles are clearly visible as regions of higher electron permeability by techniques such as transmission electron microscopy (TEM). Similar interfaces are not visible when coatings of the same material are built up from the substrate surface one atomic layer at a time, as explained below.

[0019] As described in international patent application WO 2021 / 111149, we have recently found that it is advantageous to deagglomerate the aggregated particles into primary particles outside the reactor by a dry process involving a combination of a device for applying mechanical force and a sieve, in particular a sonic screening device. This avoids the need to employ aggressive deagglomeration techniques, such as sonication, and the need to dry the particles before returning them to the reactor for further coating. We have found that carrying out the deagglomeration step in this way allows the presentation of coated particles that are substantially completely free of pinholes in a form that can be easily processed into a pharmaceutical formulation.

[0020] As described in unpublished UK patent application number GB 2108305.0, we have recently found that it is advantageous to use a vibratory sieving technique to deagglomerate agglomerated particles. In particular, the vibratory sieving technique results in deagglomerated coated particles being substantially free of cracks through which the active ingredient may be released in an uncontrolled manner.

[0021] In attempting to further scale up the process described in WO 2021 / 111149 and UK Patent Application No. GB ​​2108305.0, we found that when using a larger scale continuous flow reactor, the consistency of the coating between different particles coated together becomes unsatisfactory. This problem has been unexpectedly solved by means of the process described herein. Summary of the invention

[0022] According to a first aspect of the present invention, there is provided a method for preparing a composition in the form of a plurality of particles, the method comprising:

[0023] (a) loading a plurality of solid cores comprising a bioactive agent into a stationary vapor deposition reactor chamber; and

[0024] (b) applying a vapor deposition technique to surround, enclose and / or encapsulate the core with one or more layers comprising one or more coating materials, each of which comprises one or more metal-containing or metalloid-containing compounds; and

[0025] (c) sequentially repeating step (b) as needed to form a plurality of particles having an average diameter based on weight, number and / or volume between about 10 nm and about 100 μm, each particle comprising a corresponding solid core and a coating surrounding, enclosing and / or encapsulating the core,

[0026] The vapor deposition technology includes:

[0027] (1) introducing a pulse of a first reactant gas into a stationary vapor deposition reactor chamber and allowing the first reactant gas to contact the solid core for a predetermined soaking period of time;

[0028] (2) after step (1), evacuating the fixed vapor deposition reactor chamber and / or purging the fixed vapor deposition reactor chamber with an inert gas;

[0029] (3) introducing a pulse of a second reactant gas into the stationary vapor deposition reactor chamber and allowing the second reactant gas to contact the solid core for a predetermined soaking period of time; and

[0030] (4) after step (3), evacuating the fixed vapor deposition reactor chamber and / or purging the fixed vapor deposition reactor chamber with an inert gas,

[0031] The first reactant gas or the second reactant gas includes a metal-containing or metalloid-containing compound.

[0032] In the method of the present invention, both the first reactant gas and the second reactant gas are contacted with the particles for a predetermined soaking period, respectively. The term "soaking" is used in this application only for the sake of clarity, that is, to make the associated predetermined period of time easily distinguishable from other predetermined periods of time that may be mentioned in this application. It should be understood that the term "soaking" in no way limits the associated predetermined period of time.

[0033] A process with such a soaking time increases coating uniformity (also referred to as coating integrity or shell integrity, which can be measured, for example, as described below) because it allows each gas to diffuse conformally in a high aspect ratio substrate (e.g., especially a powder). This is because the substrate has an increased surface area, requiring a longer period of time to waste and react with all available surface sites.

[0034] When the method involves the use of reactants with slow reactivity, the above benefits of including soaking time are even more pronounced, as more time is provided for the reactants to react on the substrate surface. For example, when depositing AlZnO, this is particularly evident for diethylzinc (DEZ), as its probability of reaction toward the surface is lower than, for example, trimethylaluminum (TMA).

[0035] Known ALD methods operate in a continuous flow mode, which means that the pump is actively pumped on the reactor during the entire method, and the gas passes continuously over the powder substrate. In contrast, including a soaking time in the method can prevent the gas from flowing continuously over the powder substrate. For example, introducing a soaking time can include closing a valve to prevent inflow or outflow from the reactor chamber. If the valve is closed to prevent inflow (e.g., the valve is positioned to close the inlet of the reactor chamber), the pressure in the reactor chamber can be substantially constant during the soaking time. However, due to the occurrence of the reaction, the pressure may change somewhat, and the pressure change will depend on the stoichiometry of these reactions. Alternatively, if the valve is closed to prevent outflow (e.g., the valve is positioned to close the outlet of the reactor chamber), the pressure in the reactor chamber can increase steadily during the soaking time as more precursors enter the reactor chamber. This can advantageously promote the precursor to further enter the powder bed and increase the possibility of reacting with the particle surface therein, thereby enhancing the effect of the soaking time.

[0036] ALD processes that include such soak times are sometimes referred to as "stop-flow" processes.

[0037] Inclusion of the soak time not only produces a coating with good shell integrity and a more controlled release profile, but also brings the coating composition closer to the ALD process setup. When using an ALD cycling scheme consisting of three DEZ cycles and one TMA cycle, one would expect to obtain an atomic ratio of 3:1 between Zn and Al in the resulting shell. This is not the case when employing continuous flow deposition, where the atomic ratio is closer to 1:1, since DEZ has a lower probability of reaction than TMA. When using stopped flow, the same ratio is very close to 3:1.

[0038] The method of the present invention also requires the use of a fixed vapor deposition reactor chamber. It should be understood that a fixed reactor chamber in the context of the present invention is a reactor chamber that remains fixed when used to perform vapor deposition techniques, excluding negligible vibrations caused by associated machinery. This is in contrast to a reactor chamber that rotates or vibrates or otherwise actively moves during the vapor deposition process.

[0039] It will be appreciated that the "predetermined time period" of the soaking time may be any suitable time period. The time period depends largely on the particle sample size (or batch size), wherein the smaller the sample size, the shorter the soaking time required, and the larger the sample size, the longer the soaking time required. The soaking time may also depend on factors such as the characteristics of the solid core and / or the type of reactant gas. In addition, the soaking time may depend on the design of the ALD reactor, such as the size of the reaction chamber, the distance to the inlet valve, and the valve to the pump. The soaking time may be in the range of about 2 seconds to about 30 minutes. For example, the soaking time may be about 30 seconds, 1 minute, 3 minutes, 10 minutes, or 15 minutes.

[0040] It will be appreciated by those skilled in the art that for any combination of the above variables, the beneficial effects provided by soaking time are limited. For example, for a given process, the first 5 second soaking period may provide significant benefits to coating uniformity and integrity, the second 5 second soaking period may provide some additional benefits, but less than the benefits provided by the first cycle, and the third 5 second period may be an additional benefit that is almost negligible compared to the benefits already obtained (this is a simplified example for illustrative purposes only). In other words, it is predicted that as soaking time increases, the benefits provided by soaking time can be modeled as a curve that initially increases with increasing high gradients, but as soaking time increases, the gradient will decrease until a steady state is eventually reached, at which point further soaking may no longer be beneficial.

[0041] Although the "benefit" of soaking time is not limited to shell integrity, it should be understood that the shell integrity of the resulting particles can be used as an indicator of the effectiveness of the soaking time. Accordingly, in order to select an appropriate soaking time for a particular process, one skilled in the art can conduct experiments involving gradually increasing soaking times to determine the desired compromise between the length of the soaking time and the benefit it provides for shell integrity. In the simplified example mentioned above, the skilled artisan can determine that a soaking time of approximately 10 seconds is appropriate, as any longer soaking time will observe minimal additional benefits.

[0042] In some examples of the invention, the predetermined soaking time period can be selected to provide a shell integrity, such as at least about 70%, such as at least about 80%, such as at least about 90%, such as at least about 95%, such as about 98.5%, as measured as described below. It should be understood that, as described herein, the shell or coating integrity can be considered to be the reciprocal of the API solubility in a solvent that dissolves the API but does not dissolve the coating material. For example, an API solubility of about 5% indicates a shell / coating integrity of about 95%. Suitable methods for determining coating integrity via determining API solubility are provided in the examples described below.

[0043] During the method of the present invention, the valve at the pump inlet is closed so that the reactant gases reside in the reaction chamber for a predetermined soak time without any active pumping, and then the chamber is pumped. Depending on the reactor, the valve may be completely closed so that no gas passes, or it may not be possible to completely close the valve so that there is a small amount of pumping (e.g., nitrogen carrier gas may still flow in the chamber). In either case, it should be understood that no active pumping occurs. That is, there is substantially no pumping.

[0044] Furthermore, the predetermined soaking period is preferably performed in the substantial absence of mechanical agitation of the plurality of solid cores. Agitation or sieving of the solid cores may be performed during other steps of the method, as further described herein.

[0045] The method can optionally include the multi-pulse of reactant gas, i.e. short inflow burst, without the need to purge / rinse between each multi-pulse, wherein each multi-pulse pumping reactor reaches a predetermined pumping time period. With regard to the amount of the reactant substrate applied to the solid core, this type of multi-pulse is equivalent to a single long pulse of reactant gas. By applying reactant in a multi-pulse mode, a more consistent covering of the solid core is achieved. Each multi-pulse pumping time can be approximately 0.1 to 1000 seconds, approximately 1 to 500 seconds, approximately 2 to 250 seconds, approximately 3 to 100 seconds, approximately 4 to 50 seconds or approximately 5 to 10 seconds, for example 9 seconds. Multi-pulse can apply approximately 5 to 1000 times, approximately 10 to 250 times or approximately 20 to 50 times in a single step.

[0046] It will be well understood by those skilled in the art that the term "solid" includes any form of material that retains its shape and density without restriction, and / or the molecules therein are generally compressed as closely as possible within the limits allowed by the repulsive forces between them. The solid core has at least a solid outer surface on which a layer of coating material can be deposited. The interior of the solid core may also be solid, or may alternatively be hollow. For example, if the particles are spray-dried before they are placed in a reaction vessel, they may be hollow due to the spray-drying technique. In an alternative, the core may comprise an agglomerate of smaller "primary" particles, i.e., secondary particles of a size range defined herein, which are subsequently coated as described herein.

[0047] The method of the present invention is preferably used to prepare a pharmaceutical composition. Accordingly, the composition, and in particular the solid core, comprises a pharmacologically effective amount of a biologically active agent.

[0048] In this aspect, the solid core may consist essentially of or comprise a bioactive agent (hereinafter interchangeably referred to as a 'drug', and an 'active pharmaceutical ingredient (API)' and / or an 'active ingredient'). Bioactive agents also include biopharmaceuticals and / or biologics. Bioactive agents may also include mixtures of different APIs, such as different API particles or particles comprising more than one API.

[0049] By 'essentially comprising' a bioactive agent, we define the solid core as consisting essentially only of the bioactive agent, i.e., it is free of non-biologically active substances, such as excipients, carriers, etc. (see below), and free of other active substances. This means that the core may contain less than about 5%, such as less than about 3%, including less than about 2%, for example less than about 1% of such other excipients and / or active substances.

[0050] Alternatively, a core comprising a biologically active agent may comprise such agent admixed with one or more pharmaceutical ingredients, may comprise a pharmaceutically acceptable excipient, such as an adjuvant, diluent or carrier, and / or may comprise other biologically active ingredients.

[0051] The bioactive agent may be present in a crystalline, partially crystalline and / or amorphous state. The bioactive agent may further include any substance that is solid or convertible to a solid state at about room temperature (e.g., about 18° C.) and about atmospheric pressure, regardless of its physical form. Such agents (and optionally, other pharmaceutical ingredients as mentioned herein) should also remain in solid form when coated, for example, in an ALD reactor, and should not physically or chemically decompose to an appreciable degree (i.e., not more than about 10% w / w) when coated or after being covered by at least one of the coating materials. The bioactive agent may further be presented in combination with another active substance (e.g., mixed or as a complex).

[0052] As used herein, the term 'biologically active agent' or similar and / or related expressions generally refers to any agent or drug that is capable of producing a physiological effect of some sort in a living subject, particularly including mammals, and especially human subjects (patients), whether in a therapeutic or preventive capacity against a particular disease state or condition.

[0053] The biologically active agent can be, for example, selected from analgesics, anesthetics, anti-ADHD agents, appetite suppressants, anti-addiction agents, antibacterial agents, antimicrobial agents, antifungal agents, antiviral agents, antiparasitic agents, antiprotozoal agents, anthelmintics, ectoparasiticides, vaccines, anticancer agents, antimetabolites, alkylating agents, antitumor agents, topoisomerase inhibitors, immunomodulators, immunostimulants, immunosuppressants, anabolic steroids, anticoagulants, antiplatelet agents, anticonvulsants, antidepressants, antidepressants, antidote agents, antihyperlipidemic agents, antigout agents, antimalarials, antimigraine agents, antiparkinsonian agents, antipruritics, antipsoriatic agents, antiemetics, antiobesity agents, antiasthmatic agents, antibiotics, antidiabetics, antiepileptics, anti Fibrinolytics, antihemorrhagic agents, antitussives, antihypertensives, antimuscarinics, antimycobacterials, antioxidants, antipsychotics, antipyretics, antirheumatic agents, antiarrhythmics, antianxiety agents, aphrodisiacs, cardiac glycosides, cardiotonic agents, religious hallucinogens, reassuring drugs, euphoric drugs, appetite stimulants, antithyroid agents, antianxiety sedatives, hypnotic drugs, tranquilizers, astringents, antibacterial agents, beta blockers, calcium channel blockers, ACE inhibitors, angiotensin II receptor antagonists, renin inhibitors, beta-adrenergic receptor blockers, blood products, blood substitutes, bronchodilators, myocardial inotropes, chemotherapeutic agents, coagulants, corticosteroids, cough suppressants, diuretics, delirium drugs, expectorants, Aphrodisiacs, sex hormones, mood stabilizers, mucolytics, neuroprotectants, nootropics, neurotoxins, dopaminergics, antiparkinsonian agents, free radical scavengers, growth factors, fibrates, bile acid sequestrants, cicatrizing agents, glucocorticoids, mineralocorticoids, hemostatics, hallucinogens, hypothalamic-pituitary hormones, immunizing agents, laxatives, antidiarrheals, lipid regulators, muscle relaxants, parasympathomimetics, parathyroid calcitonin, serenic, statins, stimulants, wakefulness agents, decongestants, dietary minerals, bisphosphonates, cough suppressants, ophthalmics, ontologicals, H1 antagonists, H2 antagonists, proton pump inhibitors, prostate adrenalin, radiopharmaceuticals, hormones, sedatives, antiallergic agents, appetite stimulants, steroids, sympathomimetics, thrombolytics, thyroid agents, vasodilators, xanthines, erectile dysfunction improvers, gastrointestinal agents, histamine receptor antagonists, keratolytics, antianginal agents, nonsteroidal anti-inflammatory agents, COX-2 inhibitors, leukotriene inhibitors, macrolides, NSAIDs, nutritional agents, opioid analgesics, opioid antagonists, potassium channel activators, protease inhibitors, anti-osteoporosis agents, cognitive enhancers, anti-urinary incontinence agents, nutritional oils, anti-benign prostatic hypertrophy agents, essential fatty acids, non-essential fatty acids, radiopharmaceuticals, anti-aging therapeutics, vitamins, or any mixture of these drugs.

[0054] The bioactive agent can also be a cytokine, a peptidomimetic, a peptide, a protein, a toxoid, a serum, an antibody, a vaccine, a nucleoside, a nucleotide, a portion of genetic material, a nucleic acid, or a mixture thereof. Non-limiting examples of therapeutic peptides / proteins are as follows: lepirudin, cetuximab, danase alfa, denileukin, etanercept, bivalirudin, leuprolide, alteplase, interferon alpha-n1, darbepoetin alpha, reteplase, epoetin alpha, salmon calcitonin, interferon alpha-n3, pegfilgrastim, sargramostim, secretin, peginterferon alpha-2b, asparaginase, thyrotropin alpha, antihemophilic factor, anakinra, gramicidin D, intravenous immunoglobulin, anistreplase, insulin (conventional), tenecteplase, urokinase, interferon gamma-1b, interferon alpha-2a (recombinant), coagulation factor VIIa, oprellekin, palifermin, Glucagon (recombinant), aldesleukin, botulinum toxin type B, omalizumab, luteinizing hormone alfa, insulin lispro, insulin glargine, collagenase, rasburicase, adalimumab, imiglucerase, abciximab, alpha-1-proteinase inhibitor, pegapagasid, interferon beta-1a, bovine peg deaminase, human serum albumin, eptifibatide, iodinated human serum albumin, infliximab, follicle-stimulating hormone beta, antidiuretic hormone, interferon beta-1b, hyaluronidase, rituximab, basiliximab, muromonab, digoxin immune Fab (sheep), ibritumomab tiuxetan, daptomycin, tositumomab, pegvisomant, botulinum toxin type A, pancreatic lipase, streptokinase, alemtuzumab, arabinoside glucosidase, carlostomumab, laronidase, urofollicle-stimulating hormone, efalizumab, serum albumin, chorionic gonadotropin alfa, antithymocyte globulin, filgrastim, coagulation factor IX, becaplermin, agarase beta, interferon alfa-2b, oxytocin, enfuvirtide, palivizumab, daclizumab, bevacizumab, acitumomab, eculizumab, panitumumab, ranibizumab, idoxurase, alglucosidase alfa, exenatide, mecasermin, pramlintide, galactase, abatacept, teicosatide, adrenocorticotropin, insulin aspart, insulin detemir, insulin glulisine, pegaptanib, nesiritide, thymalfasin, defibrotide, natural interferon alfa / multiferon, glutathione Lizide acetate, preotact, teicoplanin, canakinumab, ipilimumab, sulodexetide, tocilizumab, teriparatide, pertuzumab, linacept, denosumab, liraglutide, semaglutide, exenatide, lixisenatide, albiglutide, dulaglutide, tesiparatide, golimumab, belatacept, buserelin, velaglucerase alfa, temorelin, brentuximab, tadalaglutide alfa, belimumab, aflibercept, chrysanthemum erwinia asparaginase, octoplasmin, glutaric acid, teduglutide, raxicumab, certolizumab, low-protamine insulin, epoetin ζ, obinutuzumab, fibrinolytic enzyme also known as plasmin, follitropin alfa, romiplostim, lucinatan,Natalizumab, aliskiren, ragweed pollen extract, secukinumab, growth hormone (recombinant), drocoquinolone alpha, afacept, OspA lipoprotein, urokinase, abarelix, sermorelin, aprotinin, o-gemtuzumab, satumomab-pentetide, antithrombin alpha, antithrombin III (human), afotaminase alfa, atezolizumab, autologous cultured chondrocytes, belacontan, blinatumomab, C1 esterase inhibitor (human), coagulation factor XIII A-subunit (recombinant), conestat afamicus (conestat alfa), daratumumab, desirudin, elosulfatase alfa, eloumab, fibrinogen concentrate (human), filgrastim-sndz, intrinsic factor, hepatitis B immune globulin, calcitonin human, Clostridium tetani toxoid immune globulin human, rabies virus immune globulin human, Rho(D) immune globulin human, hyaluronidase (human, recombinant), idarucizumab Luqimab, immunoglobulin (human), vedolizumab, ustekinumab, toroclotide alfa, tuberculin purified protein derivative, seromotagmin alfa, selutuximab, serbemidase alfa, furanosidase, ramucirumab, prothrombin complex, porcine alfa, pembrolizumab, peginterferon beta-1a, ofatumumab, octosacizumab, nivolumab, necituzumab, metreleptin, methoxypoly Epoetin beta, mepolizumab, ixekizumab, insulin degludec, insulin (porcine), insulin (bovine), thyroglobulin, anthrax immune globulin (human), anti-inhibitor coagulant complex, brodalumab, C1 lipase inhibitor (recombinant), chorionic gonadotropin (human), chorionic gonadotropin (recombinant), coagulation factor X (human), denutusimab, emtansin alfa, factor IX complex (human), hepatitis A vaccine, human varicella-zoster immune globulin, ibritumomab tiuxetan, levofloxacin, begolotidase, sulfatamine, protein S (human), siprucel-T, somatotropin (recombinant), susceptor alpha and thrombomodulin alfa, and antisense RNA, RNA interfering agents, messenger RNA, transfer RNA, ribosomal RNA, including RNA aptamers, in sarcomeric and synthetic forms.

[0055] Non-limiting examples of drugs that can be used according to the present invention are all-trans retinoic acid (tretinoin), alprazolam, allopurinol, amiodarone, amlodipine, asparaginase, astemizole, atenolol, azathioprine, azetine, beclomethasone, bendamustine, bleomycin, budesonide, buprenorphine, butalbital, capecitabine, carbamazepine, carbidopa, carboplatin, cefotaxime, cephalexin, chlorambucil, cholestyramine, ciprofloxacin, cisapride, cisplatin, clarithromycin, chloramphenicol ... Nitrazepam, clozapine, cyclophosphamide, cyclosporine, cytarabine, dacarbazine, dactinomycin, daunorubicin, diazepam, diclofenac sodium, digoxin, dipyridamole, divalproex sodium, dobutamine, docetaxel, doxorubicin, doxazosin, enalapril, epirubicin, erlotinib, estradiol, etodolac, etoposide, everolimus, famotidine, felodipine, fentanyl citrate, fexofenadine, filgrastim, finasteride, fluconazole, flunisolide, fluorouracil, Flurbiprofen, flurellanal, fluvoxamine, furosemide, gemcitabine, glipizide, glyburide, ibuprofen, ifosfamide, imatinib, indomethacin, irinotecan, isosorbide dinitrate, isotretinoin, isradipine, itraconazole, ketoconazole, ketoprofen, lamotrigine, lansoprazole, loperamide, loratadine, lorazepam, lovastatin, progesterone, mefenamic acid, mercaptopurine, mesna, methotrexate, methylprednisolone, midazolam, mitomycin, mitoxantrone, moxifloxacin statin, mometasone, nabumetone, naproxen, nicergoline, nifedipine, norfloxacin, omeprazole, oxaliplatin, paclitaxel, phenytoin, piroxicam, procarbazine, quinapril, ramipril, edion, rituximab, sertraline, simvastatin, sulindac, sunitinib, temsirolimus, terbinafine, terfenadine, thioguanine, trastuzumab, triamcinolone, valproic acid, vinblastine, vincristine, vinorelbine, zolpidem, or a pharmaceutically acceptable salt of any one of these.

[0056] The composition prepared by the method of the present invention may contain a benzodiazepine such as alprazolam, chlordiazepoxide, clobazam, chlordiazepoxide, diazepam, estazolam, flurazepam, lorazepam, oxazepam, quazepam, temazepam, triazolam, and pharmaceutically acceptable salts of any of these drugs.

[0057] The anesthetics that can also be used in the composition prepared by the method of the present invention can be local or systemic. Local anesthetics that may be mentioned include amilocaine, amitocaine, articaine, benzocaine, benzonatate, bupivacaine, butacaine, butancaine, chloroprocaine, cinchocaine, cocaine, cyclomethicaine, dibucaine, diperidone, dimethocaine, eucaine, etidocaine, hexylcaine, formocaine, fotocaine, hydroxyprocaine, isobucaine, levobupivacaine, lidocaine, mepivacaine, , meprecaine, mebucaine, nicocaine, oxorcaine, oxethazaine, oxybuprocaine, ethoxycaine, phenacaine, piperocaine, piperidicaine, pramoxine, prilocaine, pramoxine, procaine, procainamide, proparacaine, propoxycaine, pyrocaine, quinicaine, ropivacaine, mesocaine, tolicaine, topicaine, or a pharmaceutically acceptable salt of any of these.

[0058] Psychotropic drugs can also be used in the composition prepared by the method of the present invention. Mentionable psychotropic drugs include 5-HTP, acamprosate, agomelatine, alimazine, amphetamine, dextroamphetamine, amisulpride, amitriptyline, amobarbital, amobarbital / seccobarbital, amoxapine, amphetamine, aripiprazole, asenapine, atomoxetine, baclofen, phenperazine, bromoperidone, bupropion, buspirone, butalbital, carbamazepine, chloral hydrate, chlorpromazine, chlorprothixene, citalopram, clomethiazole, clomipramine, clonidine, clozapine, cyclohexyl bar bital / diazepam, cyproheptadine, cytisine, desipramine, desvenlafaxine, dextroamphetamine, dexmethylphenidate, diphenhydramine, sulfiram, divalproex sodium, doxepin, doxylamine, duloxetine, enanthate, escitalopram, eszopiclone, fluoxetine, flupentixol, fluphenazine, fluspirillin, fluvoxamine, gabapentin, phenacetin, guanfacine, haloperidol, hydroxyzine, iloperidone, imipramine, lamotrigine, levetiracetam, levomepromazine, levomilnacipran, dextroamphetamine, lithium salt, lurasidone, melatonin, melperone, meprobamate, methamphetamine, nesalone, methylphenidate, mianserin, mirtazapine, moclobemide, nalmefene, naltrexone, niprazine, nortriptyline, olanzapine, ondansetron, oxcarbazepine, paliperidone, paroxetine, penfluridol, pentobarbital, perazine, percynanchine, perphenazine, phenelzine, phenobarbital, pimozide, pregabalin, promethazine, prothiopentad, protriptyline, quetiapine, rameltriptyline, reboxetine, reserpine, risperidone, chloramphenicol Rubidium, secobarbital, selegiline, sertindole, sertraline, sodium oxybate, sodium valproate, sodium valproate, sulpiride, thioridazine, thiothixene, tianeptine, tizanidine, topiramate, tranylcypromine, trazodone, trifluoperazine, trimipramine, tryptophan, valerian, 2.3:1 ratio of valproic acid, varenicline, venlafaxine, vilazodone, vortioxetine, zaleplon, ziprasidone, zolpidem, zopiclone, zotepine, zuclopenthixol, and pharmaceutically acceptable salts of any of these.

[0059] Antiparkinson's drugs that may be mentioned include levodopa and apomorphine and pharmaceutically acceptable salts thereof.

[0060] Opioid analgesics that can be used in the compositions prepared by the methods of the present invention include buprenorphine, butorphanol, codeine, fentanyl, hydrocodone, hydromorphone, pethidine, methadone, morphine, nomethadone, opium, oxycodone, oxymorphone, pentazocine, tapentadol, tramadol, and pharmaceutically acceptable salts of any of these drugs.

[0061] Opioid antagonists that can be used in the composition prepared by the method of the present invention include naloxone, nalorfen, niconalorphine, diprenorphine, levallorphan, samidorfen, nalodeine, alvimopan, methylnaltrexone, naloxetol, 6β-naltrexol, oprezomib, befoplan, methylsamidorfen, naldemidine (preferably nalmefene, and especially naltrexone) and pharmaceutically acceptable salts of any of these drugs.

[0062] Anticancer agents that may be included in the composition prepared by the method of the present invention include the following: actinomycin, afatinib, all-trans retinoic acid, amsacrine, anagrelide, arsenic trioxide, axitinib, azacitidine, azathioprine, bendamustine, bexarotene, bleomycin, bortezomib, bosutinib, busulfan, cabazitaxel, capecitabine, carboplatin, chlorambucil, cladribine, clofarabine, cytarabine, dabrafenib, dacarbazine, actinomycin, dasatinib, daunorubicin, decitabine, docetaxel, doxifluridine, doxorubicin, epirubicin, epothilone, erlotinib, estramustine, etoposide, everolimus, fludarabine, fluorouracil, gefitinib, guadecitabine, gemcitabine, hydroxyurea, hydroxyurea, idarubicin, Idelalis, ifosfamide, imatinib, irinotecan, ixazomib, cabozantinib, carfilzomib, crizotinib, lapatinib, lomustine, nitrogen mustard, melphalan, mercaptopurine, mesna, methotrexate, mitotane, mitoxantrone, nelarabine, nilotinib, niraparib, olaparib, oxaliplatin, paclitaxel, panobinostat, pazopanib, pemetrexed, pixantrone, ponatinib, procarbazine Hydrazine, regorafenib, ruxolitinib, sonidegi, sorafenib, sunitinib, tegafur, temozolomide, teniposide, thioguanine, tiotepa, topotecan, trabectedin, valrubicin, vandetanib, vemurafenib, venetoclax, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, vismodegib, and pharmaceutically acceptable salts of any of these drugs.

[0063] Such compounds may be used in any of the following cancers: adenoid cystic carcinoma, adrenal cancer, amyloidosis, anal cancer, ataxia-telangiectasia, atypical nevus syndrome, basal cell carcinoma, bile duct cancer, Birt-Hogg Dubé, duct syndrome, bladder cancer, bone cancer, brain tumors, breast cancer (including male breast cancer), carcinoid tumors, cervical cancer, colorectal cancer, ductal carcinoma, endometrial cancer, esophageal cancer, gastric cancer (gastric cancer), cancer), gastrointestinal stromal tumors, HER2-positive breast cancer, islet cell tumors, juvenile polyposis syndrome, kidney cancer, laryngeal cancer, acute lymphocytic leukemia, all types of acute lymphocytic leukemia, acute myeloid leukemia, adult leukemia, childhood leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, liver cancer, lobular carcinoma, lung cancer, small cell lung cancer, Hodgkin's lymphoma, non-Hodgkin's lymphoma, malignant glioma, melanoma, meningioma, multiple myeloma, myelodysplastic syndrome, nasopharyngeal cancer, neuroendocrine tumors, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, pancreatic neuroendocrine tumors, parathyroid cancer, penile cancer, peritoneal cancer, Peutz-Jeghers syndrome, pituitary tumors, polycythemia vera, prostate cancer, renal cell carcinoma, retinoblastoma, salivary gland cancer, sarcoma, Kaposi's sarcoma, skin cancer, small intestine cancer, stomach cancer cancer), testicular cancer, thymoma, thyroid cancer, uterine (endometrial) cancer, vaginal cancer, Wilms tumor.

[0064] Cancers that may be mentioned include myelodysplastic syndrome and subtypes such as acute myeloid leukemia, refractory anemia or refractory anemia with ringed sideroblasts (if accompanied by neutropenia or thrombocytopenia or requiring transfusions), refractory anemia with excess blasts, refractory anemia with excess blasts in transformation, and chronic myeloid (myelomonocytic) leukemia.

[0065] Osteoporosis drugs that may be mentioned include bisphosphonates such as clodronate, ibandronate, pamidronate, zoledronate, etidronate, alendronate, risedronate, tiludronate, bandronate and derivatives (eg acid derivatives of these compounds).

[0066] Other drugs that may be mentioned for use in the composition prepared by the method of the present invention include immunomodulatory imide drugs (such as thalidomide and its analogs, such as pomalidomide, lenalidomide and apremilast) and pharmaceutically acceptable salts of any of these drugs. Other drugs that may be mentioned include angiotensin II receptor type 2 agonists, such as compound 21 (C21; 3-[4-(1H-imidazol-1-ylmethyl)phenyl]-5-(2-methylpropyl)thiophene-2-[(N-butoxycarbamate)-sulfonamide] and pharmaceutically acceptable (e.g. sodium) salts thereof.

[0067] Preferred anticancer agents include lenalidomide, which is useful for treating multiple myeloma and anemia in low to intermediate risk myelodysplastic syndrome, and especially azacitidine, which is useful for treating certain subtypes of myelodysplastic syndrome. Another specific anticancer drug that may be mentioned is cisplatin, which is a chemotherapeutic agent that can be used for a variety of cancers, including testicular, cervical, ovarian, bladder, lung, esophageal and head and neck cancers, as well as brain tumors, neuroblastoma and mesothelioma.

[0068] Other preferred bioactive agents that may be mentioned include liraglutide, which can be used to treat type 2 diabetes and prevent cardiovascular complications associated with diabetes. Specific drugs that may be mentioned in this regard include glucagon-like peptide-1 receptor agonists, such as exenatide, lixisenatide, albiglutide, dulaglutide, more preferably tezetide and semaglutide, and especially liraglutide.

[0069] Alternatively, instead of (or in addition to) a bioactive agent, the composition prepared by the method of the invention may also contain a diagnostic agent (i.e., an agent that does not itself have direct therapeutic activity but can be used for the diagnosis of a disorder, such as a contrast agent for biological imaging).

[0070] The compositions prepared by the methods of the invention may induce an inflammatory response upon injection, for example, subcutaneous injection. Such a response may be produced by any component or combination of components of such formulations, including coatings or carrier systems.

[0071] Bioactive agents that may be particularly mentioned in this regard include those wherein the bioactive agent, either by itself or in the form of a composition prepared by the method of the invention, when administered to a patient can produce an inflammatory response, or can be expected to produce such a response.

[0072] In this regard, biologically active agents that may be particularly mentioned for use in the compositions prepared by the present invention include, for example, anti-tumor agents, topoisomerase inhibitors, immunomodulators (such as thalidomide, pomalidomide, lenalidomide and apremilast), immunostimulants, immunosuppressants, chemotherapeutic agents, growth factors, vasodilators and radiopharmaceuticals.

[0073] Specific biologically active agents that may be mentioned in this regard include any one or more of the specific anticancer agents listed above, and in particular actinomycin, azacitidine, azathioprine, bendamustine, bexarotene, bleomycin, bortezomib, bosutinib, busulfan, cabazitaxel, capecitabine, carboplatin, chlorambucil, cladribine, clofarabine, cytarabine, dabrafenib, dacarbazine, dactinomycin, daunorubicin, decitabine, docetaxel, doxifluridine, doxorubicin, epirubicin, epothilones, estramustine, etoposide, everolimus, fludarabine, fluorouracil , guadecitabine, gemcitabine, hydroxyurea, idarubicin, ifosfamide, irinotecan, ixazomib, carfilzomib, lomustine, nitrogen mustard, melphalan, mercaptopurine, mesna, methotrexate, mitotane, mitoxantrone, nelarabine, oxaliplatin, paclitaxel, panobinostat, pemetrexed, pixantrone, procarbazine, fluridine, temozolomide, teniposide, thioguanine, thiotepa, topotecan, trabectedin, valrubicin, venetoclax, vinblastine, vincristine, vindesine, vinflunine and vinorelbine, and pharmaceutically acceptable salts of any of these.

[0074] Other bioactive agents that may be mentioned in this regard include certain cytokines, proteins and vaccines, as well as therapeutic peptides / proteins such as daratumumab, isatuximab and complement C1 esterase inhibitors

[0075] Other drugs that may be mentioned in this regard include bendamustine, bleomycin, carboplatin, chlorambucil, cisplatin, cyclophosphamide, cyclosporin, cytarabine, dacarbazine, dactinomycin, daunorubicin, docetaxel, doxorubicin, epirubicin, etoposide, everolimus, fluorouracil, gemcitabine, ifosfamide, irinotecan, mercaptopurine, mesna, methotrexate, midazolam, mitomycin, oxaliplatin, paclitaxel, procarbazine, temsirolimus, thioguanine, vinblastine, vincristine, vinorelbine, or a pharmaceutically acceptable salt of any of these.

[0076] This mild inflammatory response can be alleviated by co-administering an anti-inflammatory agent suitable for injection.

[0077] Suitable anti-inflammatory agents that may be used in this regard include butylpyrazolidines (such as phenylbutazone, mofebutazone, oxybenbutazone, clofebutazone, kebuzone and vinbutazone); acetic acid derivatives and related substances (indomethacin, sulindac, tolmetin, zomepirac, diclofenac, alclofenac, bumadinone, etodolac, lonarazoc, fentisa, acemetacin, phenpyramine, oxametacin, proglumetacin, ketorolac, aceclofenac and bufenac); oxicams (such as piroxicam, tenoxicam, droxicam, lornoxicam and meloxicam); propionic acid derivatives (such as ibuprofen, naproxen, ketoprofen, fenoprofen, fenbufen, benoprofen, suprofen, pirprofen, flurbiprofen, indoprofen, tiaprofenic acid, oxaprozin, ibuprofen, dexibuprofen, flunoprofen, amiprofen, dexketoprofen, vidaprofen, carprofen and tepoxalin); fenamates (such as mefenamic acid, tolfenamic acid, flufenamic acid, meclofenamic acid and flunixin), coxibs (such as celecoxib, rofecoxib, valdecoxib, parecoxib, etoricoxib, romiracoxib, firocoxib, robenacoxib, mavalacoxib and cimicoxib); other nonsteroidal anti-inflammatory drugs (such as nabumetone, niflumic acid, azaprozone, glucosamine, benzylamine, glucosamine polysulfate, bevacizumab, orgotin, nimesulide, fepiridone, diacerein, moniflumate, tenidab, oxaprodol, chondroitin sulfate, pentosan polysulfate and aminopropionitrile);Corticosteroids (such as 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 (Purbedilon), aldosterone, corticosterone (17-deoxycortisol), cortisol (hydrocortisone), cortisone, pregnenolone, progesterone, fluprogesterone (fluprogesterone), fluorometholone, Medroxyprogesterone (hydroxymethylprogesterone), prednisolone acetate (21-acetoxypregnenolone), cloprednisone, cloprednol, difluprednate, fludrocortisone, fluocinolone, fluperonolone, fluprednisolone, loteprednol, methylprednisolone, prednicarbate, prednisolone, prednisone, tesocotor, triamcinolone, alclometasone, beclomethasone, betamethasone, clobetasol, clobetasone, clocortolone, desomethasone, dexamethasone, diflorasone, diflucortol, fluclorol, flumethasone, flucortolone, fluprednidine, fluticasone, fluticasone furoate, halometasone, methylprednisolone, mometasone, mometasone furoate, paramethasone, prednisone, rimexolone, urobetasol (halobetasol), amcinonide, budesonide quinoline (such as hydroxycinchophenone); gold preparations (such as sodium aurothiomalate, sodium aurothiosulfate, auranofin, aurothioglucose, aurothiopropanol); penicillamine and similar drugs (such as bucillamine); and antihistamines (such as akvastine, alimamazine, antazoline, astemizole, azastatin, azelastine, bamipine, bilastine, bromophenadrine, bromophenamine, buclizine, cetirizine, cinnarizine, cyclizine, cyproheptadine, detropine, desloratadine, dexbromophenamine, dextrochlorpheniramine amine, diphenylpyrroline, dimenhydrinate, dimetane, doxylamine, ebastine, epinastine, phenidamine, pheniramine, fexofenadine, histidine, hydroxyprometazine, isothiol, carbinoxamine, ketotifen, gifenadine, clemastine, clociclizine, chlorphenamine, chlorphenoxamine, clopyramine, levocetirizine, loratadine, mebuhydrine, meketazine, meclozine, mepyramine, mepyrine, medipramine, mizolastine, oxatomide, oxomezine, pimetidine, promezine, pyrobutamine, rupadine, secfenadine, talastine, tenadine, terfenadine, thiazinamide, thiazine, toluenesulfonamide, triphenamide, tripnamine, triprolidine and triptovaline). Any one or more of the above anti-inflammatory agents may be used in combination. ;

[0078] Preferred anti-inflammatory agents include nonsteroidal anti-inflammatory drugs such as diclofenac, ketoprofen, meloxicam, aceclofenac, flurbiprofen, parecoxib, ketorolac, indomethacin or pharmaceutically acceptable salts thereof.

[0079] The subject may receive (or may have received) one or more of the above-described co-therapeutic agents and / or anti-inflammatory agents separately from the composition prepared by the method of the invention, meaning that one or more of those other therapeutic agents are received at a prescribed dose before, in addition to, and / or after treatment with the composition prepared by the method of the invention.

[0080] 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 administered separately (simultaneously or sequentially) in different formulations (hereinafter referred to as a "combination product").

[0081] Such combination products provide for the combined administration of a biologically active agent and an anti-inflammatory agent, and thus can be presented as separate formulations, wherein at least one of those formulations is a composition prepared by the methods of the invention and at least one comprises the anti-inflammatory agent in the separate formulation; or can be presented (i.e., formulated) as a combined preparation (i.e., presented in a single formulation that includes the biologically active agent and the anti-inflammatory agent).

[0082] In this regard, the anti-inflammatory agent may be co-present with the bioactive agent in appropriate doses in one or more cores forming part of the composition prepared by the methods of the invention as described above, or may be formulated using the same or similar methods for coating as described above for the bioactive agent, including the methods of the invention, which may allow release of the other anti-inflammatory agent on the same or different time scales.

[0083] Pharmaceutically acceptable salts of bioactive agents include acid addition salts and base addition salts. Such salts can be formed by conventional means, for example, by reacting the free acid or free base form of the compound of the present invention with one or more equivalents of a suitable acid or base, optionally in a solvent or in a medium in which the salt is insoluble, and then removing the solvent or the medium using standard techniques (e.g., in a vacuum, by freeze drying or by filtering). Salts can also be prepared using techniques known to those skilled in the art, such as by exchanging the counter ions of the compound of the present invention in salt form with another counter ion (e.g., using a suitable ion exchange resin).

[0084] Specific salts that may be mentioned include acid addition salts of, for example, hydrochloric acid, L-lactic acid, acetic acid, phosphoric acid, (+)-L-tartaric acid, citric acid, propionic acid, butyric acid, caproic acid, L-aspartic acid, L-glutamic acid, succinic acid, ethylenediaminetetraacetic acid (EDTA), maleic acid, methanesulfonic acid, and the like.

[0085] The composition prepared by the method of the present invention may contain a pharmacologically effective amount of a biologically active agent. The term 'pharmacologically effective amount' refers to the amount of such active ingredient: whether administered alone or in combination with another active ingredient, the active ingredient is capable of imparting a desired physiological change (such as a therapeutic effect) to the patient being treated. Such biological or medical responses or such effects in the patient can be objective (i.e., measurable by a test or marker) or subjective (i.e., the subject gives an indication or feeling of the effect), and include at least partial relief of the symptoms of the disease or disorder being treated or curing or preventing the disease or disorder.

[0086] Therefore, the dose of active ingredient that can be administered to a patient should be sufficient to affect a therapeutic response within a reasonable and / or relevant time frame. Those skilled in the art will recognize that the selection of the exact dose and composition and the most appropriate delivery regimen will also be affected not only by the nature of the active ingredient, but also, among other things, by the pharmacological properties of the formulation, the route of administration, the nature and severity of the condition being treated, and the physical condition and mental acuity of the recipient, as well as the age, condition, weight, sex and response of the patient being treated, and the stage / severity of the disease, as well as genetic differences between patients.

[0087] Administration of the composition prepared by the method of the present invention may be continuous or intermittent (eg by bolus injection). The dosage of the active ingredient may also be determined by the time and frequency of administration.

[0088] In any event, a medical practitioner or other skilled artisan will be able to routinely determine the actual dosage of any particular active ingredient that will be most suitable for an individual patient.

[0089] According to the present invention, the non-biologically active adjuvants, diluents and carriers that can be used in the core to be coated can include pharmaceutically acceptable substances soluble in water, such as carbohydrates, for example sugars, such as lactose and / or trehalose, and sugar alcohols, such as mannitol, sorbitol and xylitol; or pharmaceutically acceptable inorganic salts, such as sodium chloride. Preferred carrier / excipient materials include sugars and sugar alcohols. When the bioactive agent is a complex macromolecule, such as a peptide, protein or part of genetic material, etc. (for example, as generally described, and / or the specific peptide / protein described above (including vaccines)), such carrier / excipient materials are particularly useful. Embedding complex macromolecules in excipients in this way usually produces a larger core for coating, and thereby produces larger coated particles.

[0090] In addition to comprising one or more biologically active agents, the core may comprise one or more non-biologically active adjuvants, diluents and carriers including emollients and / or other excipients with functional properties, such as buffers and / or pH adjusters (eg, citric acid).

[0091] When injected, the preparation produced by the method of the present invention provides a reservoir preparation from which the bioactive agent is released over an extended period of time. The period of time can be at least about 3 days, such as about 5 days or about 7 days, and is up to a period of about one year, such as 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).

[0092] The solid core is provided in the form of nanoparticles or more preferably microparticles. The preferred average diameter based on weight, number or volume is between about 50nm (e.g., about 100nm, such as about 250nm) and about 30μm, for example, between about 500nm and about 100μm, more particularly between about 1μm and about 50μm, such as about 25μm, for example, about 20μm.

[0093] As used herein, the term 'mean diameter based on weight' will be understood by the skilled person to include that the average particle size is characterized and defined by a particle size distribution by weight, i.e., a distribution in which the existing fraction (relative amount) in each size class is defined as the weight fraction, as obtained by, for example, sieving (e.g., wet sieving). As used herein, the term 'mean diameter based on number' will be understood by the skilled person to include that the average particle size is characterized and defined by a particle size distribution by number, i.e., a distribution in which the existing fraction (relative amount) in each size class is defined as the number fraction, as measured by, for example, microscopy. As used herein, the term 'mean diameter based on volume' will be understood by the skilled person to include that the average particle size is characterized and defined by a particle size distribution by volume, i.e., a distribution in which the existing fraction (relative amount) in each size class is defined as the volume fraction, as measured by, for example, laser diffraction. The skilled person will also understand that there are other suitable ways of expressing the average diameter, such as the average diameter based on area, and these other expressions of the average diameter are interchangeable with those used herein. Other instruments known in the art to measure particle size may be employed, such as equipment sold by, for example, Malvern Instruments, Ltd (Worcestershire, UK) and Shimadzu (Kyoto, Japan).

[0094] The solid core may have a preferred average diameter based on weight, number or volume between about 50 nm (e.g., about 100 nm, such as about 250 nm) and about 30 μm, for example between about 500 nm and about 100 μm, more particularly between about 1 μm and about 50 μm, such as about 25 μm, for example about 20 μm.

[0095] In the method of the present invention, at least 200 mg, optionally at least 1 g or at least 10 g of solid cores may be loaded into a stationary vapor deposition reactor chamber for the vapor deposition technique to be applied.

[0096] The particles can 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, and especially less than about 2, and / or can have a radius variation of at least about 90% of the particles (measured from the center of gravity to the surface of the particle) that does not exceed about 50% of the average, such as does not exceed about 30% of this value, for example does not exceed about 20% of this value.

[0097] However, it is also possible to coat particles in any shape according to the invention. For example, particles of irregular shape (e.g. 'raisin' shape), needle-shaped, flake-shaped or cuboid-shaped may be coated. For non-spherical particles, the size may be indicated, for example, as the size of a corresponding spherical particle of the same weight, volume or surface area. Hollow particles and particles with holes, cracks etc., such as fibrous particles or 'tangled' particles, may also be coated according to the invention.

[0098] The particles can be obtained in a form suitable for coating, or in such a form, for example by a particle size reduction process (e.g., crushing, cutting, milling or grinding) to a specified weight-based average diameter (as defined above), for example by wet milling, dry milling, air jet milling (including cryogenic micronization), ball milling, such as planetary ball milling, and using end roller mills, roller mills, vibrating mills, hammer mills, roller mills, fluid energy mills, pin mills, etc. Alternatively, the particles can be prepared directly into a suitable size and shape, for example by spray drying, freeze drying, spray freeze drying, vacuum drying, precipitation, including the use of supercritical fluids or other top-down methods (i.e., reducing the size of large particles by, for example, grinding) or bottom-up methods (i.e., increasing the size of particles by, for example, sol-gel techniques, crystallization, etc.). Alternatively, nanoparticles can be prepared by well-known techniques (such as gas condensation, attrition, chemical precipitation, ion implantation, pyrolysis, hydrothermal synthesis, etc.).

[0099] It may be necessary (depending on the manner in which the particles comprising the core are initially provided) to wash and / or clean them to remove impurities that may be derived from their production and then to dry them. Drying can be performed by means of a variety of techniques known to those skilled in the art, including evaporation, spray drying, vacuum drying, freeze drying, fluidized bed drying, microwave drying, IR radiation, drum drying, etc. If dried, the core can be depolymerized by grinding, screening, grinding and / or dry ultrasonic treatment. Alternatively, the core can be treated to remove any volatile substances that may be absorbed onto its surface, for example, by exposing the particles to a vacuum and / or elevated temperature.

[0100] 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 application of a plasma treatment, so as to generate oxygen free radicals at the core surface. This in turn may generate favorable adsorption / nucleation sites on the core for reactants (hereinafter interchangeably referred to as "precursors") for depositing the coating material by vapor deposition techniques.

[0101] More than one layer of coating material is sequentially applied to the core. Preferred vapor deposition techniques include ALD or related techniques such as atomic layer epitaxy (ALE), molecular layer deposition (MLD; a technique similar to ALD, except that molecules (usually organic molecules) are deposited in each pulse rather than atoms), molecular layer epitaxy (MLE), chemical vapor deposition (CVD), atomic layer CVD, molecular layer CVD, physical vapor deposition (PVD), sputtering PVD, reactive sputtering PVD, evaporative PVD, and binary reaction sequence chemistry. ALD is a preferred coating method according to the present invention.

[0102] When ALD is employed, the coating material can be prepared by feeding a precursor or reactant into the ALD reactor chamber (in a so-called 'precursor pulse') to form an adsorbed atomic or molecular monolayer at the particle surface. A subsequent purge pulse is followed by a second precursor pulse into the reactor, which reacts with the first precursor, resulting in the formation of a monolayer of the compound on the substrate surface. Another purge pulse is followed by a further pulse of the first precursor, and thus a new cycle of the same events begins, which is an ALD cycle.

[0103] In most cases, the first reaction in the series of reactions will involve some functional groups or free electron pairs or free radicals of the surface to be coated, such as hydroxyl (-OH) or primary or secondary amino groups (-NH or -NHR, where R is, for example, an aliphatic group, such as an alkyl group). Each reaction is advantageously performed separately and under conditions such that all excess reagents and reaction products are substantially removed before carrying out the subsequent reaction.

[0104] Two or more separate layers or coating materials (also referred to herein as 'coatings' or 'shells', all of which terms are used interchangeably herein) are applied (i.e. 'applied separately') to a solid core comprising a biologically active agent. Such 'separate application' of 'separate layers, coatings or shells' means that the solid core is coated with a first layer of coating material, which layer is formed by more than one (e.g., a plurality or a group) of cycles as described herein, each cycle producing a single layer of coating material, and the resulting coated core may then be subjected to some form of sieving step, such as a vibratory sieving technique, step or method as described herein.

[0105] In other words, a 'vapor deposition (e.g. ALD) cycle' may be repeated several times to provide a 'vapor deposition (e.g. ALD) group' of cycles, which may consist of, for example, 10, 25 or 100 cycles. However, after the group of cycles, the coated cores may be subjected to some form of sieving step, such as a vibratory sieving technique, step or method as described herein, followed by another group of cycles.

[0106] The method can be repeated multiple times as needed, and in this respect, the number of discrete layers of coating materials as defined herein corresponds to the number of these intermittent screening steps. Optionally, at least one of these screening steps includes a vibration screening step. In some instances, at least the final screening step includes a vibration screening step performed before applying the final layer (circulation group) of coating materials. In further instances, more than one (including each) screening step in the screening step includes a vibration screening technique, step or method as described herein.

[0107] The vibrating sieving technique may include a vibrating motor coupled to a sieve and providing a means of vibrating to force the solid product material formed by coating the core through the sieve, which may be located inside or (preferably) outside (i.e., outside) the reactor and configured to deagglomerate any particle aggregates under the vibration force of the coated core before applying a second and / or further layer of coating material. The method may be repeated as many times as desired and / or appropriate before applying a final layer of coating material.

[0108] The vibration forcing tool may include a vibration motor coupled to the screen. The vibration motor is configured to vibrate and / or rotate when power is supplied to it. For example, the vibration motor may be a piezoelectric vibration motor containing a piezoelectric material that changes shape due to the inverse piezoelectric effect when an electric field is applied. The change in shape of the piezoelectric material causes sonic or ultrasonic vibrations of the piezoelectric vibration motor.

[0109] Alternatively, the vibration motor may be an eccentric rotating mass (ERM) vibration motor, which includes a mass that rotates when power is supplied to the motor. The mass deviates from the axis of rotation, causing the motor to be unbalanced and vibrate and / or gyrate due to the rotation of the mass. In addition, the ERM vibration motor may include a plurality of masses positioned at different locations relative to the motor. For example, the ERM vibration motor may include a top mass and a bottom mass, each mass positioned at opposite ends of the motor. By changing each mass and its angle relative to another mass, the vibration and / or gyration of the ERM vibration motor may be changed.

[0110] The vibration motor can be coupled to the screen in a manner wherein the vibration and / or rotation of the motor is transferred to the screen when power is supplied thereto.

[0111] The sieve and the vibrating motor can be suspended from a mounting member (e.g., such as a frame positionable on the floor) via a suspension device so that the sieve and the motor are free to vibrate relative to the mounting member, and the vibration is not substantially transmitted to the mounting member or damped by the mounting member. This allows the vibrating motor and the sieve to vibrate and / or rotate without obstacles, and also reduces the noise generated in the vibratory screening method. The suspension device can include one or more springs or bellows (i.e., air cushions or equivalent buffering devices) that couple the sieve and / or the motor to the mounting member. Manufacturers of vibrating screens or screening machines suitable for carrying out this process include, for example, Russell Finex, SWECO, Filtra Vibracion, VibraScreener, Gough Engineering, and FarleyGreene.

[0112] Preferably, the vibratory screening technique further comprises controlling a vibratory probe coupled to the sieve. The vibratory probe may be controlled to vibrate the sieve at a frequency different from the frequency of vibration caused by the vibratory motor. Preferably, the vibratory probe causes the sieve to vibrate at a higher frequency than the vibration caused by the vibratory motor, and more preferably, the frequency is in the ultrasonic range.

[0113] Providing additional vibration to the screen by means of a vibrating probe reduces the occurrence of blockages in the screen, reduces the likelihood of overloading the screen and reduces the amount of time required to clean the mesh of the screen.

[0114] When each sieving step does not include such a vibratory sieving technique, step, the sieving step can still be performed by one or more other means of forcing the coated mass to pass through the sieve manually, mechanically and / or automatically. The mechanical force can take the form of tapping, oscillation, application of a pressure gradient (e.g., a jet), horizontal rotation, mechanized periodic displacement of the sieve, centrifugal force, sieving, or a combination thereof (such as oscillation and tapping, rotation and tapping, etc.).

[0115] Such alternative force-applying means are preferably mechanical, and may also be vibratory, wherein a suitable alternative means of applying a vibratory force (i.e., a means not including a vibrating motor coupled to a sieve) forces the coated powder mass to pass through the mesh or sieve. Alternative mechanical means of generating oscillations around a balance point may include sound waves (including sonic and ultrasonic waves), or may be mechanical (e.g., tapping) or other means, including combinations thereof (such as ultrasonic and sonic, sonic and tapping, ultrasonic and tapping, etc.).

[0116] In such cases we prefer that at least one of these alternative mechanical sieving steps is performed with the aid of a sonic sieve, as described hereinafter. Manufacturers of suitable sonic sieves include Advantech Manufacturing, Endecott and Tsutsui.

[0117] Preferably, the vibratory sieving technique comprises sieving the coated granules at a throughput of at least 1 g / min. More preferably, the vibratory sieving technique comprises sieving the coated granules at a throughput of 4 g / min or more.

[0118] The flux depends on the screen area, the mesh size of the screen, the particle size, the viscosity of the particles, the static properties of the particles. By combining some of these features, much higher fluxes can be achieved. Therefore, the vibratory sieving technique may more preferably include sieving the coated particles at a flux of up to 1 kg / minute or even higher.

[0119] Any of the above fluxes represent a significant improvement over the use of known mechanical sieving or screening techniques. For example, we have found that sonic screening involves screening in a 15 minute period with a 15 minute cooling period in between, which is necessary to protect the equipment. In order to screen 20 g of coated particles, 9 sets of 15 minutes of active screening time are required, i.e., a total time (including cooling) of 255 minutes. In contrast, by using the vibratory screening technology necessary for the method of the present invention, 20 g of coated particles can be continuously screened in a maximum of 20 minutes or more preferably only 5 minutes or less.

[0120] The mesh size may be determined such that the ratio of the size of the sieved or screened particles to the mesh size of the sieve is about 1:>1, preferably about 1:2, and optionally about 1:4. The mesh size may range from about 20 μm to about 100 μm, preferably from about 20 μm to about 60 μm.

[0121] Suitable screen meshes may include perforated plate, microporous plate, grid, diamond mesh, fine wire, polymer or wire (woven wire mesh), but are preferably formed from metal such as stainless steel.

[0122] Surprisingly, the use of stainless steel mesh in such vibratory sieving techniques is just as gentle on particle coating as the use of softer polymer sieves as part of a mechanical sieving technique such as sonic screening.

[0123] Furthermore, a known problem with sieving powders is the potential danger of generating static electricity. Steel mesh has the advantage of removing static electricity from the powder, which is not the case with polymer mesh, which must be used in sonic sieving.

[0124] In addition, the mesh size of known sonic sieves is limited to about 100 μm because the sound waves travel through the mesh rather than vibrating it. For vibratory screening techniques, this limitation does not exist because there is no reliance on sound waves to generate vibrations in the sieve. Therefore, the vibratory screening technique as described herein allows larger particles to be screened out than using alternative mechanical screening techniques.

[0125] If the (e.g. vibrating) screen is located outside (i.e. on the outside) of the reactor, step (2) of the method of the present invention comprises discharging the coated particles from the vapor deposition reactor and then agitating the coated particles, and step (3) comprises reintroducing the deagglomerated coated particles from step (2) into the vapor deposition reactor and then applying another layer of at least one coating material to the reintroduced particles.

[0126] Alternatively, the coated cores may also be subjected to the aforementioned vibration screening step internally without being removed from the apparatus by means of a continuous process. Such a process would involve applying a vibration force to the solid product mass (which is formed by coating the cores) to pass through a sieve located within the reactor, and being configured to deagglomerate any particle aggregates after the vibration screening of the coated cores by means of a force-applying device within the reactor before being subjected to a second and / or further coating. The process is continued as many times as necessary and / or appropriate before applying a final coating as described herein.

[0127] Having the sieve inside the reaction vessel means that the coating can be applied by means of a continuous process that does not require the particles to be removed from the reactor. Therefore, no manual handling of the particles is required, and no external machinery is required to deagglomerate the aggregated particles. This not only greatly reduces the time to perform the coating process, but is also more convenient and reduces the risk of handling hazardous (e.g., toxic) materials by personnel. It also enhances the reproducibility of the process by limiting manual labor and reduces the risk of contamination.

[0128] We have found that applying separate layers of coating material after external depolymerization results in visible and discernible interfaces as regions of higher electron permeability, which interfaces can be observed by analyzing the coated particles according to the invention and by, for example, TEM. In this respect, the thickness of the layer between the interfaces directly corresponds to the number of cycles in each series performed within the ALD reactor and between the various external agitation steps.

[0129] Since in the ALD coating process the coating occurs at the atomic level, such clear physical interfaces are usually more difficult to observe.

[0130] Without being limited by theory, it is believed that removing the coated particles from the vacuum conditions of the ALD reactor and exposing the newly coated surface to the atmosphere results in structural rearrangement due to relaxation and remodeling of the outermost atomic layers. Such processes are believed to involve rearrangement of surface (and near-surface) atoms, which is driven by a thermodynamic tendency to reduce surface free energy.

[0131] Additionally, surface adsorption of species (e.g. hydrocarbons which are always present in air) may contribute to this phenomenon, as may surface modifications due to reactions of coatings formed with hydrocarbons with atmospheric oxygen, etc. Therefore, if such interfaces are chemically analyzed, they may contain traces of contaminants or core material, such as API forming part of the core, which did not originate from the coating process, such as ALD.

[0132] Whether performed inside or outside the reactor, the particle aggregates are broken up by a device that applies a vibrating force that forces them to pass through a sieve, thereby separating the aggregates into individual particles or aggregates of a desired and predetermined size (and thereby achieving deagglomeration). With respect to the latter, in some cases, the individual primary particle size is so small (i.e. <1 μm) that it is not possible to achieve 'complete' deagglomeration (i.e., where the aggregates are decomposed into individual particles). Instead, deagglomeration is achieved by breaking down larger aggregates into smaller secondary particle aggregates of a desired size, as indicated by the mesh size of the sieve. The smaller aggregates are then coated by a gas phase technique to form fully coated 'particles' in the form of small aggregate particles. In this way, when referring to particles that have been deagglomerated and coated in the context of the present invention, the term 'particles' refers to both individual (primary) particles and aggregated (secondary) particles of a desired size.

[0133] In any case, the desired particle size is maintained (whether individual particles or aggregates of the desired size), and further, continued application of the gas phase coating mechanism to the particles following such deagglomeration via vibratory sieving means that a complete coating is formed on the particles, thereby forming fully coated particles (either individual particles or aggregates of the desired size).

[0134] Whether performed inside or outside the reactor, the method of the present invention can be performed in a manner involving performing the above-mentioned repeated coating and deagglomeration / agitation method of the method for the following number of times: at least 1 time, preferably 2 times, more preferably 3 times, such as 4 times, including 5 times, more particularly 6 times, for example 7 times, and no more than about 100 times, for example no more than about 50 times, such as no more than about 40 times, including no more than about 30 times, such as between 2 and 20 times, for example between 3 and 15 times, such as 10 times, for example 9 times or 8 times, more preferably 6 times or 7 times, and particularly 4 times or 5 times.

[0135] Whether performed inside or outside the reactor, it is preferred that at least one sieving step is performed, and it is further preferred that the step preferably comprises a vibrating sieving step as described above. It is further preferred that at least the final sieving step comprises a vibrating sieving step performed before applying the final layer (circulation group) of coating material. However, it is further preferred that more than one (including each) of the sieving steps comprises a vibrating sieving technique, step or method as described herein.

[0136] These steps are preferably repeated to further enhance the throughput of any vibratory screening technique.

[0137] The total thickness of the coating (which means all individual layers / coatings / shells) will on average be in the region between about 0.5 nm and about 2 μm.

[0138] The minimum thickness of each individual layer / coating / shell will on average be in the region of about 0.1 nm (eg about 0.5 nm, or about 0.75 nm, such as about 1 nm).

[0139] The maximum thickness of each individual layer / coating / shell will depend on the size of the core (firstly), and thereafter on the size of the core with the coating previously applied, and may on average be about 1 percent of the average diameter (i.e. average diameter based on weight, number or volume) of the core or cores with the coating previously applied.

[0140] Preferably, for particles having an average diameter between about 100 nm and about 1 μm, the total coating thickness should average between about 1 nm and about 5 nm; for particles having an average diameter between about 1 μm and about 20 μm, the coating thickness should average between about 1 nm and about 10 nm; for particles having an average diameter between about 20 μm and about 700 μm, the coating thickness should average between about 1 nm and about 100 nm.

[0141] We have found that applying a coating / shell followed by one or more deagglomeration steps (such as sonication) can create abrasions, pinholes, breaks, gaps, cracks and / or voids (hereinafter referred to as 'cracks') in the layer / coating, as the coated particles are essentially 'bound' or 'glued' together more tightly directly after the thicker coating is applied. Once deagglomeration occurs, this may expose the core containing the biologically active ingredient to the elements.

[0142] As described in International Patent Application PCT / GB2020 / 053129, we surprisingly found that performing a mechanical screening method (by hand or by manually forcing the particles through a sieve as opposed to the ultrasonic treatment described in International Patent Application WO 2014 / 187995, or the mechanical screening method mentioned in International Patent Application PCT / GB2020 / 053129) can produce significantly fewer pinholes, gaps or cracks in the coating material. In turn, the following particles are produced: the particles are not only completely covered by the layer / coating, but also covered in a way that allows the particles to be easily deagglomerated (for example using non-aggressive techniques such as vortexing), so as not to destroy the layer of coating material that has been formed before and / or during drug formulation.

[0143] Very surprisingly, we also found that when using the vibratory screening technique, the low frequency of pinholes, gaps or cracks mentioned above in the coating material can be maintained. This is surprising because the technology described herein uses a stainless steel sieve (rather than the softer polymer sieve used in the mechanical screening method mentioned in International Patent Application PCT / GB2020 / 053129), which may be preferred for such vibratory screening techniques. Previous attempts to manually force particles through a metal sieve resulted in significant formation of pinholes, gaps or cracks in the coating material.

[0144] For example, if it is intended to provide a sample in suspension prior to administration to a patient, it is therefore desirable to provide deagglomerated primary particles without pinholes or cracks in the coating. Such cracks would lead to an undesirable initial peak (burst release) in the plasma concentration of the active ingredient immediately after administration.

[0145] The methods described herein result in deagglomerated coated particles that are substantially free of such cracks through which the active ingredient can be released in an uncontrolled manner. By 'substantially free of such cracks' in the coating, we mean that less than about 1% of the surface of the coated particles includes abrasions, pinholes, breaks, gaps, cracks and / or voids through which the active ingredient could potentially be exposed (e.g., to the elements).

[0146] In this regard, the (e.g. inorganic, such as mixed oxide) coating typically completely surrounds, encloses and / or encapsulates the solid core. In this way, the risk of an initial drug concentration burst due to direct contact of the drug with a solvent in which the relevant active ingredient is soluble is minimized. This may include not only body fluids, but also any medium in which such coated particles may be suspended prior to injection.

[0147] Therefore, in one embodiment of the present invention, there is provided a particle as disclosed above, wherein the coating surrounding, enclosing and / or encapsulating the core covers at least about 50%, such as at least about 65%, including at least about 75%, such as at least about 80%, more specifically at least about 90%, such as at least about 91%, such as at least about 92%, such as at least about 93%, such as at least about 94%, such as at least about 95%, such as at least about 96%, such as at least about 97%, such as at least about 98%, such as at least about 99%, such as about or about 100% of the surface of the solid core, so that the coating substantially completely surrounds, surrounds and / or encapsulates the core.

[0148] As used herein, the term 'the coating substantially completely surrounds, encloses and / or encapsulates the core' means covering at least about 98%, or at least about 99%, of the surface of the solid core.

[0149] Although some minor cracks may appear in the coating without affecting its basic function in controlled release, in another embodiment, particles as disclosed above are provided, wherein at least about 90% of the particles do not have cracks in the coating surrounding, enclosing 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 about 100% of the particles do not have the cracks.

[0150] In summary, the coating material layer may have a substantially uniform thickness over the surface area of ​​the particles. By "substantially uniform" thickness, we mean that at least about 10%, such as about 25%, for example about 50% of the coated particles present in the composition prepared by the method of the present invention have a thickness that varies by no more than about ±20%, including ±50%, of the average thickness as measured by TEM.

[0151] Coating materials that may be applied to the core may be pharmaceutically acceptable in that they should be substantially non-toxic.

[0152] The coating material may comprise an organic material or a polymeric material, such as polyamide, polyimide, polyurea, polyurethane, polythiourea, polyester or polyimide. The coating material may also comprise a mixed material (such as between an organic material and an inorganic material) comprising a material as a combination of 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.

[0153] The inorganic coating material may comprise one or more metals or metalloids, or may comprise one or more metal- or metalloid-containing compounds, such as metal or metalloid oxides, metal or metalloid nitrides, metal or metalloid sulfides, metal or metalloid selenides, metal or metalloid carbonates and / or other ternary compounds, etc. Preferred are metal and metalloid hydroxides, and especially metal and metalloid oxides, especially metal oxides.

[0154] 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.

[0155] The metal-containing or metalloid-containing precursor can be applied as the first reactant gas or the second reactant gas during the vapor deposition technique. In some methods, it may be advantageous to start with a metal-containing or metalloid-containing precursor, while in other methods, it may be advantageous to start with an oxygen precursor (such as water). This may depend on the moiety on the surface of the compound to be coated.

[0156] In embodiments of the present invention, each iteration of applying the vapor deposition technique may be performed using the same or different first reactant gas and second reactant gas as the previous iteration of step (b). For example, the metal-containing or metalloid-containing precursor used in one iteration may be different from the metal-containing or metalloid-containing precursor used in a subsequent iteration. Further, in embodiments of the present invention involving particle deagglomeration between iterations of the vapor deposition technique, the metal-containing or metalloid-containing precursor may alternate from the first reactant gas to the second reactant gas, or vice versa.

[0157] As mentioned above, since the composition prepared by the method of the present invention may comprise two or more discrete layers of inorganic coating material, the nature and chemical composition of those layers may vary from layer to layer.

[0158] The various layers may also comprise mixtures of two or more inorganic materials, such as metal oxides or metalloid oxides, and / or may comprise multiple layers or combinations of different inorganic or organic materials to modify the properties of the layer.

[0159] Coating materials that may be mentioned include those comprising aluminum oxide (Al2O3), titanium dioxide (TiO2), iron oxide (FexOy, such as 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.

[0160] The coating material layer in the composition prepared by the method of the present invention (on an individual or collective basis) can be essentially composed of (e.g., greater than about 80%, such as greater than about 90%, for example, about 95%, such as about 98%) iron oxide, titanium dioxide, or more preferably zinc oxide, silicon oxide and / or aluminum oxide.

[0161] The method of the invention is particularly useful when the coating material applied to the core comprises zinc oxide, silicon dioxide and / or aluminum oxide.

[0162] It is further preferred that the inorganic coating material comprises a mixture of:

[0163] (i) zinc oxide (ZnO); and

[0164] (ii) one or more other metal and / or metalloid oxides,

[0165] wherein the atomic ratio ((i):(ii)) is between at least about 1:6 and up to and including about 6:1.

[0166] A coating comprising a mixture of zinc oxide and one or more other metal oxides and / or metalloid oxides is hereinafter referred to as a 'mixed oxide' coating or coating material.

[0167] Thus, the core containing the biologically active agent can be coated with a coating material comprising a mixture of zinc oxide and one or more other metal oxides and / or metalloid oxides, wherein the atomic ratio of zinc oxide to other oxides is: i.e., at least about 1:1 (e.g., at least about 1.5:1, such as at least about 2:1), including at least about 2.25:1, such as at least about 2.5:1 (e.g., at least about 3.25:1 or at least about 2.75:1 (including 3:1)), and up to (i.e., not exceeding) and including about 6:1, including up to about 5.5:1, or up to about 5:1, such as up to about 4.5:1, including up to about 4:1 (e.g., up to about 3.75:1), or vice versa.

[0168] To prepare a mixed oxide coating having an atomic ratio of zinc oxide to the one or more other metal and / or metalloid oxides of about 1:1 up to and including about 6:1, the skilled artisan will appreciate that for each ALD cycle (i.e., monolayer) of the other oxide, between about 1 and about 6 ALD cycles of zinc oxide must also be deposited. For example, for a mixed oxide coating to be formed with an atomic ratio of 3:1 (zinc: other oxide), three zinc-containing precursor pulses may each be followed by a second precursor pulse to form three zinc oxide monolayers, which may then be followed by one precursor pulse containing another metal and / or metalloid, followed by the second precursor pulse to form one monolayer of the oxide of the other metal and / or metalloid. Alternatively, six zinc oxide monolayers may be followed by two other oxide monolayers or any other combination to provide an overall atomic ratio of about 3:1. In this regard, the order of the pulses used to produce the relevant oxides is not critical, provided that the resulting atomic ratio is within the relevant range.

[0169] Other coating materials, such as pharmaceutically acceptable and substantially non-toxic coating materials, may also be applied between separate coatings as described herein (e.g., between separate deagglomeration steps) and / or at the same time as the coatings. Such materials may comprise multiple layers of the mixed oxide and one or more different inorganic or organic materials or a combination of the mixed oxide and one or more different inorganic or organic materials to modify the properties of the layers.

[0170] In ALD, the coating material layer can be applied at a process temperature of about 20°C to about 800°C, or about 40°C to about 200°C, for example about 40°C to about 150°C, such as about 50°C to about 100°C. The optimal process temperature depends on the reactivity of the precursors and / or substances (including bioactive agents) used in the core and / or the melting point of the core material. When the core to be coated contains a bio-active ingredient, it is preferred to use a lower temperature, such as about 20°C to about 100°C. Specifically, in one embodiment of the method, a temperature of about 20°C to about 80°C is used, such as about 30°C to about 70°C, such as about 40°C to about 60°C, such as about 50°C.

[0171] We have found that when ALD is used to apply coatings comprising zinc oxide at lower temperatures, such as about 50°C to about 100°C (unlike other coating materials such as aluminum oxide and titanium oxide that form amorphous layers), the coating material is predominantly crystalline in nature.

[0172] Without being limited by theory, since zinc oxide is crystalline, if only zinc oxide is used as the coating material, it is understood that interfaces may form between adjacent zinc oxide crystals deposited by ALD, through which a carrier system, medium or solvent (e.g., an aqueous solvent system) in which zinc oxide is partially soluble may enter after being suspended therein. It is believed that this may result in dissolution that is too rapid for the storage composition it is intended to prepare.

[0173] We have now found that these problems can be alleviated by preparing mixed oxide coatings as described herein. In particular, we have now found that these problems can be alleviated by preparing a mixture of two or more metal oxide 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), we have been able to coat the active ingredient with a coating that appears to be essentially amorphous, or a composition between a crystalline material and an amorphous material, and / or in which the ingress of injection vehicles (such as water) can be reduced. In this regard, it seems to us that by employing a mixed oxide according to this aspect of the invention, the presence of the aforementioned perceived interface can be reduced or completely avoided, either in a heterogeneous manner (where another oxide is 'filling' the gap formed by the interface) or in a homogeneous manner (where a true composition of mixed oxide material is formed during deposition in a manner that potentially avoids the interface in the first place).

[0174] Thus, a method for preparing a plurality of coated particles according to the invention is provided, wherein the coated particles are prepared by applying at least two metal oxide and / or metalloid oxide precursors forming a mixed oxide on a solid core and / or a previously coated solid core by a vapor deposition technique. The precursors for forming the metal oxide or metalloid oxide typically include an oxygen precursor, such as water, oxygen, ozone and / or hydrogen peroxide; and a metal and / or metalloid compound, typically an organometallic compound or an organometalloid compound.

[0175] Non-limiting examples of precursors are as follows: The precursor of zinc oxide can be water and di-C1-C5 alkyl zinc, such as diethyl zinc. The precursor of aluminum oxide can be water and tri-C1-C5 alkyl aluminum, such as trimethyl aluminum. The precursor of silicon oxide (silicon dioxide) can be water as an oxygen precursor, as well as silane, alkyl silane, aminosilane and tetraethoxysilane. The precursor of iron oxide includes oxygen, ozone and water as an oxygen precursor; and di-C1-C5 alkyl iron, dicyclopropyl iron and FeCl3. It should be understood that those skilled in the art understand what precursors are suitable for the purposes disclosed herein.

[0176] Although the plurality of coated particles produced according to the method of the present invention are substantially free of the aforementioned cracks in the applied coating through which the active ingredient could potentially be exposed (e.g., to the natural environment), two further optional steps may be applied to the plurality of coated particles before further pharmaceutical formulation processing thereof.

[0177] A first optional step may include, after the final deagglomeration step as described above, applying a final outer coating layer, the thickness of this outer "overcoating" layer / coating, or "sealing shell" (these terms are used interchangeably herein) necessarily being thinner than the individual layers / coatings / shells (or "sub-shells") previously applied.

[0178] Thus, the thickness may average no more than about 0.7 times (e.g., about 0.6 times) the thickness of the widest subshell previously applied. Alternatively, the thickness may average no more than about 0.7 times (e.g., about 0.6 times) the thickness of the last subshell applied, and / or may average no more than about 0.7 times (e.g., about 0.6 times) the average thickness of all previously applied subshells. For particles up to about 20 μm, the thickness may average in the region of about 0.3 nm to about 10 nm. For larger particles, the thickness may average no more than about 1 / 1000 of the average diameter of the coated particles based on weight, number, or volume.

[0179] The role of a sealing shell such as this is to provide a 'sealing' outer coating layer on the particle which overlies those cracks, thereby producing a particle that is not only completely covered by the sealing shell, but is also covered in a manner that enables the particle to be easily deagglomerated (e.g. using non-aggressive techniques such as vortexing) without disrupting the sub-shell that has been formed underneath prior to and / or during drug formulation.

[0180] For the reasons described herein, it is preferred that the sealed shell does not contain zinc oxide. On the other hand, the sealed shell may contain silicon dioxide, or more preferably aluminum oxide.

[0181] A second optional step may include ensuring that the few remaining particles with broken and / or cracked shells / coatings are subjected to a treatment in which all of the particles are suspended in a solvent in which the active ingredient is soluble (e.g., its solubility is at least about 1 mg / mL), but the least soluble material in the coating is insoluble (e.g., its solubility does not exceed about 0.1 μg / mL), and then separating the solid material particles from the solvent by, for example, centrifugation, sedimentation, flocculation and / or filtration, thereby resulting in substantially intact particles remaining.

[0182] The above mentioned optional steps provide a means to potentially further reduce the likelihood of a (potentially) undesirable initial peak (burst) in the plasma concentration of the active ingredient, as discussed above.

[0183] At the end of the process, the coated particles may be dried using one or more of the techniques described above for drying the cores. Drying may be carried out in the absence or presence of one or more pharmaceutically acceptable excipients (eg, sugars or sugar alcohols).

[0184] Alternatively, at the end of the process, the separated particles may be resuspended in a solvent (eg water, in the presence or absence of one or more pharmaceutically acceptable excipients as defined herein) for subsequent storage and / or administration to a patient.

[0185] Prior to applying the first layer of coating material or between successive coats, the core and / or partially coated particles may undergo one or more alternative and / or preparatory surface treatments. In this regard, one or more intermediate layers comprising different materials (i.e. in addition to inorganic materials) may be applied to the relevant surfaces, for example to protect the core or partially coated particles from undesirable reactions with precursors during the coating step / deposition process, to improve coating efficiency or to reduce agglomeration.

[0186] The intermediate layer may, for example, comprise one or more surfactants, the purpose of which is to reduce the aggregation of the particles to be coated and to 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, for example Tween 80. Alternatively, if the active ingredient used as part of the core (or used as the core) is susceptible to reacting with one or more precursor compounds that may be present in the gas phase during the coating (e.g. ALD) process, the core may undergo a preliminary surface treatment.

[0187] Application of an 'intermediate' layer / surface treatment of this nature may alternatively be achieved by means of liquid phase non-coating techniques followed by freeze drying, spray drying or other drying methods to provide particles with a surface layer to which a coating material may then be applied.

[0188] The outer surface of the particles of the composition prepared by the method of the present invention can also be derivatized or functionalized, for example, by attaching one or more chemical compounds or moieties to the outer surface of the final layer of a coating material, which coating material, for example, has a compound or moiety that enhances the targeted delivery of the particles in a patient administering the nanoparticles. Such compounds can be organic molecules (such as PEG) polymers, antibodies or antibody fragments, or receptor binding proteins or peptides, etc.

[0189] Alternatively, the moiety can be an anchoring group, such as a moiety comprising a silane functional group (see, e.g., Herrera et al., J. Mater. Chem., 18, 3650 (2008) and US 8,097,742). Another compound, such as a desired targeting compound, can be attached to such an anchoring group by means of covalent bonding or non-covalent bonding (including hydrogen bonding or van der Waals bonding) or a combination thereof.

[0190] The presence of such anchoring groups can provide a versatile tool for targeted delivery to specific sites in the body. Alternatively, the use of compounds such as PEG may result in particles circulating in the bloodstream for a longer duration, thereby ensuring that they do not accumulate in the liver or spleen (the body's natural mechanism for eliminating particles, which can prevent delivery to diseased tissues).

[0191] The compositions prepared by the methods of the present invention are suitable for administration to a patient at the time of preparation of the composition (i.e. as a plurality of particles) or are preferably formulated together with one or more pharmaceutically acceptable excipients, including adjuvants, diluents or carriers, for use in the medical or veterinary fields (including use in therapy, and / or use in diagnostics if the core contains diagnostic material) .

[0192] Further provided are compositions prepared by the methods of the present invention for use in medicine, diagnostics and / or in veterinary practice, as well as pharmaceutical (or veterinary) preparations comprising compositions prepared by the methods of the present invention and pharmaceutically (or veterinarily) acceptable adjuvants, diluents or carriers.

[0193] The compositions prepared by the methods of the invention can be administered topically, topically or systemically, for example orally (enterally), by injection or infusion, intravenously or intraarterially (including by intravascular or other perivascular devices / dosage forms (e.g., stents)), intramuscularly, intraosseously, intracerebrally, intracerebroventricularly, intrasynovially, intrasternally, intrathecally, intralesionally, intracranially, intratumorally, cutaneously, intradermally, subcutaneously, transmucosally (e.g., sublingually or buccally), rectally, transdermally, nasally, pulmonary (e.g., by inhalation, tracheally or bronchially), topically, or by any other parenteral route, such as subcutaneously or intramuscularly, optionally in the form of a pharmaceutical (or veterinary) formulation comprising the compound in a pharmaceutically (or veterinarily) acceptable dosage form.

[0194] Incorporation of the composition prepared by the method of the present invention into a pharmaceutical formulation can be achieved with due consideration of the intended route of administration and standard pharmaceutical practice. Pharmaceutically acceptable excipients (such as carriers) can be chemically inert to the bioactive agent and may not have harmful side effects or toxicity under the conditions of use. Such pharmaceutically acceptable carriers can also impart immediate release or modified release of the active agent from the particles in the composition prepared by the method of the present invention.

[0195] The pharmaceutical (or veterinary) preparation comprising the composition prepared by the method of the present invention may comprise different types of particles, for example particles comprising different active ingredients, particles comprising different functionalizations (as described above), particles of different sizes and / or different thicknesses of coating material layers, or combinations thereof. By combining particles having different coating thicknesses and / or different core sizes in a single pharmaceutical preparation, the release of the drug after administration to a patient can be controlled (e.g., altered or prolonged) over a specific period of time.

[0196] For oral administration (i.e., administration to the gastrointestinal tract through the mouth in a swallowing manner), the composition prepared by the method of the present invention can be formulated into a variety of dosage forms. Pharmaceutically acceptable carriers or diluents can be solid or liquid. Solid preparations include granules (wherein the granules may contain some or all of the various particles of the composition prepared by the method of the present invention in the presence of, for example, carriers and other excipients (such as binders or pH adjusters), compressed tablets, pills, lozenges, capsules, cachets, and the like. Carriers include materials well known to those skilled in the art, including those disclosed above in connection with the formulation of the bioactive agent in the core, as well as magnesium carbonate, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoa butter, lactose, microcrystalline cellulose, low crystalline cellulose, and the like.

[0197] Solid dosage forms may include other excipients, such as flavoring agents, lubricants, binders, preservatives, disintegrants and / or encapsulating materials. For example, the compositions prepared by the method of the present invention may be encapsulated in, for example, soft shell capsules or hard shell capsules (e.g., gelatin capsules).

[0198] Compositions prepared by the methods of the present invention and formulated for rectal administration may include suppositories which may contain, for example, suitable non-irritating excipients such as cocoa butter, synthetic glycerides or polyethylene glycols which are solid at ordinary temperatures but liquefy and / or dissolve in the rectal cavity to release particles of the composition prepared by the methods of the present invention.

[0199] For parenteral administration, such as subcutaneous injection and / or intramuscular injection, the composition prepared by the method of the present invention may be in the form of sterile injection and / or infusion dosage forms, for example, sterile aqueous or oily suspensions of the composition prepared by the method of the present invention.

[0200] Sterile aqueous suspensions of particles of the composition prepared by the method of the present invention can be formulated according to techniques known in the art. The aqueous medium should contain at least about 50% water, but may also contain other aqueous excipients, such as Ringer's solution, and may also contain polar cosolvents (e.g., ethanol, glycerol, propylene glycol, 1,3-butylene glycol, various molecular weight polyethylene glycols and tetraethylene glycol); viscosity increasing agents or thickening agents (e.g., carboxymethylcellulose, microcrystalline cellulose, hydroxypropyl methylcellulose, hydroxyethylcellulose, ethyl hydroxyethylcellulose, sodium starch glycolate, poloxamers such as poloxamer 407, polyvinylpyrrolidone, cyclodextrins such as hydroxypropyl-β-cyclodextrin, polyvinylpyrrolidone and various molecular weight polyethylene glycols); surfactants / wetting agents to obtain a uniform suspension (e.g., sorbitan esters, sodium lauryl sulfate; monoglycerides, polyoxyethylene esters, polyoxyethylene alkyl ethers, polyoxyglycerol esters and preferably Tweens (polysorbates), such as Tween 80 and Tween 20). Preferred ingredients include isotonicity adjusting agents (e.g., sodium lactate, dextrose, and especially sodium chloride); pH adjusting agents and / or buffers (e.g., citric acid, sodium citrate, and especially phosphate buffers, such as disodium hydrogen phosphate dihydrate, sodium acid phosphate, sodium dihydrogen phosphate monohydrate, and combinations thereof, which may be used in conjunction with standard inorganic acids and bases such as hydrochloric acid and sodium hydroxide); and other ingredients, such as mannitol, croscarmellose sodium, and hyaluronic acid.

[0201] The oily or oil-based carrier system may contain one or more pharmaceutically or veterinarily acceptable liquid lipids, which may include fixed oils such as mono-, di- or triglycerides including miglyol (e.g. 812N), propylene glycol dicaprylic decanoate (Miglyol 840, C8 / C10 ester), tricaprylin (Miglyol oil), gelucire 43 / 01, kollisolv GTA, labrafil. The carrier system 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 pharmaceutically acceptable oils, such as olive oil, peanut oil, soybean oil, corn oil, cottonseed oil, sesame oil, castor oil, oleic acid and their polyoxyethylated versions (e.g., sorbitan trioleate, propylene glycol monolaurate 90, capryol PGMC, PEG-60 hydrogenated castor oil, polyoxyethylene 35 castor oil). More preferred carrier systems include monoglycerides, diglycerides and / or triglycerides, with medium chain triglycerides, such as alkyl chain triglycerides (e.g., C6-C12 alkyl chain triglycerides) being the most preferred.

[0202] Such injection suspensions can be formulated according to techniques well known to those skilled in the art by using suitable dispersing agents or wetting agents (for example, Tweens such as Tween 80) and suspending agents.

[0203] The composition suitable for injection prepared by the method of the present invention may be in the form of a liquid, sol, paste or gel, which is administrable via a surgical administration device (eg, a syringe with a needle for injection, a catheter, etc.) forming a depot preparation.

[0204] Use of the compositions prepared by the process of the present invention can control the dissolution rate and pharmacokinetic characteristics by reducing any burst effect as defined above and / or by reducing the Cmax in the plasma concentration-time curve and thereby increase the length of release of the bioactive ingredient from the formulation.

[0205] These factors not only reduce the frequency with which the formulations obtained by the methods of the present invention need to be administered to the subject, but also allow the subject to spend more time as an outpatient and therefore have a better quality of life.

[0206] The compositions prepared by the method of the present invention also have the advantage of providing lower daily exposure to potentially toxic drugs by controlling the release of the active ingredient at a steady rate over an extended period of time, which is expected to reduce unwanted side effects.

[0207] The composition prepared by the process of the present invention may be contained in a reservoir as well as an injection device or an infusion device, wherein the coated particles and the carrier system are contained separately and wherein the admixture occurs before and / or during injection or infusion.

[0208] The compositions prepared by the methods of the present invention can also be formulated for inhalation, e.g., as inhalation powders for use with dry powder inhalers (see, e.g., those described by Kumaresan et al., Pharma Times, 44, 14 (2012) and Mack et al., Inhalation, 6, 16 (2012)), the relevant disclosures of which are hereby incorporated herein by reference. Suitable particle sizes of the various particles in the compositions prepared by the methods of the present invention for inhalation into the lungs are in the range of about 2 μm to about 10 μm.

[0209] Compositions prepared by the method of the present invention can also be formulated for external application to skin or to mucosa.For external application, pharmaceutical preparations can be provided in the form of, for example, lotions, gels, pastes, tinctures, transdermal patches, gels for delivery through mucosa, and all of these can comprise compositions prepared by the method of the present invention. Said composition can also be prepared with suitable ointment, and this ointment contains the composition prepared by the method of the present invention suspended in a carrier, and this carrier is such as mineral oil, liquid vaseline, white vaseline, propylene glycol, polyoxyethylene polyoxypropylene compounds, emulsifying wax or water. Suitable carriers for lotions or creams include mineral oil, sorbitan monostearate, polysorbate 60, hexadecyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

[0210] The pharmaceutical formulation may comprise between about 1% to about 99%, such as between about 10% (such as about 20%, e.g. about 50%) to about 90% by weight of coated particles, with the remainder being made up by the carrier system and / or other pharmaceutically acceptable excipients.

[0211] The pharmaceutical preparation may be in the form of a liquid, sol or gel, which is administrable via a surgical administration device (eg, needle, catheter, etc.) forming a depot preparation.

[0212] In any case, the composition prepared by the method of the present invention can be formulated with conventional pharmaceutical additives and / or excipients used in the art for preparing pharmaceutical preparations, and then incorporated into various pharmaceutical preparations and / or dosage forms using standard techniques (see, e.g., Lachman et al., 'The Theory and Practice of Industrial Pharmacy', Lea & Febiger, 3rd edition (1986); 'Remington: The Science and Practice of Pharmacy', Troy (ed.), University of the Sciences in Philadelphia, 21st edition (2006); and / or 'Aulton's Pharmaceutics: The Design and Manufacture of Medicines', Aulton and Taylor (ed.), Elsevier, 4th edition, 2013), and the documents cited therein, all of which are incorporated herein by reference in their relevant disclosures. Otherwise, the preparation of suitable preparations can be achieved non-inventively by a skilled person using conventional techniques.

[0213] According to a further aspect of the present invention, there is provided a method for preparing a pharmaceutical or veterinary formulation, the method comprising mixing the coated particles prepared as described herein with a pharmaceutically or veterinarily acceptable adjuvant, diluent or carrier.

[0214] Preferably, such formulations are for injection and / or infusion and therefore comprise one or more compositions prepared by the method of the present invention suspended in a pharmaceutically or veterinarily acceptable aqueous and / or oily carrier.

[0215] Further provided are injectable and / or infusible dosage forms comprising a composition prepared by the method of the present invention, wherein the composition is contained in a reservoir connected and / or associated with an injection device or infusion device (e.g., a syringe with a needle for injection, a catheter, etc.). In this regard, the composition prepared by the method of the present invention can be stored before being loaded into a suitable injectable and / or infusible administration device (e.g., a syringe with a needle for injection), or can even be prepared immediately before being loaded into such an administration device.

[0216] There is therefore further provided a kit of parts comprising:

[0217] (a) a composition prepared by the method of the present invention; and

[0218] (b) a pharmaceutically acceptable or veterinarily acceptable carrier system,

[0219] and kits of parts comprising the compositions prepared by the methods of the invention together with instructions for the end user to mix the particles with a pharmaceutically or veterinarily acceptable aqueous and / or oily carrier system.

[0220] Also provided is a preloaded injectable and / or infusible dosage form as described above, but modified by comprising at least two chambers, one of the at least two chambers having a composition prepared by the method of the present invention disposed therein, and another of the at least two chambers having a pharmaceutically acceptable or veterinarily acceptable carrier system disposed therein, wherein admixture, suspension creation or otherwise occurs prior to and / or during injection or infusion.

[0221] Wherever the word "about" is used herein, for example in the context of an amount (e.g., concentration, dimension (size and / or weight), time period), relative amount (percentage, weight ratio, atomic ratio, dimension ratio, aspect ratio, ratio, factor or fraction), relative humidity, lux, temperature or pressure, it is understood that such variables are approximate and thus may vary by ±15%, such as ±10%, for example ±5%, and preferably ±2% (e.g., ±1%) compared to the numbers specified herein. This is true even if such numbers are initially presented as percentages (e.g., 'about 15%' may mean about ±15% of the number 10, which is any number between 8.5% and 11.5%).

[0222] The compositions prepared by the methods of the present invention allow for the formulation of a variety of pharmaceutically active compounds. The compositions prepared by the methods of the present invention can be used to effectively treat a variety of conditions, depending on the biologically active agent included.

[0223] The composition prepared by the method of the present invention can be further formulated in the form of an injectable suspension of coated particles, the size distribution of the coated particles is both uniform and capable of forming a stable suspension (i.e., no sedimentation) in the injection liquid, and can be injected through a needle. In this aspect, the composition prepared by the method of the present invention can include an aqueous medium, the aqueous medium includes an inactive ingredient that can prevent premature gelation of the composition prepared by the method of the present invention, and or the viscosity of the aqueous medium is sufficient to prevent sedimentation, which produces a non-'homogeneous' suspension and thus creates a risk of underdosing or overdosing of the active ingredient.

[0224] Furthermore, the composition prepared by the method of the present invention can be stored under normal storage conditions and maintain its physical and / or chemical integrity.

[0225] The phrase 'maintaining physical and chemical integrity' essentially means chemical stability and physical stability.

[0226] By 'chemical stability' we define any composition prepared by the method of the present invention as being able to be stored under normal storage conditions (with or without appropriate pharmaceutical packaging) without undergoing a significant degree of chemical degradation or decomposition.

[0227] By 'physical stability', we define any composition prepared by the method of the present invention as being able to be stored under normal storage conditions (with or without appropriate pharmaceutical packaging) without undergoing a significant degree of physical transformation, such as sedimentation as described above, or changes in the properties and / or integrity of the coated particles, such as changes in the coating itself or in the active ingredient (including dissolution, solvation, solid-state phase transition, etc.).

[0228] Examples of 'normal storage conditions' for the compositions prepared by the method of the invention include a temperature between about -50°C and about +80°C (preferably between about -25°C and about +75°C, such as about 50°C), and / or a pressure between about 0.1 bar and about 2 bar (preferably atmospheric pressure), and / or exposure to UV / visible light of about 460 lux, and / or a relative humidity between about 5% and about 95% (preferably about 10% to about 40%), for an extended period of time (i.e. greater than or equal to about twelve months, such as about six months).

[0229] Under such conditions, it can be found that the composition prepared by the method of the present invention undergoes chemical and / or physical degradation / decomposition in appropriate cases in an amount less than about 15%, more preferably less than about 10%, and especially less than about 5%. The skilled person will understand that the upper and lower limits of temperature and pressure mentioned above represent extremes of normal storage conditions, and that certain combinations of these extremes (e.g., a temperature of 50° C. and a pressure of 0.1 bar) will not be experienced during normal storage.

[0230] Furthermore, the compositions prepared by the methods of the present invention may provide a release and / or pharmacokinetic profile that minimizes any burst effect and / or minimizes Cmax, characterized by a concentration maximum shortly after administration.

[0231] The compositions and methods described herein may have the following advantages: they may be more convenient for physicians and / or patients to treat relevant conditions with a particular biologically active agent, be more effective, be less toxic, have a broader spectrum of activity, be more potent, produce fewer side effects, or may have other useful pharmacological properties, compared to any similar treatment described in the prior art for the same active ingredient.

[0232] The present invention is illustrated by the following examples and with reference to the accompanying drawings, but is not limited in any way to the present invention, in which Figures 1 to 4 Dose-adjusted plasma concentration versus time curves after administration of samples prepared according to the corresponding Examples are shown.

[0233] Examples

[0234] Comparative Example 1

[0235] Coating of Lenalidomide Microparticles Using Continuous Flow ALD I

[0236] Lenalidomide micron particles 99.9% (APIChem, China) were used as received. The average diameter of the lenalidomide particles was 10 μm, as determined by laser diffraction (Shimadzu, SALD-7500nano, Kyoto, Japan). The particle size distribution determined by laser diffraction was as follows: 10 2.7μm; D 50 10.4μm and D 90 24.9μm.

[0237] Microparticles or solid cores are coated as described in steps 1 to 4.

[0238] 1. Load the micron particles into the ALD reactor (Picosun, SUNALE TM R series, Espoo, Finland), and three ALD cycles using diethylzinc and water as precursors were performed at a reactor temperature of 50°C, followed by one ALD cycle with trimethylaluminum and water as precursors. This was repeated six times, i.e., 6x (3 Zn cycles + 1 Al cycle), resulting in a total of 24 cycles. By this method, a first layer of a mixed oxide having a zinc:aluminum atomic ratio of approximately 3:1 was formed.

[0239] The ALD reactor includes a reaction chamber into which the micron particles are loaded. The ALD reactor further includes precursor bottles that independently contain each precursor, each precursor bottle being connected to the reaction chamber via a valve. The ALD reactor also includes a pump and associated piping for pumping an inert gas, such as nitrogen, through the reaction chamber, the pump also being connected to the reaction chamber via a valve.

[0240] The ALD cycle is performed as follows, where steps a to e represent the first cycle, subsequent cycles begin with step d as specified in step 1, and the final cycle ends at step k:

[0241] a. Reagent pulse: Open the valve of the precursor bottle for 0.1s, evaporate the water and bring it into the reaction chamber by inert nitrogen gas. The water is adsorbed onto the surface of the drug particles, presenting hydroxyl groups on the outside of the particles.

[0242] b. The reactor was then pumped for 3 s.

[0243] c. Repeat steps a to b above 100 times.

[0244] d. Purge pulse: Purge the chamber with nitrogen. Gaseous water and organic gases (in case this is not the first cycle) will be removed.

[0245] e. Reagent pulse: The valve of the precursor bottle is opened for 0.1 s, diethylzinc and trimethylaluminum are evaporated and carried into the reaction chamber by inert nitrogen. Dimethylzinc or trimethylaluminum is adsorbed onto the surface of the drug particles and reacts with the hydroxyl groups. This releases ethane from diethylzinc or methane from trimethylaluminum.

[0246] f. The reactor was then pumped for 3 s.

[0247] g. Repeat steps e to f above 100 times.

[0248] h. Purge pulse: Purge the chamber with nitrogen to remove unreacted reagents and organic gases.

[0249] i. Reagent pulse: The valve of the precursor bottle is opened for 0.1 s, and water is evaporated and carried into the reaction chamber by inert nitrogen. Water is adsorbed to the surface of the drug particles and reacts with the metal organic surface. The remaining ethyl or methyl groups are converted to ethane and methane, respectively. As a result, the surface is covered with a metal oxide layer, presenting hydroxyl groups on the outside of the particles.

[0250] j. After this the reactor was pumped for 3 s.

[0251] k. Repeat the above steps i to j 100 times

[0252] l. Repeat the cycle starting from step d.

[0253] 2. Next, the powder was taken out from the reactor and deagglomerated by a sonic sieving machine (Tsutsui Sonic Sonic Agitation Sieving Machine SW-20AT) with a 32 μm mesh size sieve.

[0254] 3. The resulting deagglomerated powder was reloaded into the ALD reactor and steps 1 to 2 were repeated twice to form a second and third layer of a mixed oxide having an approximately 3:1 zinc:aluminum atomic ratio.

[0255] 4. The resulting deagglomerated powder was reloaded into the ALD reactor and step 1 was repeated once.

[0256] Comparative Example 2

[0257] Coating of Lenalidomide Microparticles Using Continuous Flow ALD II

[0258] This is similar to Comparative Example 1, but two additional layers are formed. In other words, step 3 is repeated once before proceeding to step 4.

[0259] Example 3

[0260] Coating of Lenalidomide Microparticles Using Stopped-Flow ALD I

[0261] Lenalidomide micronized particles 99.9% (Kyongbo, South Korea) were used as received. The average diameter of the lenalidomide particles was 4 μm, as described in Comparative Example 1 above. The particle size distribution determined by laser diffraction was as follows: 10 0.6μm; D 50 3.9μm and D 90 15.6μm.

[0262] The microparticles were coated essentially as described in steps 1 to 4 of Comparative Example 1, except that the ALD cycles were performed as follows (steps a to d represent the first cycle, with subsequent cycles starting from step a as specified in step e).

[0263] a. Reagent pulse 1:

[0264] i. Close the valve on the pipe between the pump and the ALD reactor.

[0265] ii. Open the valve on the water precursor bottle for 1 s to allow the evaporated water to fill the reaction chamber.

[0266] iii. The valve of the water precursor bottle was closed and the chamber was left to stand for 30 s (soak time) before the pump valve was opened again to ensure that the water vapor was adsorbed to the surface of the drug particles, presenting hydroxyl groups on the outside or particles. iv. The reactor was then pumped for 9 s.

[0267] v. Repeat steps i to iv above 20 times.

[0268] b. Purge pulse: Purge the chamber with nitrogen in a continuous flow. Gaseous water and organic gases (in case this is not the first cycle) will be removed.

[0269] c. Reagent pulse:

[0270] i. Close the valve on the pipe between the pump and the ALD reactor.

[0271] ii. The valve on the diethylzinc or trimethylaluminum precursor bottle was opened for 1 second to allow the evaporated metal-containing precursor to fill the reaction chamber.

[0272] iii. The valve of the precursor bottle was closed and the chamber was left to stand for 30 s (soak time) before opening it to the pump again to ensure that the metal-containing precursor vapor reacted with the hydroxyl groups on the surface of the drug particles.

[0273] iv. The reactor was then pumped for 9 s.

[0274] v. Repeat steps i-iv above 20 times.

[0275] d. Purge pulse: Purge the chamber with nitrogen in a continuous flow to remove unreacted reagents and organic gases.

[0276] e. Repeat the cycle starting from step a.

[0277] Example 4

[0278] Coating of Lenalidomide Microparticles Using Stopped-Flow ALD II

[0279] This is similar to Comparative Example 3, but two additional layers are formed. In other words, step 3 is repeated once before proceeding to step 4.

[0280] Example 5

[0281] Determination of drug loading

[0282] This example describes the analysis of the lenalidomide drug loading of the particles obtained in Comparative Examples 1 and 2 and Examples 3 and 4.

[0283] Materials and methods

[0284] Drug loading (i.e., w / w % of lenalidomide in powder) was determined using UPLC using a Prominence-i (Shimadzu, Japan) equipped with a diode array detector (Shimadzu, Japan) set at a wavelength of 223 nm, using a 4.6×100 mm, 2.6 μm particle, C18WP column (SunShell, ChromaNik Technologies Inc, Osaka, Japan). The material was dissolved in 2 M phosphoric acid in acetonitrile / water (1:1) and diluted with methanol / 12.5 mM phosphate buffer pH 3.4 (4:1), then filtered (0.2 μm RC, Lab Logistics Group, Germany) and further analyzed by UPLC (n=2).

[0285] The UPLC assay was set up according to the following

[0286] Table 1.

[0287] Table 1. UPLC method parameters.

[0288]

[0289] The concentration of the injected sample was automatically calculated by the software provided by the manufacturer. The drug loading was calculated according to the following formula, where C 来那度胺 is the analyzed concentration of lenalidomide, m 样品 For the sample mass:

[0290]

[0291] result

[0292] The lenalidomide drug loading of the particles of Comparative Example 1 was determined to be 94.8%.

[0293] The lenalidomide drug loading of the particles of Comparative Example 2 was determined to be 92.6%.

[0294] The lenalidomide drug loading of the particles of Example 3 was determined to be 76.0%.

[0295] The lenalidomide drug loading of the particles of Example 4 was determined to be 67.7%.

[0296] Example 6

[0297] Coating integrity

[0298] This example describes the analysis of the coating integrity of the granules obtained in Comparative Examples 1 and 2 and Examples 3 and 4.

[0299] Coating integrity of coated lenalidomide was determined by preparing a suspension of the coating material in DMSO, a solvent that dissolves the API but not the coating material. Therefore, release of lenalidomide from the product can only occur due to a "defect" in the coating. The integrity of the coating can be assessed by using an HPLC method to measure the released lenalidomide. The lower the percentage of released lenalidomide, the better the integrity of the coating.

[0300] Materials and methods

[0301] The coating integrity (i.e., w / w % of lenalidomide released from the powder) was determined using UPLC (using a Prominence-i (Shimadzu, Japan) equipped with a diode array detector (Shimadzu, Japan) set at a wavelength of 223 nm, using a 4.6×100 mm, 2.6 μm particle, C18WP column (SunShell, ChromaNik Technologies Inc, Osaka, Japan). 25 mg of the material was dispersed in 25 mL of DMSO and placed on a turntable for 3 h. A 1 mL sample was removed for filtration (0.2 μm RC, Lab Logistics Group, Germany) and further analyzed using UPLC (n=1) according to the settings in Table 1 above.

[0302] The concentration of the injected sample was automatically calculated by the software provided by the manufacturer. The amount of API dissolved in the coating integrity assay was calculated according to the following formula, where c a is the API concentration (mg / ml) obtained by analysis, V t is the total sample volume (ml), m API is the weighed API mass in (mg), and drug loading is the API content (fraction) in the coated API:

[0303]

[0304] result

[0305] The amount of dissolved API in the coating integrity assay of the granules according to Comparative Example 1 was determined to be 74%.

[0306] The amount of dissolved API in the coating integrity assay of the granules according to Comparative Example 2 was determined to be 51%.

[0307] The amount of dissolved API in the coating integrity assay of the granules according to Example 3 was determined to be 15%.

[0308] The amount of dissolved API in the coating integrity assay of the granules according to Example 4 was determined to be 7%.

[0309] Example 7

[0310] Preparation of formulations

[0311] This example describes the preparation of a formulation of coated lenalidomide particles as disclosed herein.

[0312] Materials and methods

[0313] Suspensions of coated microparticles of lenalidomide according to Comparative Examples 1 and 2 and Examples 3 and 4 were prepared.

[0314] The powder of coated micronized particles of lenalidomide according to Comparative Examples 1 and 2 and Examples 3 and 4 was mixed with vet (Boehringer Ingelheim Animal Health, France) were mixed together. Vet is a veterinary medical product for animal injection, which contains 10 mg / mL of sodium hyaluronate in a sterile, isotonic phosphate buffer solution (pH 7.4). The composition of vet is presented in Table 2 below.

[0315] Table 2: Composition of vehicles used to suspend particles as disclosed herein.

[0316]

[0317]

[0318] Reconstitute the microparticles by adding vehicle to the vial of microparticle powder to obtain the following lenalidomide concentrations:

[0319] - 50 mg / ml lenalidomide according to the particles of Comparative Example 1,

[0320] - 5 mg / ml and 50 mg / ml lenalidomide according to the granules of Comparative Example 2,

[0321] - 20 mg / ml lenalidomide according to the particles of Example 3, and

[0322] - 20 mg / ml lenalidomide according to the particles of Example 4.

[0323] result

[0324] Suspensions of coated lenalidomide particles according to Comparative Examples 1 and 2 and Examples 3 and 4, respectively, were obtained.

[0325] Comparative Example 8

[0326] In vivo pharmacokinetic studies I

[0327] This example describes a preclinical pharmacokinetic study of lenalidomide following administration of coated lenalidomide formulations according to Comparative Examples 1 and 2 in male and female Sprague Dawley rats.

[0328] Materials and methods

[0329] Male and female Sprague Dawley rats weighing between 231 and 285 g on the day of administration were provided by Charles River (UK). The study duration was 14 days. 36 rats were used in the study.

[0330] The hair of the application area was clipped before injection, and the injection site was marked. The suspension prepared with the coated lenalidomide particles according to Comparative Examples 1 and 2 as described in Example 7 was drawn into a 1 mL BD syringe and a single subcutaneous injection (about 0.12 mL) was administered into the flank of each rat through a 23G needle (BD Microlance).

[0331] Blood samples (about 0.2 mL) were collected from the tail vein into K2EDTA tubes at the following time points: 0.25, 0.5, 1, 2, 3, 6, 12, 24, 48, 72, 120, 168, 264, and 336 h after administration. Plasma was separated as soon as possible by centrifugation (1500 g, 4 ° C, for 10 min). Animals were killed on the last day of the study. Plasma samples were stored in a refrigerator at -80 ° C awaiting analysis.

[0332] Plasma concentrations were determined using UPLC-MS / MS.

[0333] Samples were prepared by pipetting 35 μL rat plasma into 96-well plates using a TECAN Genesis liquid handling robot, adding 35 μL 5% DMF solution in acetonitrile, 70 μL internal standard working solution. The 96-well plates were shaken for 15 minutes and centrifuged. All samples were then injected into a UPLC-MS / MS system (Xevo TQ-s micro was connected to an Acquity I-Class UPLC system, Waters, Milford, MA, USA), wherein the settings were as shown in Table 3 below.

[0334] Table 3. Parameters of the UPLC-MS / MS method.

[0335] column Acquity CSH C18(50x2.1mm,1.7μm) Mobile phase A Water with 0.10% formic acid Mobile phase B ACN:DMSO(95:5) Washing solution ACN:MeOH (1:1)

[0336] Pharmacokinetic analysis of lenalidomide in plasma was performed using PKanalix (2021, Lixoft, Antony, France) according to standard non-compartmental methods. Dose-normalized plasma concentrations of lenalidomide were assessed after a single subcutaneous administration of the formulation, and the following plasma pharmacokinetic parameters were evaluated:

[0337] • Doses will be expressed in mg / kg rat body weight.

[0338] ·“C max ”: Maximum concentration found in the analysis, expressed in ng / mL.

[0339] · “AUC ∞ ”: Area under the concentration versus time curve up to infinite time, expressed in ng*h / mL.

[0340] ·“C max / D”: maximum concentration normalized to 1 mg / kg, expressed as ng / mL / mg / kg rat body weight.

[0341] · “AUC ∞ / D”: Area under the concentration versus time curve normalized to 1 mg / kg until infinity, expressed as ng*h / mL / mg / kg rat body weight.

[0342] · “Fr.Rel. 0-12h ”: The fraction released during the first twelve hours of the area under the concentration versus time curve up to infinity, expressed as a percentage.

[0343] result

[0344] Figure 1 and Figure 2 The corresponding plasma concentration-time curves (lenalidomide plasma concentration (ng / mL) versus sampling time (h)) of the samples obtained by Comparative Examples 1 and 2, respectively, are shown.

[0345] Table 4. Results of evaluation of plasma pharmacokinetic parameters.

[0346]

[0347] Example 9

[0348] In vivo pharmacokinetic studies II

[0349] This example describes a preclinical pharmacokinetic study of lenalidomide following administration of coated lenalidomide formulations according to Examples 3 and 4 in male and female Sprague Dawley rats.

[0350] Materials and methods

[0351] Male Sprague Dawley rats weighing between 260 and 343 g on the day of administration were provided by Charles River (UK). The study duration was 14 days. 44 rats were used in the study.

[0352] The hair of the application area was clipped before injection, and the injection site was marked. The suspension prepared with the coated lenalidomide particles according to Comparative Examples 3 and 4 as described in Example 7 was drawn into a 1 mL BD syringe and a single subcutaneous injection (about 0.15 mL) was administered into the flank of each rat through a 23G needle (BD Microlance).

[0353] Blood samples (about 0.2 mL) were collected from the tail vein into K2EDTA tubes at the following time points: 0.25, 0.5, 1, 2, 3, 6, 12, 24, 48, 72, 120, 168, 216, 264, and 336 h after administration. Plasma was separated as soon as possible by centrifugation (1500 g, 4 ° C, for 10 min). Animals were killed on the last day of the study. Plasma samples were stored in a refrigerator at -80 ° C awaiting analysis.

[0354] Plasma concentrations were determined using UPLC-MS / MS.

[0355] Samples were prepared by pipetting 35 μL rat plasma into a 96-well plate using a TECAN Genesis liquid handling robot, adding 35 μL 5% DMF solution in acetonitrile, 70 μL internal standard working solution. The 96-well plate was shaken for 15 minutes and centrifuged. All samples were then injected into a UPLC-MS / MS system (Xevo TQ-s micro coupled to an Acquity I-Class UPLC system, Waters, Milford, MA, USA), with settings as shown in Table 2 above.

[0356] Pharmacokinetic analysis of lenalidomide in plasma was performed according to standard non-compartmental methods using Phoenix WinNonlin version 8.3 (Certara, USA). Dose-normalized plasma concentrations of lenalidomide were assessed after a single subcutaneous administration of the formulation, and the following plasma pharmacokinetic parameters were evaluated:

[0357] • Doses will be expressed in mg / kg rat body weight.

[0358] ·“C max ”: Maximum concentration found in the analysis, expressed in ng / mL.

[0359] · “AUC ∞ ”: Area under the concentration versus time curve up to infinite time, expressed in ng*h / mL.

[0360] ·“C max / D”: maximum concentration normalized to 1 mg / kg, expressed as ng / mL / mg / kg rat body weight.

[0361] · “AUC∞ / D”: Area under the concentration versus time curve normalized to 1 mg / kg until infinity, expressed as ng*h / mL / mg / kg rat body weight.

[0362] · “Fr.Rel. 0-12h ”: The fraction released during the first twelve hours of the area under the concentration versus time curve up to infinity, expressed as a percentage.

[0363] result

[0364] Figure 3 and Figure 4 The corresponding plasma concentration-time curves (lenalidomide plasma concentration (ng / mL) versus sampling time (h)) of the samples obtained by Examples 3 and 4, respectively, are shown.

[0365] Table 5. Results of evaluation of plasma pharmacokinetic parameters.

[0366]

[0367]

[0368] The dose normalized maximum concentration of Example 3 was lower compared to Comparative Example 1, as indicated by both the lower maximum concentration and the lower fraction released during the first twelve hours.

[0369] The dose normalized maximum concentration of Example 4 was lower compared to Comparative Example 2, as indicated by both the lower maximum concentration and the lower fraction released during the first twelve hours.

[0370] In summary, Comparative Example 8 and Example 9 demonstrate that coated lenalidomide particles produced by stopped-flow ALD show a release profile with lower initial release than coated lenalidomide particles produced using continuous-flow ALD.

[0371] Comparative Example 10

[0372] Coating of Indomethacin Microparticles Using Continuous Flow ALD I

[0373] Indomethacin microparticles 99.9% (ReechPharma, CA, USA) were used as received. The average diameter of the indomethacin particles was 10.5 μm as determined by laser diffraction (Shimadzu, SALD-7500 nano, Kyoto, Japan). The particle size distribution determined by laser diffraction was as follows: 10 4.2μm; D 50 10.5 μm and D 90 23.8μm.

[0374] Microparticles or solid cores are coated as described in steps 1 to 4.

[0375] 1. Load the micron particles into the ALD reactor (Picosun, SUNALE TM R series, Espoo, Finland), and three ALD cycles using diethylzinc and water as precursors were performed at a reactor temperature of 50°C, followed by one ALD cycle with trimethylaluminum and water as precursors. This was repeated six times, i.e., 6x (3 Zn cycles + 1 Al cycle), resulting in a total of 24 cycles. By this method, a first layer of a mixed oxide having a zinc:aluminum atomic ratio of approximately 3:1 was formed.

[0376] The ALD reactor includes a reaction chamber into which the micron particles are loaded. The ALD reactor further includes precursor bottles that independently contain each precursor, each precursor bottle being connected to the reaction chamber via a valve. The ALD reactor also includes a pump and associated piping for pumping an inert gas, such as nitrogen, through the reaction chamber, the pump also being connected to the reaction chamber via a valve.

[0377] The ALD cycle is performed as follows, where steps a to e represent the first cycle, subsequent cycles begin with step d as specified in step 1, and the final cycle ends at step k:

[0378] a. Reagent pulse: Open the valve of the precursor bottle for 0.1s, evaporate the water and bring it into the reaction chamber by inert nitrogen gas. The water is adsorbed onto the surface of the drug particles, presenting hydroxyl groups on the outside of the particles.

[0379] b. The reactor was then pumped for 3 s.

[0380] c. Repeat steps a to b above 100 times.

[0381] d. Purge pulse: Purge the chamber with nitrogen. Gaseous water and organic gases (in case this is not the first cycle) will be removed.

[0382] e. Reagent pulse: The valve of the precursor bottle is opened for 0.1 s, diethylzinc and trimethylaluminum are evaporated and carried into the reaction chamber by inert nitrogen. Dimethylzinc or trimethylaluminum is adsorbed onto the surface of the drug particles and reacts with the hydroxyl groups. This releases ethane from diethylzinc or methane from trimethylaluminum.

[0383] f. The reactor was then pumped for 3 s.

[0384] g. Repeat steps e to f above 100 times.

[0385] h. Purge pulse: Purge the chamber with nitrogen to remove unreacted reagents and organic gases.

[0386] i. Reagent pulse: The valve of the precursor bottle is opened for 0.1 s, and water is evaporated and carried into the reaction chamber by inert nitrogen. Water is adsorbed to the surface of the drug particles and reacts with the metal organic surface. The remaining ethyl or methyl groups are converted to ethane and methane, respectively. As a result, the surface is covered with a metal oxide layer, presenting hydroxyl groups on the outside of the particles.

[0387] j. After this the reactor was pumped for 3 s.

[0388] k. Repeat the above steps i to j 100 times

[0389] l. Repeat the cycle starting from step d.

[0390] 2. Next, the powder was taken out from the reactor and deagglomerated by a sonic sieving machine (Tsutsui sonic stirring sieving machine SW-20AT) with a 20 μm mesh size sieve.

[0391] 3. The resulting deagglomerated powder was reloaded into the ALD reactor and steps 1 to 2 were repeated twice to form a second and third layer of a mixed oxide having an approximately 3:1 zinc:aluminum atomic ratio.

[0392] 4. The resulting deagglomerated powder was reloaded into the ALD reactor and step 1 was repeated once.

[0393] Example 11

[0394] Coating of Indomethacin Microparticles Using Stopped Flow I

[0395] Indomethacin microparticles 99.9% (ReechPharma, CA, USA) were used as received. The average diameter and particle size distribution were as described for Comparative Example 10.

[0396] The microparticles were coated essentially as described in steps 1 to 4 of Comparative Example 10, except that the ALD cycles were performed as follows (steps a to d represent the first cycle, with subsequent cycles starting from step a as specified in step e).

[0397] a. Reagent pulse 1:

[0398] i. Close the valve on the pipe between the pump and the ALD reactor.

[0399] ii. Open the valve on the water precursor bottle for 1 s to allow the evaporated water to fill the reaction chamber.

[0400] iii. Close the valve of the water precursor bottle and let the chamber sit for 15 s (soak time) before opening the pump valve again to ensure that the water vapor adsorbs to the surface of the drug particles, presenting hydroxyl groups on the outside or particles.

[0401] iv. The reactor was then pumped for 9 s.

[0402] v. Repeat steps i to iv above 20 times.

[0403] b. Purge pulse: Purge the chamber with nitrogen in a continuous flow. Gaseous water and organic gases (in case this is not the first cycle) will be removed.

[0404] c. Reagent pulse:

[0405] i. Close the valve on the pipe between the pump and the ALD reactor.

[0406] ii. The valve on the diethylzinc or trimethylaluminum precursor bottle was opened for 1 second to allow the evaporated metal-containing precursor to fill the reaction chamber.

[0407] iii. The valve of the precursor bottle was closed and the chamber was left to stand for 15 s (soak time) before opening it to the pump again to ensure that the metal-containing precursor vapor reacted with the hydroxyl groups on the surface of the drug particles.

[0408] iv. The reactor was then pumped for 9 s.

[0409] v. Repeat steps i-iv above 20 times.

[0410] d. Purge pulse: Purge the chamber with nitrogen in a continuous flow to remove unreacted reagents and organic gases.

[0411] e. Repeat the cycle starting from step a.

[0412] Example 12

[0413] Determination of drug loading

[0414] This example describes the analysis of the indomethacin drug loading of the particles obtained in Example 11.

[0415] Materials and methods

[0416] To determine the drug loading (i.e., w / w % of indomethacin in the powder), UPLC (Nexera, Shimadzu, Japan) equipped with a diode array detector (Shimadzu, Japan) set at a wavelength of 254 nm was used using a 3×100 mm, 2.6 μm particle, phenylhexyl column (Thermo Fisher Scientific Inc., MA, USA). The material was dissolved in 2 M phosphoric acid in acetonitrile / water (1:1) and diluted with acetonitrile / water (3:1), then filtered (0.2 μm RC, Lab Logistics Group, Germany) and further analyzed using UPLC (n=3).

[0417] The UPLC assay was set up according to Table 6.

[0418] Table 6. UPLC method parameters.

[0419]

[0420]

[0421] The concentration of the injected sample was automatically calculated by the software provided by the manufacturer. The drug loading was calculated according to the following formula, where C 样品 is the analytical concentration of indomethacin, V t is the total volume of the sample, m 样品 For the sample mass:

[0422] Drug loading (%) = 100·(C 样品 ·V t ) / m 样品

[0423] result

[0424] The indomethacin drug loading of the granules according to Example 11 was determined to be 83.2%.

[0425] Example 13

[0426] Determination of coating integrity

[0427] This example describes the analysis of the coating integrity of the granules obtained in Example 11.

[0428] The coating integrity of coated indomethacin was determined by preparing a suspension of the coating material in DMSO, a solvent that dissolves the API but not the coating material. Therefore, the release of indomethacin from the product can only occur due to a "defect" in the coating. The integrity of the coating can be assessed by using an HPLC method to measure the released indomethacin. The lower the percentage of released indomethacin, the better the integrity of the coating.

[0429] Materials and methods

[0430] To determine coating integrity (i.e., w / w% indomethacin in powder), a UPLC (Nexera, Shimadzu, Japan) equipped with a diode array detector (Shimadzu, Japan) set at a wavelength of 254 nm was used using a 3×100 mm, 2.6 μm particle, phenylhexyl column (Thermo Fisher Scientific Inc., MA, USA). 25 mg of the material was dispersed in 25 mL of DMSO and placed on a turntable for 3 h. A 1 mL sample was removed for filtration (0.2 μm RC, Lab Logistics Group, Germany) and further analyzed using UPLC (n=1) according to the settings in Table 6 above.

[0431] The concentration of the injected sample was automatically calculated by the software provided by the manufacturer. The amount of API dissolved in the coating integrity assay was calculated according to the following formula, where c a is the API concentration (mg / ml) obtained by analysis, V t is the total sample volume (ml), m API is the weighed API mass in (mg), and drug loading is the API content (fraction) in the coated API:

[0432] API dissolved amount (%) = 100·(c a ·V t ) / (m API Drug loading)

[0433] result

[0434] The amount of dissolved API in the coating integrity assay of the granules according to Example 11 was determined to be 13.0%.

[0435] Example 14

[0436] Coating of Indomethacin Microparticles Using Stop-Flow II

[0437] Indomethacin microparticles 99.9% (ReechPharma, CA, USA) were used as received. The average diameter and particle size distribution were as described for Comparative Example 10.

[0438] The microparticles were coated essentially as described in steps 1 to 4 of Example 11, except that the immersion time in steps a and c was 30 s instead of 15 s.

[0439] Example 15

[0440] Determination of drug loading

[0441] This example describes the analysis of the indomethacin drug loading of the particles obtained in Example 14.

[0442] Materials and methods

[0443] Materials and Methods were as described in Example 12, except that the particles analyzed were those obtained in Example 14 rather than those obtained in Example 11.

[0444] result

[0445] The indomethacin drug loading of the granules according to Example 14 was determined to be 82.3%.

[0446] Example 16

[0447] Determination of coating integrity

[0448] This example describes the analysis of the coating integrity of the granules obtained in Example 14.

[0449] The coating integrity of the coated indomethacin was determined as described in Example 13.

[0450] Materials and methods

[0451] Materials and Methods were as described in Example 13, except that the particles analyzed were those obtained in Example 14 rather than those obtained in Example 11.

[0452] result

[0453] The amount of dissolved API in the coating integrity assay of the granules according to Example 14 was determined to be 10.6%.

[0454] Comparative Example 17

[0455] Coating of Lenalidomide Microparticles Using Continuous Flow ALD

[0456] This is similar to Comparative Example 10, except that Lenalidomide Microparticles 99.9% (Kyongbo, South Korea) were used as received instead of Indomethacin 99.9%. The average diameter of the Lenalidomide particles was 4.3 μm, as determined by laser diffraction (Shimadzu, SALD-7500nano, Kyoto, Japan). The particle size distribution determined by laser diffraction was as follows: D10 0.9 μm; D50 4.3 μm and D90 15.6 μm.

[0457] All other aspects of the method used in Comparative Example 17 were the same as those of Comparative Example 10 except for the microparticles used.

[0458] Example 18

[0459] Coating of Lenalidomide Microparticles Using Stopped Flow I

[0460] Lenalidomide microparticles 99.9% (Kyongbo, South Korea) were used as received. The average diameter of the lenalidomide particles was 4.3 μm, as described above in Comparative Example 17. The particle size distribution determined by laser diffraction was as follows: D10 0.9 μm; D50 4.3 μm and D90 15.6 μm.

[0461] The microparticles were coated essentially as described in steps 1 to 4 of Comparative Example 17, except that:

[0462] - in step 3, steps 1 to 2 are repeated four times to form a second layer, a third layer, a fourth layer and a fifth layer of a mixed oxide having a zinc:aluminum atomic ratio of about 3:1; and

[0463] - The ALD cycle is performed as follows (steps a to d represent the first cycle, subsequent cycles start from step a as specified in step e).

[0464] a. Reagent pulse 1:

[0465] i. Close the valve on the pipe between the pump and the ALD reactor.

[0466] ii. Open the valve on the water precursor bottle for 1 s to allow the evaporated water to fill the reaction chamber.

[0467] iii. Close the valve of the water precursor bottle and let the chamber stand for 30 s (soak time) before opening the pump valve again to ensure that the water vapor adsorbs to the surface of the drug particles, presenting hydroxyl groups on the outside or particles.

[0468] iv. The reactor was then pumped for 9 s.

[0469] v. Repeat steps i to iv above 20 times.

[0470] b. Purge pulse: Purge the chamber with nitrogen in a continuous flow. Gaseous water and organic gases (in case this is not the first cycle) will be removed.

[0471] c. Reagent pulse:

[0472] i. Close the valve on the pipe between the pump and the ALD reactor.

[0473] ii. The valve on the diethylzinc or trimethylaluminum precursor bottle was opened for 1 second to allow the evaporated metal-containing precursor to fill the reaction chamber.

[0474] iii. The valve of the precursor bottle was closed and the chamber was left to stand for 30 s (soak time) before opening it to the pump again to ensure that the metal-containing precursor vapor reacted with the hydroxyl groups on the surface of the drug particles.

[0475] iv. The reactor was then pumped for 9 s.

[0476] v. Repeat steps i-iv above 20 times.

[0477] d. Purge pulse: Purge the chamber with nitrogen in a continuous flow to remove unreacted reagents and organic gases.

[0478] e. Repeat the cycle starting from step a.

[0479] Example 19

[0480] Determination of drug loading

[0481] This example describes the analysis of the lenalidomide drug loading of the particles obtained in Example 18.

[0482] Materials and methods

[0483] Materials and Methods were as described in Example 12, except that the particles analyzed were those obtained in Example 18 rather than those obtained in Example 11.

[0484] result

[0485] The lenalidomide drug loading of the particles of Example 18 was determined to be 69.2%.

[0486] Example 20

[0487] Determination of coating integrity

[0488] This example describes the analysis of the coating integrity of the granules obtained in Example 18.

[0489] The coating integrity of the coated lenalidomide was determined as described in Example 13.

[0490] Materials and methods

[0491] Materials and Methods were as described in Example 13, except that the particles analyzed were those obtained in Example 18 rather than those obtained in Example 11.

[0492] result

[0493] The amount of dissolved API in the coating integrity assay of the granules according to Example 18 was determined to be 6.8%.

[0494] Example 21

[0495] Preparation of formulations

[0496] This example describes the preparation of a formulation of coated lenalidomide particles as disclosed herein.

[0497] The powder of coated microparticles of lenalidomide according to the example was mixed with vet (Boehringer Ingelheim Animal Health, France) were mixed together. Vet is a veterinary medical product for animal injection, which contains 10 mg / mL of sodium hyaluronate in a sterile, isotonic phosphate buffer solution (pH 7.4). The composition of vet is presented in Table 9 below.

[0498] Table 9: Composition of vehicles used to suspend particles as disclosed herein.

[0499] Sodium Hyaluronate 10mg Active substances Sodium chloride 8.5mg Isotonicity Agents Sodium Acid Phosphate 0.223mg Buffer Sodium dihydrogen phosphate monohydrate 40 μg Buffer hydrochloric acid qs For pH adjustment Sodium hydroxide qs For pH adjustment Water for injection qsad 1mL Solvents

[0500] Reconstitute the microparticles by adding vehicle to the vial of microparticle powder to obtain the following lenalidomide concentrations:

[0501] - 20 mg / mL lenalidomide according to the particles of Example 18.

[0502] Example 22

[0503] In vivo pharmacokinetic studies

[0504] This example describes a preclinical pharmacokinetic study of lenalidomide following administration of a coated lenalidomide formulation according to Example 21 in male and female Sprague Dawley rats.

[0505] Materials and methods

[0506] Male and female Sprague Dawley rats weighing between 260 and 343 g on the day of administration were provided by Charles River (UK). The study duration was 14 days. Three rats of each sex were used in each study group of the study. The study groups were the coated microparticle formulation according to Example 21, and uncoated microparticles of lenalidomide as a control.

[0507] The hair of the application area was clipped before injection and the injection site was marked. The suspension prepared with the coated lenalidomide particles according to Example 18 as described in Example 21 was drawn into a 1 mL BD syringe and a single subcutaneous injection (about 0.12 mL) was administered into the flank of each rat through a 23G needle (BD Microlance).

[0508] Blood samples (about 0.2 mL) were collected from the tail vein into K2EDTA tubes at the following time points: 0.25, 0.5, 1, 2, 3, 6, 12, 24, 48, 72, 120, 168, 264, and 336 h after administration. Plasma was separated as soon as possible by centrifugation (1500 g, 4 ° C, for 10 min). Animals were killed on the last day of the study. Plasma samples were stored in a refrigerator at -80 ° C awaiting analysis.

[0509] Plasma concentrations were determined using UPLC-MS / MS.

[0510] Using TECAN EVO Ware liquid handling robot, 35 μL rat plasma was pipetted into 384-well plates, and 35 μL 5% DMSO solution in acetonitrile and 70 μL internal standard working solution were added to prepare samples. The 384-well plates were shaken for 15 minutes and centrifuged. All samples were then injected into a UPLC-MS / MS system (Xevo TQ-s micro connected to an Acquity I-Class UPLC system, Waters, Milford, MA, USA), which was set as shown in Table 10 below.

[0511] Table 10. Parameters of the UPLC-MS / MS method.

[0512] column Acquity CSH C18(50x2.1mm,1.7μm) Mobile phase A Water with 0.10% formic acid Mobile phase B ACN:DMSO(95:5) Washing solution ACN:MeOH (1:1)

[0513] The pharmacokinetic analysis of lenalidomide in plasma was performed using the software Phoenix WinNonlin version 8.3 (Certara, USA) based on the non-compartmental analysis (NCA) used. The dose-normalized plasma concentrations of lenalidomide after a single subcutaneous administration of the formulation were assessed, and the following plasma pharmacokinetic parameters were evaluated:

[0514] ·C max -Maximum observed plasma concentration (ng / mL)

[0515] ·T max -Achieve C max Time (h)

[0516] AUC 0-24h - AUC (h.ng / mL) from time 0 to time 24 hours.

[0517] AUC last - AUC (h.ng / mL) from time 0 to the time of the last detectable or last measured (336h) plasma concentration.

[0518] Nominal plasma sampling time points and nominal doses were used for noncompartmental PK analysis. PK of the animals was calculated using the extravascular dose option in WinNonlin. Reported plasma concentrations below the LLOQ but above 3 ng / mL were included in the PK analysis, while unreported plasma concentrations below the LLOQ were omitted from the analysis.

[0519] C max and T max Derived from observed plasma concentration data. AUC was estimated by integrating the plasma concentration versus time curve and using linear interpolation for increasing plasma levels and logarithmic interpolation for decreasing plasma levels (Linear Up Log Down method).

[0520] result

[0521] Figure 5 The plasma concentration-time curves (lenalidomide plasma concentration (ng / mL) versus sampling time (h)) of the sample obtained by Example 21 and the sample obtained using uncoated lenalidomide microparticles as a control are shown.

[0522] More specifically, the change in plasma concentration (ng / mL) of lenalidomide over time (h) is shown in a semi-logarithmic graph (y is a logarithmic scale). The squares show the results from female rats (n=3), and the circles show the results from male rats (n=3). The open symbols show the results from the control (uncoated lenalidomide microparticles), and the solid symbols show the results from the coated lenalidomide microparticles according to Example 21.

[0523] Table 11. Results of evaluation of plasma pharmacokinetic parameters, mean values ​​for both male and female rats (n=6).

[0524] parameter Comparison Example 21 <![CDATA[T max ]]> 0,5 0,8 <![CDATA[C max ]]> 1610 152 <![CDATA[AUC 0-24h ]]> 5310 520 <![CDATA[AUC last ]]> 5520 4330

[0525] The maximum concentration of Example 21 was lower and reached more slowly than the control.

[0526] AUC 0-24h and AUC lastRepresent the amount of lenalidomide released after 24 hours and 336 hours, respectively. For the control, more than 96% of the total amount of lenalidomide released was released within the first 24 hours. In contrast, for Example 21, the amount of lenalidomide released after 24 hours was approximately 12% of the amount released after 336 hours, and as Figure 5 As shown, drug release is not yet complete.

[0527] Accordingly, Example 21 demonstrates that coated lenalidomide particles produced by stopped-flow ALD exhibit a release profile with a lower initial release than uncoated lenalidomide particles.

[0528] Example 23

[0529] Coating of Lenalidomide Microparticles Using Stop-Flow II

[0530] Lenalidomide micronized particles 99.9% (Kyongbo, South Korea) were used as received. The average diameter of the lenalidomide particles was 4 μm, as described in Comparative Example 17 above. The particle size distribution determined by laser diffraction was as follows: 10 0.6μm; D 50 3.9μm and D 90 15.6μm.

[0531] The microparticles were coated essentially as described in Example 18, except that the layers were formed in alternating 4:1 (Zn:Al) and 3:1 (Zn:Al) groups rather than a uniform 3:1 (ZN:Al) group. Accordingly, Groups 1, 3, and 5 each included a total of 25 cycles, with a ratio of 4 Zn cycles: 1 Al cycle. Meanwhile, Groups 2, 4, and 6 each included a total of 24 cycles, with a ratio of 3 Zn cycles: 1 Al cycle.

[0532] Example 24

[0533] Determination of drug loading

[0534] This example describes the analysis of the lenalidomide drug loading of the particles obtained in Example 23.

[0535] Materials and methods

[0536] Materials and Methods were as described in Example 12, except that the particles analyzed were those obtained in Example 23 rather than those obtained in Example 11.

[0537] result

[0538] The lenalidomide drug loading of the particles of Example 23 was determined to be 75.2%.

[0539] Example 25

[0540] Determination of coating integrity

[0541] This example describes the analysis of the coating integrity of the granules obtained in Example 23.

[0542] The coating integrity of the coated lenalidomide was determined as described in Example 13.

[0543] Materials and methods

[0544] Materials and Methods were as described in Example 13, except that the particles analyzed were those obtained in Example 23 rather than those obtained in Example 11.

[0545] result

[0546] The amount of dissolved API in the coating integrity assay of the granules according to the example was determined to be 16.2%.

[0547] Example 26

[0548] Preparation of formulations

[0549] For this example, a formulation of coated lenalidomide particles disclosed herein was prepared as described in Example 21, except that the particles obtained in Example 23 were used instead of the particles obtained in Example 18.

[0550] Example 27

[0551] In vivo pharmacokinetic studies

[0552] This example describes a preclinical pharmacokinetic study of lenalidomide following administration of a coated lenalidomide formulation according to Example 26 in male and female Sprague Dawley rats.

[0553] Materials and methods

[0554] Male and female Sprague Dawley rats weighing between 260 and 343 g on the day of administration were provided by Charles River (UK). The study duration was 14 days. In each study group of the study, 3 rats of each sex were used. The study groups were the coated microparticle formulations according to the examples, and uncoated microparticles of lenalidomide as a control.

[0555] The hair of the application area was clipped before injection and the injection site was marked. The suspension prepared with the coated lenalidomide particles according to Example 23 as described in Example 26 was drawn into a 1 mL BD syringe and a single subcutaneous injection (about 0.12 mL) was administered into the flank of each rat through a 23G needle (BD Microlance).

[0556] Blood samples (about 0.2 mL) were collected from the tail vein into K2EDTA tubes at the following time points: 0.25, 0.5, 1, 2, 3, 6, 12, 24, 48, 72, 120, 168, 264, and 336 h after administration. Plasma was separated as soon as possible by centrifugation (1500 g, 4 ° C, for 10 min). Animals were killed on the last day of the study. Plasma samples were stored in a refrigerator at -80 ° C awaiting analysis.

[0557] Plasma concentrations were determined using UPLC-MS / MS.

[0558] Using TECAN EVO Ware liquid handling robot, 35 μL rat plasma was pipetted into 384-well plates, 35 μL 5% DMSO solution in acetonitrile, 70 μL internal standard working solution were added to prepare samples. The 384-well plates were shaken for 15 minutes and centrifuged. All samples were then injected into a UPLC-MS / MS system (Xevo TQ-s micro connected to an Acquity I-Class UPLC system, Waters, Milford, MA, USA), which was set as shown in Table 12 below.

[0559] Table 12. Parameters of the UPLC-MS / MS method.

[0560] column Acquity CSH C18(50x2.1mm,1.7μm) Mobile phase A Water with 0.10% formic acid Mobile phase B ACN:DMSO(95:5) Washing solution ACN:MeOH (1:1)

[0561] The pharmacokinetic analysis of lenalidomide in plasma was performed using the software Phoenix WinNonlin version 8.3 (Certara, USA) based on the non-compartmental analysis (NCA) used. The dose-normalized plasma concentrations of lenalidomide after a single subcutaneous administration of the formulation were assessed, and the following plasma pharmacokinetic parameters were evaluated:

[0562] ·C max -Maximum observed plasma concentration (ng / mL)

[0563] ·T max -Achieve C max Time (h)

[0564] AUC 0-24h - AUC (h.ng / mL) from time 0 to time 24 hours.

[0565] AUC last - AUC (h.ng / mL) from time 0 to the time of the last detectable or last measured (336h) plasma concentration.

[0566] Nominal plasma sampling time points and nominal doses were used for noncompartmental PK analysis. PK of the animals was calculated using the extravascular dose option in WinNonlin. Reported plasma concentrations below the LLOQ but above 3 ng / mL were included in the PK analysis, while unreported plasma concentrations below the LLOQ were omitted from the analysis.

[0567] C max and T max Derived from observed plasma concentration data. AUC was estimated by integrating the plasma concentration versus time curve and using linear interpolation for increasing plasma levels and logarithmic interpolation for decreasing plasma levels (ascending linear-declining logarithmic method).

[0568] result

[0569] Figure 6 The plasma concentration-time curves (lenalidomide plasma concentration (ng / mL) versus sampling time (h)) of the sample obtained by Example 26 and the sample obtained using uncoated lenalidomide microparticles as a control are shown.

[0570] More specifically, the change in plasma concentration (ng / mL) of lenalidomide over time (h) is shown in a semi-logarithmic graph (y is a logarithmic scale). The squares show the results from female rats (n=3), and the circles show the results from male rats (n=3). The open symbols show the results from the control (uncoated lenalidomide microparticles), and the solid symbols show the results from the coated lenalidomide microparticles according to Example 26.

[0571] Table 13. Plasma pharmacokinetic parameter evaluation results are averaged for male and female rats (n=6).

[0572] parameter Comparison Example 26 <![CDATA[T max ]]> 0,5 0,5 <![CDATA[C max ]]> 1610 392 <![CDATA[AUC 0-24h ]]> 5310 1217 <![CDATA[AUC last ]]> 5520 4892

[0573] The maximum concentration of Example 21 was lower compared to the control.

[0574] For the control, more than 96% of the total amount of lenalidomide released was released within the first 24 hours. In contrast, for Example 26, the amount of lenalidomide released after 24 hours was approximately 25% of the amount released after 336 hours, and as Figure 6 As shown, drug release is not yet complete.

[0575] Accordingly, Example 26 demonstrates that coated lenalidomide particles produced by stopped-flow ALD exhibit a release profile with a lower initial release than uncoated lenalidomide particles.

[0576] Example 28

[0577] Stop-flow coated liraglutide microparticles

[0578] R&D grade liraglutide (MedChem Express, New Jersey, US) with a purity of 98.5% and a peptide content of 90.8% was suspended in a 0.1% Span 85 (Sigma-Aldrich, MO, USA) solution in cyclohexane (Merck, Germany). The particle size distribution determined by laser diffraction (SALD-7500 nano (Shimadzu, Japan), 405 nm laser) was as follows: %D (10): 2.0 μm, %D (50): 7.3 μm, %D (90): 23.8 μm.

[0579] The raw materials were dried in a small spray dryer (B-290, dehumidifier B-296 with two-fluid nozzle; The raw materials were dispersed in purified water (0.8-2MΩ / cm2) to form a milky white liquid with a liraglutide concentration of 15wt%. The dispersion was spray dried at an inlet temperature of 115°C and a suction rate of 100% (~35m 3 / h), pump rate 8% (3.8 mL / min), nozzle cleaning 2, volume flow 35 mm (N2, ~600 L / h), outlet temperature 73 ° C. The total mass yield was about 77.5%. The spray-dried material was analyzed using a Nexera UPLC-UV-DAD (Shimadzu, Japan) with a SunShell nC18-WP, 4.6×100 mm, 2.6 μm particle size column (Chromanik Technologies Inc., Japan) and was found to have a liraglutide content of 100.1±0.2% (relative) in the spray-dried material. The particle size of the spray-dried material determined as described above was as follows: %D(10): 1.7 μm, %D(50): 8.2 μm, %D(90): 21.2 μm.

[0580] The microparticles were coated substantially as described in steps 1 to 4 of Comparative Example 10, except that:

[0581] - Before step 1, three Al cycles were performed;

[0582] - In step 1, 3 Zn + 1 Al cycles, repeated ten times per group (total 40 cycles per group);

[0583] - In step 3, steps 1 to 2 are repeated four times to form a second layer, a third layer, a fourth layer and a fifth layer of a mixed oxide having a zinc:aluminum atomic ratio of about 3:1;

[0584] - In step 4, three more Al cycles are performed to complete the process; and

[0585] - The ALD cycle is performed as follows (steps a to d represent the first cycle, subsequent cycles start from step a as specified in step e).

[0586] a. Reagent pulse 1:

[0587] i. Close the valve on the pipe between the pump and the ALD reactor.

[0588] ii. The valve on the diethylzinc or trimethylaluminum precursor bottle was opened for 1 second to allow the evaporated metal-containing precursor to fill the reaction chamber.

[0589] iii. The valve of the precursor bottle was closed and the chamber was left to stand for 30 s (soak time) before opening it to the pump again to ensure that the metal-containing precursor vapor reacted with the hydroxyl groups on the surface of the drug particles.

[0590] iv. The reactor was then pumped for 9 s.

[0591] v. Repeat steps a to d above 20 times.

[0592] b. Purge pulse: Purge the chamber with nitrogen in a continuous flow to remove unreacted reagents and organic gases.

[0593] c. Reagent pulse:

[0594] i. Close the valve on the pipe between the pump and the ALD reactor.

[0595] ii. Open the valve on the water precursor bottle for 1 s to allow the evaporated water to fill the reaction chamber.

[0596] iii. Close the valve of the water precursor bottle and let the chamber stand for 30 s (soak time) before opening the pump valve again to ensure that the water vapor adsorbs to the surface of the drug particles, presenting hydroxyl groups on the outside or particles.

[0597] iv. The reactor was then pumped for 9 s.

[0598] v. Repeat steps i to iv above 20 times.

[0599] d. Purge pulse: Purge the chamber with nitrogen in a continuous flow. Gaseous water and organic gases (in case this is not the first cycle) will be removed.

[0600] e. Repeat the cycle starting from step a.

[0601] Example 29

[0602] Determination of drug loading

[0603] This example describes the analysis of the liraglutide drug loading of the particles obtained in Example 28.

[0604] Materials and methods

[0605] To determine the drug loading (i.e., w / w% of liraglutide in the powder), HPLC (Prominence-i, Shimadzu, Japan) equipped with a diode array detector (Shimadzu, Japan) set at a wavelength of 220 nm was used using a 4.6×150 mm, 2.6 μm particle, PS C18 column (Kinetex, Phenomenex Inc., CA, USA). The material was dissolved in 2 M phosphoric acid in acetonitrile / water (1:1) and diluted with 0.1% trifluoroacetic acid acetonitrile / water (1:9) and further analyzed by HPLC (n=3).

[0606] The HPLC assay was set up according to Table 14.

[0607] Table 14. Parameters of the HPLC method.

[0608]

[0609] The concentration of the injected sample was automatically calculated by the software provided by the manufacturer. The drug loading was calculated according to the following formula, where C 样品 is the analytical concentration of liraglutide, V t is the total volume of the sample, m 样品 For the sample mass:

[0610]

[0611] result

[0612] The liraglutide drug loading of the particles according to Example 28 was determined to be 53.1%.

[0613] Example 30

[0614] Determination of coating integrity (DMSO)

[0615] This example describes the analysis of the coating integrity of the granules obtained in Example 28.

[0616] To determine coating integrity, a sample of the particles obtained in Example 28 was suspended in dimethyl sulfoxide (Rathburn, UK) at a concentration of 0.4 mg liraglutide per mL of solvent and rotated on an overhead stirrer for up to 72 hours. Intermittent sampling was performed, centrifuged at 2697 rcf for 10 minutes, and the supernatant was diluted with the above mobile phase A and then injected into the above HPLC system for quantification.

[0617] The concentration of the injected sample was automatically calculated by the software provided by the manufacturer. The amount of API dissolved in the coating integrity assay was calculated according to the following formula, where c a is the API concentration (mg / ml) obtained by analysis, V t is the total sample volume (ml), m API is the weighed API mass in (mg), and drug loading is the API content (fraction) in the coated API:

[0618] API dissolved amount (%) = 100·(c a ·V t ) / (m API Drug loading)

[0619] result

[0620] The amount of dissolved API in the coating integrity assay of the granules according to Example 28 was determined to be 3.0% after 3 hours and 5.2% after 72 hours.

[0621] Example 31

[0622] Preparation of formulations

[0623] This example describes the preparation of a formulation of coated lenalidomide particles as disclosed herein.

[0624] The powder of the coated microparticles of liraglutide according to the example was mixed with vet (Boehringer Ingelheim Animal Health, France) were mixed together. Vet is a veterinary medical product for animal injection, which contains 10 mg / mL of sodium hyaluronate in a sterile, isotonic phosphate buffer solution (pH 7.4). The composition of vet is presented in Table 15 below.

[0625] Table 15: Composition of vehicles used to suspend particles as disclosed herein.

[0626] Sodium Hyaluronate 10mg Active substances Sodium chloride 8.5mg Isotonicity Agents Sodium Acid Phosphate 0.223mg Buffer Sodium dihydrogen phosphate monohydrate 40 μg Buffer hydrochloric acid qs For pH adjustment Sodium hydroxide qs For pH adjustment Water for injection qsad 1mL Solvents

[0627] The microparticles were reconstituted by adding vehicle to the vial of powder containing the microparticles to obtain the following liraglutide concentrations:

[0628] - 10 mg / ml liraglutide according to the particles of Example 28.

[0629] Example 32

[0630] In vivo pharmacokinetic studies

[0631] This example describes a preclinical pharmacokinetic study of liraglutide following administration of a coated liraglutide formulation according to Example 31 in male Sprague Dawley rats.

[0632] Materials and methods

[0633] Male Sprague Dawley rats weighing approximately 300 g on the day of administration were provided by Charles River (UK). The duration of the study was 28 days. In each study group of the study, 4 rats were used. The study groups were the coated microparticle formulation according to Example 31, and as a control, a liraglutide solution ( Novo Nordisk, Denmark).

[0634] The hair of the application area was clipped before injection, and the injection site was marked. The suspension prepared with the coated liraglutide particles according to Example 28 as described in Example 32 was drawn into a 1 mL BD syringe, and a single subcutaneous injection (about 0.08 mL and 0.17 mL) was administered to the flank of each rat through a 23G needle (BD Microlance). In the control group, the control formulation (about 0.10 mL) was injected intravenously through the tail vein, and after a 7-day washout period, it was injected subcutaneously in the flank of each rat.

[0635] From animals to which the coated microparticles according to the examples have been administered, blood samples (about 0.2 mL) were collected from the jugular vein into K2EDTA tubes at the following time points: 1, 3, 6, 12, 24, 48, 72, 120, 168, 251, 384, 480, 576 and 672 hours after administration. Blood samples were collected at the following time points: 0.25, 1, 3, 6, 9, 12 and 24 hours after administration after intravenous injection of the control formulation, and at the following time points: 1, 2, 3, 6, 9, 12, 24 and 48 hours after subcutaneous injection of the control formulation.

[0636] Plasma was separated by centrifugation (1500 g, 4°C, for 10 min) as soon as possible after blood collection. The animals were sacrificed on the last day of the study. Plasma samples were stored in a -80°C freezer until analysis.

[0637] Plasma concentrations were determined using UPLC-MS / MS.

[0638] Samples were prepared by pipetting 35 μL of rat plasma into a 384-well plate using a TECAN EVO-2 liquid handling robot, adding 35 μL of 5% DMSO in acetonitrile, 70 μL of internal standard working solution. The 384-well plate was shaken for 15 minutes and centrifuged. All samples were then injected into a UPLC-MS / MS system (Xevo TQ-s micro coupled to an Acquity I-Class UPLC system, Waters, MA, USA) with the settings shown in Table 16 below.

[0639] Table 16. Parameters of the UPLC-MS / MS method.

[0640]

[0641] The pharmacokinetic analysis of lenalidomide in plasma was performed using the software Phoenix WinNonlin version 8.3 (Certara, USA) based on the non-compartmental analysis (NCA) used. The dose-normalized plasma concentrations of lenalidomide after a single subcutaneous administration of the formulation were assessed, and the following plasma pharmacokinetic parameters were evaluated:

[0642] ·C max – Maximum observed plasma concentration (ng / mL)

[0643] ·t max – Reach C max Time (h)

[0644] ·t last – Time (h) of last detection or last measurement (6-72h) of plasma concentration.

[0645] AUC 0-24h - Area under the curve (AUC) from time 0 to time 24 hours (h.ng / mL).

[0646] AUC last – From time 0 to t last AUC (h.ng / mL).

[0647] ·t 1 / 2,z –Terminal half-life (h)

[0648] AUC inf–AUC extrapolated to infinity (h.ng / mL).

[0649] ·F abs – Absolute bioavailability.

[0650] ·F rel –Relative bioavailability.

[0651] ·R 0-24h – The portion released 24 hours after injection.

[0652] Nominal plasma sampling time points and nominal doses were used for noncompartmental PK analysis. PK of the animals was calculated using the extravascular dose option in WinNonlin. Reported plasma concentrations below the LLOQ but above 1 ng / mL were included in the PK analysis, while unreported plasma concentrations below the LLOQ were omitted from the analysis.

[0653] C max and t max Derived from observed plasma concentration data. AUC was estimated by integrating the plasma concentration versus time curve and using linear interpolation for increasing plasma levels and logarithmic interpolation for decreasing plasma levels (increasing linear and decreasing logarithmic method). inf , using the concentration in the last quantifiable sample and λ z (the first order rate constant associated with the terminal portion of the curve) by dividing the area from AUC last Extrapolate to infinity. t 1 / 2,z By ln2 / λ z Calculate. abs Calculated as AUC from intravenous and subcutaneous injections inf The fraction of F, normalized by dose. rel Calculated as AUC from subcutaneous administration of control formulation and administration of coated microparticles according to Example 31 inf The fraction was normalized by dose.

[0654] result

[0655] Figure 7 The plasma concentration-time curves (liraglutide plasma concentration (ng / mL) versus sampling time days) of the sample obtained in Example 31 and the sample obtained using a known liraglutide solution as a control are shown.

[0656] More specifically, the change in plasma concentration (ng / mL) over time (days) is shown in a semi-logarithmic graph (y is a logarithmic scale). The solid squares show the results of rats administered with 5.6 mg / kg of coated microparticles of lenalidomide according to the example (n=4). The solid circles show the results of rats administered with 2.8 mg / kg of coated microparticles of lenalidomide according to the example (n=4). The hollow squares show the results of rats administered with 0.2 mg / kg intravenous control (liraglutide solution). The hollow circles show the results of rats injected with 0.2 mg / kg subcutaneous control (liraglutide solution).

[0657] Table 17. Results of evaluation of plasma pharmacokinetic parameters.

[0658]

[0659]

[0660] Example 33

[0661] Injectable 100mg / mL iHyonate

[0662] Preparation of formulations

[0663] The powder of the coated microparticles of liraglutide according to Example 28 was mixed with vet (Boehringer Ingelheim Animal Health, France) were mixed together.

[0664] The microparticles were reconstituted by adding vehicle to the vial of powder containing the microparticles to obtain the following liraglutide concentrations:

[0665] - 100 mg / ml liraglutide according to the granules of Example 28.

[0666] Materials and methods

[0667] The suspension prepared as described above with the coated liraglutide particles according to Example 28 was pipetted into 1 mL of HSW The three-part syringe (Henke-Sass, Wolf GmbH, Germany) was used. The syringe was equipped with a 27G×19mm injection needle (Microlance 3, BD, USA) and installed in a custom syringe holder in a TA.XTplus texture analyzer with Exponent Connect software (StableMicrosystems Ltd., UK). The texture analyzer was used to measure the force required to extend the piston when the syringe piston was pressed down to dispense the contents. In this method, the instrument was operated in compression test mode, moving its piston at a test speed of 10mm / s until a threshold force of 50N was measured, corresponding to the complete distribution of the syringe contents into the air. The threshold of 15N is considered to be the upper limit of acceptable force during injection.

[0668] result

[0669] The maximum injection force of the above suspension was determined to be 10.3N.

Claims

1. A method for preparing a pharmaceutical or veterinary composition in the form of a plurality of particles, the method comprising: (a) loading a plurality of solid cores comprising a bioactive agent into a stationary vapor deposition reactor chamber; as well as (b) applying a vapor deposition technique to surround, enclose and / or encapsulate the core with one or more layers comprising one or more coating materials, each of which comprises one or more metal-containing or metalloid-containing compounds; and (c) sequentially repeating step (b) as needed to form a plurality of particles having an average diameter based on weight, number and / or volume, the average diameter being between about 10 nm and about 100 μm, each particle comprising a corresponding solid core and a coating surrounding, enclosing and / or encapsulating the core, The vapor deposition technology includes: (1) introducing a pulse of a first reactant gas into the stationary vapor deposition reactor chamber and allowing the first reactant gas to contact the solid core for a predetermined soaking period of time; (2) after step (1), evacuating the fixed vapor deposition reactor chamber and / or purging the fixed vapor deposition reactor chamber with an inert gas; (3) introducing a pulse of a second reactant gas into the stationary vapor deposition reactor chamber and allowing the second reactant gas to contact the solid core for a predetermined soaking period of time; and (4) after step (3), evacuating the fixed vapor deposition reactor chamber and / or purging the fixed vapor deposition reactor chamber with an inert gas, The first reactant gas or the second reactant gas includes a metal-containing or metalloid-containing compound.

2. The method of claim 1 , wherein step (1), step (3), or both steps (1) and (3) comprise allowing the corresponding reactant gas to contact the solid core for a corresponding predetermined soaking period in the substantial absence of pumping that may cause gas flow.

3. A method according to claim 1 or claim 2, wherein step (1), step (3) or both steps (1) and (3) include allowing the corresponding reactant gas to contact the solid cores for a corresponding predetermined soaking time period in the absence of mechanical agitation of the plurality of solid cores.

4. The method of any one of the preceding claims, wherein the predetermined soaking period is from about 2 seconds to about 30 minutes.

5. The method according to any one of the preceding claims, wherein step (1), step (3) or both steps (1) and (3) comprise: o pumping the reactor for a predetermined pumping period after the predetermined soaking period; as well as o Repeating a predetermined number of the steps of introducing a pulse of a respective reactant gas, allowing the respective reactant gas to contact the solid core for a predetermined soaking period, and pumping the reactor for a predetermined pumping period.

6. A method according to any one of the preceding claims, wherein each repetition of step (b) is performed using the same or different first reactant gas and second reactant gas as the previous iteration of step (b).

7. A method according to any one of the preceding claims, wherein between one or more sequential pairs of step (b) iterations, the method comprises deagglomerating the coated solid core.

8. The method of claim 7, wherein one or more instances of depolymerizing the coated solid core comprises removing the coated solid core from the stationary vapor deposition reactor chamber, vibrating or sonicating the coated solid core, and reloading the coated solid core into the stationary vapor deposition reactor chamber to repeat step (b).

9. The method of claim 8, wherein the vibratory sieving or sonic screening of the coated solid cores is performed using a sieve having a mesh size determined so that the ratio of the size of the sieved or screened particles to the mesh size of the sieve is about 1:>1, preferably about 1:2, and optionally about 1:

4.

10. The method according to any one of the preceding claims, wherein at least 200 mg, optionally at least 1 g or at least 10 g of the solid core is loaded into the stationary vapour deposition reactor chamber for the vapour deposition technique to be applied.

11. A method according to any one of the preceding claims, wherein between 3 and 10 discrete layers of coating material are applied sequentially to the core.

12. The method of any one of the preceding claims, wherein the total thickness of the discrete layers of coating material is between about 0.5 nm and about 2 μm.

13. A method according to any of the preceding claims, wherein the maximum thickness of a single discrete layer of coating material is about 1 percent of the average diameter of the core based on weight, number or volume, including any other previously applied discrete layers of coating material located between the single discrete layer and the outer surface of the core.

14. The method according to any one of the preceding claims, wherein the coating material of one or more discrete layers comprises one or more inorganic coating materials.

15. The method of claim 14, wherein the one or more metal-containing or metalloid-containing compounds comprise hydroxides and / or oxides.

16. A method according to claim 14 or claim 15, wherein the one or more coating materials comprises silicon oxide, aluminum oxide, titanium dioxide, zinc sulfide and / or zinc oxide.

17. The method of claim 16, wherein the one or more coating materials comprise zinc oxide together with a mixture of one or the other or both of silicon dioxide and aluminum oxide.

18. A method according to any one of the preceding claims, comprising applying a separate layer of coating material to the core and / or previously coated core by atomic layer deposition.

19. A method according to any one of the preceding claims, wherein the core comprises a pharmaceutically acceptable excipient.

20. A method according to any preceding claim, wherein the carrier / excipient material is a sugar or sugar alcohol and / or is a pH adjusting agent.

21. The method of any one of claims 1 to 18, wherein the core consists essentially of the bioactive agent.

22. The method of any one of the preceding claims, wherein the bioactive agent is selected from the group consisting of analgesics, anesthetics, anti-ADHD agents, appetite suppressants, anti-addiction agents, antibacterial agents, antimicrobial agents, antifungal agents, antiviral agents, antiparasitic agents, antiprotozoal agents, anthelmintics, ectoparasiticides, vaccines, anticancer agents, antimetabolites, alkylating agents, antineoplastic agents, topoisomerases, immunomodulators, immunostimulants, immunosuppressants, anabolic steroids, anticoagulants, antiplatelet agents, anticonvulsants, antidementia agents, antidepressants, antidotes, antihyperlipidemic agents, antigout agents, antimalarials, antimigraine agents, anti-inflammatory agents, antiparkinsonian agents, antipruritics, antipsoriatic agents, antiemetics, antiobesity agents, anthelmintics, antiasthmatic agents, antibiotics , antidiabetic agents, antiepileptic agents, antifibrinolytic agents, antihemorrhagic agents, antihistamines, antitussives, antihypertensives, antimuscarinics, antimycobacterial agents, antioxidants, antipsychotics, antipyretics, antirheumatic agents, antiarrhythmic agents, antianxiety agents, aphrodisiacs, cardiac glycosides, cardiotonic agents, religious hallucinogens, reassuring drugs, euphoric drugs, appetite stimulants, antithyroid agents, antianxiety sedatives, hypnotic drugs, tranquilizers, astringents, antibacterial agents, beta blockers, calcium channel blockers, ACE inhibitors, angiotensin II receptor antagonists, renin inhibitors, beta-adrenergic receptor blockers, blood products, blood substitutes, bronchodilators, myocardial inotropes, chemotherapeutic agents, coagulants, corticosteroids, antitussives, diuretics, delirium drugs, antidepressants expectorants, fertility drugs, sex hormones, mood stabilizers, mucolytics, neuroprotectants, nootropics, neurotoxins, dopaminergics, free radical scavengers, growth factors, fibrates, bile acid sequestrants, cicatrizing agents, glucocorticoids, mineralocorticoids, hemostatics, hallucinogens, hypothalamic-pituitary hormones, immunizing agents, laxatives, antidiarrheals, lipid regulators, muscle relaxants, parasympathomimetics, parathyroid calcitonin, serenic, statins, stimulants, wakefulness agents, decongestants, dietary minerals, bisphosphonates, cough suppressants, ophthalmics, ontologicals, H1 antagonists, H2 antagonists, proton pump inhibitors, prostaglandins, radiopharmaceuticals, hormones, sedatives, anti Allergy-fighting agents, appetite stimulants, steroids, sympathomimetics, thrombolytics, thyroid agents, vasodilators, xanthines, erectile dysfunction improvers, gastrointestinal agents, histamine receptor antagonists, keratolytics, antianginal agents, nonsteroidal anti-inflammatory agents, COX-2 inhibitors, leukotriene inhibitors, macrolides, NSAIDs, nutritional agents, opioid analgesics, opioid antagonists, potassium channel activators, protease inhibitors, anti-osteoporosis agents, cognitive enhancers, anti-urinary incontinence agents, nutritional oils, anti-benign prostatic hypertrophy agents, essential fatty acids, non-essential fatty acids, cytokines, peptidomimetics, peptides, proteins, radiopharmaceuticals, anti-aging therapeutics, toxoids, serum, antibodies, nucleosides, nucleotides, vitamins, partial genetic material, nucleic acids,or any mixture of these drugs.

23. The method of any one of claims 1 to 21, wherein the bioactive agent is an anti-cancer agent.

24. The method of claim 23, wherein the bioactive agent is azacitidine.

25. A composition obtainable by means of a method according to any one of the preceding claims.

26. A pharmaceutical or veterinary preparation comprising: the composition according to claim 25; and a pharmaceutically acceptable or veterinarily acceptable adjuvant, diluent or carrier.

27. The pharmaceutical or veterinary preparation according to claim 26, which is in the form of a sterile injection and / or infusion dosage form.

28. A pharmaceutical or veterinary formulation according to claim 26 or claim 27 in the form of a liquid, sol or gel administrable via a surgical administration device forming a depot formulation.

29. A process for the preparation of a pharmaceutical or veterinary formulation as defined in any one of claims 26 to 28, said process comprising admixing a composition as defined in claim 25 with an associated pharmaceutically acceptable or veterinary acceptable adjuvant, diluent or carrier.

30. A composition according to claim 25 or a formulation according to any one of claims 26 to 28 for use in treating cancer, wherein the bioactive agent is as defined in claim 23 or claim 24.

31. Use of a composition according to claim 25 or a formulation according to any one of claims 26 to 28 for the manufacture of a medicament for treating cancer, wherein the bioactive agent is as defined in claim 23 or claim 24.

32. A method of treating cancer comprising administering to a patient in need of such treatment a composition according to claim 25 or a formulation according to any one of claims 26 to 28, wherein the bioactive agent is as defined in claim 22.

33. The composition or formulation for use according to claim 30, the use according to claim 31 or the method according to claim 32, wherein the bioactive agent is as defined in claim 24 and the cancer is myelodysplastic syndrome or one or more of its subtypes.

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