Field of the Invention

This invention relates to new pharmaceutical compositions, their use as medicaments and particularly to their administration to the lung to treat, for example, interstitial lung diseases.

Background and Prior Art

Interstitial lung diseases (ILDs) are a group of lung diseases that affect the interstitium, characterised by tissue around alveoli becoming scarred and/or thickened, and so inhibiting the respiratory process.

ILDs are distinct from obstructive airway diseases (e.g. chronic obstructive airway disease (COPD) and asthma), which are typically characterized by narrowing (obstruction) of bronchi and/or bronchioles. ILDs may be caused by injury to the lungs, which triggers an abnormal healing response but, in some cases, these diseases have no known cause. ILDs can be triggered by chemicals (silicosis, asbestosis, certain drugs), infection (e.g. pneumonia) or other diseases (e.g. rheumatoid arthritis, systemic sclerosis, myositis, hypersensitivity pneumonitis or systemic lupus erythematosus).

The most common ILDs are idiopathic pulmonary fibrosis (IPF) and sarcoidosis, both of which are characterised by chronic inflammation and reduced lung function.

Sarcoidosis is a disease of unknown cause that is characterised by collections of inflammatory cells that form lumps (granulomas), often beginning in the lungs (as well as the skin and/or lymph nodes, although any organ can be affected). When sarcoidosis affects the lungs, symptoms include coughing, wheezing, shortness of breath, and/or chest pain.

Treatments for sarcoidosis are patient-specific. In most cases, symptomatic treatment with non-steroidal anti-inflammatory drugs (NSAIDs) is possible, but for those presenting lung symptoms, glucocorticoids (e.g. prednisone or prednisolone), antimetabolites and/or monoclonal anti-tumor necrosis factor antibodies are often employed.

IPF on the other hand is a chronic lung disease characterized by a progressive and irreversible decline in lung function caused by scarring of the lungs. Symptoms typically include cough and shortness of breath. Although less prevalent than asthma and COPD, mortality rates from IPF are much higher (e.g. 5 times higher than that of asthma, despite asthma being 100 times more prevalent).

Current treatment of IPF includes oxygen supplementation. Medications that are used include pirfenidone or nintedanib, but with only limited success in slowing the progression of the disease. Further, both of these drugs commonly cause (predominantly gastrointestinal) side-effects.

IPF affects about 5 million people globally. Average life expectancy after diagnosis is around four years.

There are drawbacks associated with all of the aforementioned ILD drug treatments and there is a real clinical need for safer and/or more effective treatments.

Immunomodulatory imide drugs (IMIDs) are a class of immunomodulatory drugs that contain an imide group. The drug class includes thalidomide and analogues thereof, such as lenalidomide and pomalidomide.

Primary medical uses of IMIDs include the treatment of cancers, such as multiple myeloma and myelodysplastic syndrome (a precursor condition to acute myeloid leukaemia), as well as certain autoimmune diseases (including erythema nodosum leprosum, a painful vasculitic complication of leprosy). Off- label uses include other forms of cancer, such as Hodgkin's lymphoma and prostate cancer, as well as other conditions like primary myelofibrosis. Cyclodextrin-based thalidomide formulations for the local treatment of nosebleeds in hereditary hemorrhagic telangiectasia are also known (see Colombo et al, Int. J. Pharm., 514, 229 (2016)). Thalidomide's infamous history of causing birth defects following its use as an antiemetic during pregnancy is well known.

In addition to the above, there are published reports of thalidomide's potential use as a systemic treatment of ILDs, such as IPF, including IPF cough (see Zhao et al, Clin. Exp. Immunol., 157, 310 (2000), Zhang and Yang, Journal of Xi'an Jiatong University (Medical Sciences), 33, 622 (2012), Horton et al, Ann. Intern. Med., 157, 398 (2012) and Haraf et al, Am. J. Ther., 1 (2017)), bleomycin-induced pulmonary fibrosis (PF) in animals (Tabata et al, J. Immunol., 179, 708 (2007) and Don etal, Am. J. Transl. Res., 9, 4390 (2017)), and pulmonary sarcoidosis (Carlesimo et al, J. Am. Acad. Dermatol., 32, 866 (1995), Fazzi et al, Biomedicin & Pharmacotherapy, 66, 300 (2012) and Ye et al, Eur. Respir. J., 28, 824 (2006)). Lenalinomide has also been suggested for use systemically in pulmonary sarcoidosis (Jafari etal, Chest, 148, e35 (2015)).

There is no clear and direct disclosure or suggestion of the possibility of administering IMIDs by the pulmonary route for any indication, let alone in the specific, topical treatment of ILDs.

Particles comprising nanoporous (mesoporous) silica materials have been disclosed for use in general pharmaceutical and cosmetic applications in inter alia international patent application WO 2012/035074. Here, poorly soluble active ingredients are incorporated within nanopore channels of the silica particles. The use of similar particles with a specific particle size distribution for delivery of active ingredients to the respiratory tract are disclosed in international patent application WO 2018/202818.

Formulations comprising lenalidomide in various porous carriers including silica have been described in WO 2016/097030. In Ao et al ( Braz . J. Med. Res., 51, 1 (2018)), low density lipopeptide modified silica nanoparticles loaded with docetaxel and thalidomide for use in chemotherapy of liver cancer are disclosed. There is no suggestion in any of these documents of the use of such compositions in delivery of active ingredients to the lungs. See also US Patent application US 2007/0281036 Al. The use of porous materials for potential delivery of active ingredients to the lung has been disclosed. For example, US patents US 6,254,854, US 6,740,310 and US 7,435,408 all disclose polymeric materials, such as polyanhydrides and copolymers poly(lactic acid) grafted to amino acids; and international patent application WO 03/043586 discloses polymeric nanoparticles. Tartula et al ( J . Drug Target., 19, 900 (2011)), Li et al ( Nanomedicine , 11, 1377 (2015) and Wang et al, Nanoscale Research Letters, 12, 66 (2017)) disclose mesoporous silica nanoparticles for inhaled delivery of drugs. Finally, international patent application WO 03/011251 discloses mesoporous silicon carriers for pulmonary delivery.

Inhalation devices that are typically employed to administer active compounds to the lung include metered dose inhalers (MDIs), dry powder inhalers (DPIs) and soft mist inhalers (SMIs). DPIs may be divided into low, medium and high resistant DPIs.

The efficiency of DPIs, for example, is affected by two main forces 1) an inspiration air flow (IAF), which depends on a flow generated by the patient, and 2) a turbulence produced by the device.

A balance between these two forces is important for optimal performance of a device. If the IAF is too low, most of the drug is lost in the upper lung, i.e. the throat and the trachea. On the other hand, with most DPI-administered formulations, if the IAF is too high, more drug may be delivered in the lower lung (the bronchi and alveoli), but in a manner where there is often poor disaggregation of particles, and therefore dispersion of the drug in the lung.

Typical fixed-dose drug combinations for pulmonary delivery require powder homogeneity to deliver a uniform dose of drug to patients. This is often attempted by a simple blend of micronized drugs with coarse carrier particles.

In a MDI, the pharmaceutical composition is typically present in a liquid form, as a solution or suspension in a propellant, such as a hydrocarbon, a fluorocarbon or a hydrogen-containing fluorocarbon. In such systems it is often difficult to prevent dissolution of a bioactive compound from the particle or to prevent leakage of the compound from the drug-containing particle. Typically, solvents and/or surfactants are employed with a view to imparting stability to the suspension of drug particles. The compound needs to have a low solubility in the solvents that are used.

International patent application WO 2019/211624 describes how, in attempting to formulate specific mesoporous silica materials with specific properties with a view to inclusion of IMIDs, such as thalidomide, for pulmonary delivery, an undesirable agglomeration resulted, which could not be alleviated with conventional lubricant excipients. Surprisingly, incorporation of IMIDs, such as thalidomide, into the resultant silica particles resulted in deagglomeration of the resultant loaded particles, which was not expected. This rendered the resultant loaded silica particles of potential utility in the pulmonary delivery of such active ingredients.

We have also found that IMIDs show an excellent solubility profile when presented in this way, which renders such compositions of potential utility in the topical treatment of ILDs, such as IPF, by pulmonary administration.

By changing the process described in the above-mentioned patent application by functionalising the silica particles to include covalently-bonded alkyl groups using an appropriate functionalising agent, prior to loading the functionalised silica particles with IMID, it is possible to prolong the release of IMID from such particles.

Disclosure of the Invention

According to the invention, there is provided a pharmaceutical composition suitable for administration to the lung, which composition comprises a plurality of amorphous nanoporous (mesoporous) silica particles, in which one or more IMID is loaded into the pores of said particles, and wherein the silica particles have:

(a) a mass median aerodynamic diameter (MMAD), and/or a mean (or absolute) particle size, that is/are between about 0.1 pm and about 15 pm (e.g. about 10 pm); and

(b) a geometric standard deviation (GSD) that is less than about 4; and

(c) been surface modified, prior to loading with the one or more IMID, by functionalization and/or derivatization of free silanol groups with at least one organic moiety, which may be achieved by chemical reaction of those free silanol groups with a reagent that provides said at least one organic group, which compositions are hereinafter referred to as 'compositions of the invention'.

The loaded silica particles of the compositions of the invention may also have a mass density that is less than about 0.6 g/cm ³ , such as about 0.4 g/cm ³ , for example between about 0.15 and about 0.35 g/cm ³ .

In compositions of the invention, mean particle sizes may be presented as weight-, number-, or volume-, based mean diameters. As used herein, the term 'weight based mean diameter' will be understood by the skilled person to include that the average particle size is characterised and defined from a particle size distribution by weight, i.e. a distribution where the existing fraction (relative amount) in each size class is defined as the weight fraction, as obtained by e.g. sieving (e.g. wet sieving). The term 'volume based mean diameter' is similar in its meaning to weight based mean diameter, but will be understood by the skilled person to include that the average particle size is characterised and defined from a particle size distribution by volume, i.e. a distribution where the existing fraction (relative amount) in each size class is defined as the volume fraction, as measured by e.g. laser diffraction. As used herein, the term 'number based mean diameter' will be understood by the skilled person to include that the average particle size is characterised and defined from a particle size distribution by number, i.e. a distribution where the existing fraction (relative amount) in each size class is defined as the number fraction, as measured by e.g. microscopy. Other instruments that are well known in the field may be employed to measure particle size, such as equipment sold by e.g. Malvern Instruments, Ltd (Worcestershire, UK), Sympatec GmbH (Clausthal- Zellerfeld, Germany) and Shimadzu (Kyoto, Japan).

Mass median aerodynamic diameter (MMAD) will be understood by those skilled in the art to mean the diameter at which 50% of the particles by mass are larger and 50% are smaller over the total delivered dose as determined by any approved device, usually a cascade impactor such as a NGI, Andersen or Marple Miller impactor (see e.g. US Pharmacopeia at <601>; and/or www.uspbpep.com/usp31/v31261/usp31nf26sl_c601.asp). MMAD may be readily determined by those skilled in the art, for example by plotting on log probability paper the percentages of mass that is less than the stated aerodynamic diameters versus the aerodynamic diameters. The MMAD is taken as the intersection of the line with the 50% cumulative percent.

Particle sizes and/or MMADs of the particles may be varied depending on the preferred and/or intended site of delivery of the IMID compound. Particle sizes and/or MMADs that may be mentioned is between about 5 pm and about 15 pm. However, it is preferred that the particle size and/or MMAD of particles in compositions of the invention is between about 0.5 pm and about 8 pm, such as between about 1 pm and about 7 pm, for example between about 2 pm and about 6 pm, more preferably between about 3 pm and about 5 pm. This will mean that particles will tend to deposit primarily in the bronchioli.

GSD will be understood by those skilled in the art to be a measure of the spread of an aerodynamic particle size distribution. It is typically calculated as follows as:

(dgo/dio) ^1/2 wherein d9o and dio represent the diameters at which 90% and 10%, respectively, of the aerosol mass are contained, in diameters less than these diameters.

It is preferred that the GSD of particles in compositions of the invention is less than about 2.5, such as less than about 2.2, e.g. less than about 2.0, including less than about 1.8, or more preferably less than about 1.5, such as between about 1 and about 1.5.

Other parameters that may be used to define particles include mass density and the fine particle fraction (FPF). The FPF is the proportion of particles that have a diameter below about 5 pm. Preferred FPFs are at least about 50%, including at least about 60%, such as at least about 75% (e.g. at least about 80%), including at least about 85%, e.g. at least about 90%, such as at least about 95%, at least about 98%, and up to at least about 99%, at least about 99.9% or about 100%.

The silica particles may be manufactured by one or more of the processes described herein to a specification that has the MMAD and/or GSD (as well as other parameters) within any of the ranges or limits described herein. Alternatively, or preferably, the silica particles may be manufactured and thereafter separated and classified into the desired particle size range by any process known to those skilled in the art. For example, particles may be separated using cyclonic separation, by way of an air classifier, by elutriation, sedimentation and/or by sieving using one or more sieves or filters to obtain particles within a desired size range.

The silica particles that are employed in compositions of the invention may have a pore size that is between about 1 nm (e.g. about 2 nm) and about 100 nm (e.g. about 50 nm). Porous silica particles of compositions of the invention preferably have an average pore size that is in the range of about 2 nm (e.g. about 3 nm, such as about 4 nm, including about 5 nm and about 8 nm) up to about 30 nm (e.g. about 20 nm, such as about 16 nm (e.g. about 15 nm), including about 13 nm, such as about 12 nm (e.g. about 10 nm). Specific average pore sizes that may be mentioned include about 4.5 nm, about 5.0 nm, about 5.5 nm, about 6.5 nm, about 7.0 nm, about 7.5 nm, about 8.0 nm, about 8.5 nm, about 9.0 nm, about 9.5 nm, about 10 nm, about 11 nm, about 12 nm, about 13 nm, or about 14 nm. Such particles may also possess a pore volume that is between about 0.05 cm ³ /g, such as about 0.08 cm ³ /g, including about 0.09 cm ³ /g (e.g. about 0.1 cm ³ /g, such as about 0.2 cm ³ /g, or about 2 cm ³ /g) and about 3 cm ³ /g, such as about 2.5 cm ³ /g, including about 2.0 cm ³ /g (e.g. about 1.5 cm ³ /g or about 1.0 cm ³ /g), and/or may preferably possess a surface area that is in the range of about 35 m ² /g, e.g. about 40 m ² /g or about 50 m ² /g (such as about 100 m ² /g, including about 150 m ² /g or about 200 m ² /g) up to about and about 1,200 m ² /g, such as about 450 m ² /g, including about 350 m ² /g, e.g. about 300 m ² /g. All of these parameters may be determined by routine techniques, such as nitrogen adsorption isotherm (Brunauer, Emmett and Teller (BET)), mercury inclusion, and Barrett, Joyner and Halenda (BJH), methods.

Shapes of the porous particles may be controlled by the process of manufacture. Shape may be important for the incorporation and dissolution of the IMID. Thus, although silica particles may potentially be any shape (e.g. gyroids, rods, fibres, pseudo-spheres, cylinders, core-shells) in compositions of the invention, they are preferably essentially spherical. By 'essentially spherical', we mean that they may possess an aspect ratio smaller than about 20, more preferably less than about 10, such as less than about 4, and especially less than about 2, and/or may possess a variation in radii (measured from the centre of gravity to the particle surface), in at least about 90% of the particles that is no more than about 20% of the average value, such as no more than about 10% of that value, for example no more than about 5% of that value.

Porous silica particles may be loaded with one or more IMIDs by any suitable process known to those skilled in the art. For example, particles may be loaded by way of a solvent evaporation technique, for example as described hereinafter, impregnation, for example using a melt, use of supercritical CO2, shear mixing, co-grinding, spray-drying or freeze-drying. Well known equipment, such as a fluidized bed may be used. A preferred technique is solvent evaporation.

Loading the IMID into the silica particles means that it is loaded into the nanopores of the particles. It is preferred that the pores of the silica particles are loaded such that between about 0.1 and about 60%, preferably up to about 50%, such as up to about 45%, including up to about 40%, such as up to about 35% or up to about 30%, including up to about 25% (e.g. about 20% or about 10%) of the total weight of the loaded particles is IMID and, optionally, other pharmaceutical excipients, diluents or additives. In the alternative, it is preferred that up to about 60%, including up to about 70%, or up to about 80%, such as up to about 90%, e.g. up to about 100% of the pores of the silica particles are loaded with IMID and, optionally, other pharmaceutical excipients, diluents or additives. The entire mass of IMID does not have to be loaded into the pores of the particles and may otherwise be attached to the surfaces of the particle.

The IMID may be presented within the pores of the particles of compositions of the invention in an essentially crystalline state. By 'essentially crystalline', we mean that the IMID is at least about 95%, such as at least about 98%, for example at least about 99%, e.g. at least about 99.5%, and preferably at least about 99.9%, such as at least about 99.99%, crystalline, which may be detected by standard techniques, such as PXRD.

Alternatively, the IMID may be presented within the pores of the particles of compositions of the invention in an essentially amorphous state. By 'essentially amorphous', we mean that the IMID is no more than about 5%, such as no more than about 2%, for example no more than about 1%, e.g. no more than about 0.5%, and preferably no more than about 0.1% crystalline, which, again, may be detected by standard techniques, such as PXRD.

Presenting IMID in a crystalline or in an amorphous state within the pores of the particles of compositions of the invention means that the latter are capable of delivering a consistent and/or uniform dose of IMID, which is independent of solubility, after administration to the lung. We have found that, by incorporating IMID into the pores of the particles of the compositions of the invention, the IMID may remain in the same physical state (e.g. crystalline or amorphous), during and after manufacture, under normal storage conditions, and during use.

By this, we include that the IMID compound can be stored in the form of a composition of the invention, optionally in admixture with pharmaceutically acceptable carriers, diluents or adjuvants, under normal storage conditions, with an insignificant degree of solid state transformation (e.g. crystallisation, recrystallisation, loss of crystallinity, solid state phase transition, hydration, dehydration, solvatisation or desolvatisation). In addition to this, the IMID compound may be stored in this form under normal storage conditions, with an insignificant degree of chemical degradation or decomposition.

Examples of 'normal storage conditions' include temperatures of between minus 80 and plus 50°C (preferably between 0 and 40°C and more preferably ambient temperature, such as between 15 and 30°C), pressures of between 0.1 and 2 bars (preferably atmospheric pressure), relative humidities of between 5 and 95% (preferably 10 to 60%), and/or exposure to 460 lux of UV/visible light, for prolonged periods (i.e. greater than or equal to six months). Under such conditions, IMID may be found to be less than about 15%, more preferably less than about 10%, and especially less than about 5%, solid-state and/or chemically transformed. The skilled person will appreciate that the above- mentioned upper and lower limits for temperature and pressure represent extremes of normal storage conditions, and that certain combinations of these extremes will not be experienced during normal storage (e.g. a temperature of 50°C and a pressure of 0.1 bar). Thus, the loading of IMIDs into the pores of the silica particles in accordance with the invention can physically stabilize the IMID in an essentially crystalline and/or or an essentially amorphous form, and prevents it from undergoing solid state transformation, such that the physio-chemical properties of the drug do not change over time.

We prefer that the amorphous porous silica particles are biodegradable mesoporous silica.

The term 'biodegradable' means that the silica particles are dissolvable. Accordingly, a preferred embodiment of the invention is that the silica is a synthetic amorphous silica.

In order to be completely dissolvable, the silica particles of the compositions of the invention must be amorphous and therefore entirely non-crystalline (and remain so under normal storage conditions as hereinbefore defined), by which we mean that no crystallinity is detectable by standard techniques, such as PXRD. This is especially important considering the indications in which the compositions of the invention are intended to be used, in which injury by crystalline silica or other agents may be one of the causes of ILDs, such as PF.

Amorphous silica is less sensitive to humidity when compared to dry crystalline powder compositions that are typically used in pulmonary delivery of active ingredients.

Amorphous silica particles may be manufactured by processes known in the art. In one embodiment, porous particles may be manufactured by cooperative self- assembly of silica species and organic templates such as cationic surfactants such as alkyltrimethylammonium templates with varying carbon chain lengths, and counterions such as cetyltrimethylammonium chloride (CTA+CI- or CTAC) or cetyltrimethylammonium bromide (CTA+Br- or CTAB), or non-ionic species such as diblock and triblock polymer species, such as copolymers of polyethylene oxide and polypropylene oxide for example Pluronic 123 surfactant.

The formation of mesoporous silica particles occurs following the hydrolysis and condensation of silica precursors which can include alkylsilicates such as tetraethylorthosilcate (TEOS) or tetramethylorthosilicate (TMOS) in solution or sodium silicate solution. The mesoporous silica particle size can be controlled by adding suitable additive agents, e.g. inorganic bases, alcohols including methanol, ethanol, propanol, and other organic solvents, such as acetone, which affect the hydrolysis and condensation of silica species.

Pore size may not only be influenced by hydrothermal treatment of the reaction mixture such as heating up to 100°C or even above and also with the addition of swelling agents in the form of organic oils and liquids that expand the surfactant micelle template, but also, after condensation of the silica matrix, removing the templating surfactant by calcination typically at temperatures from about 500°C to about 650°C, or alternatively from 650°C up to about 750°C, in each case for e.g. several hours. Calcination at the higher of the above two temperature ranges not only burns away the organic template resulting in a porous matrix of silica (which the lower of the above two temperature ranges will also achieve), but also creates particles with one or more of the smaller average pore sizes mentioned hereinbefore (e.g. about 2 nm to about 14 nm, about 3 nm to about 13 nm and/or about 4 nm to about 12 nm (e.g. about 10 nm)), pore volumes (e.g. about 0.05 cm ³ /g to about 2.5 cm ³ /g, including about 0.08 cm ³ /g (e.g. about 0.09 cm ³ /g) up to about 2.5 cm ³ /g, including about 2 cm ³ /g (e.g. about 1.5 cm ³ /g, such as 1.0 cm ³ /g)), and/or surface areas mentioned herein (e.g. about 35 m ² /g to about 450 m ² /g, and/or about 50 m ² /g (e.g. about 100 m ² /g, including about 200 m ² /g) and about 400 m ² /g, such as about 350 m ² /g)). The template may alternatively be removed by extraction and washing with suitable solvents such as organic solvents or acidic of basic solutions.

In another embodiment, the porous silica particles may be manufactured by a sol-gel method comprising a condensation reaction between a silica precursor solution, such as sodium silicate or an aqueous suspension of silica nanoparticles as an emulsion, in either case with a non-miscible organic solution (such as benzyl alcohol), an oil, or a liquid polymer. In this process, droplets are formed by, for example, stirring or spraying the solution. Gelation of the silica may be carried out by means of changing pH, which may be carried out during or after the condensation step, and/or evaporation of the aqueous phase. The porosity of the particles here are formed either by exclusion due to the presence of the non-miscible secondary phase or by the jamming of the silica nanoparticles during evaporation.

Such particles may further be treated by washing to remove the non-miscible secondary phase and heating to induce condensation of the silica matrix. Furthermore, the particles may be treated by calcination as hereinbefore described to strengthen the silica matrix.

In another embodiment, the porous particles may be manufactured as porous glass through a process of phase separation in borosilicate glasses (such as Si02-B203-Na20), followed by liquid extraction of one of the formed phases through the sol-gel process, or simply by sintering glass powder. During a thermal treatment, typically between 500°C and 760°C, an interpenetration structure is generated, which results from a spinodal decomposition of the sodium-rich borate phase and the silica phase.

The porous particles may also be manufactured using a fumed process. In this method, fumed silica is produced by burning silicon tetrachloride in an oxygen- hydrogen flame producing microscopic droplets of molten silica which fuse into amorphous silica particles in three-dimensional secondary particles which then agglomerate into tertiary particles. The resulting powder has an extremely low bulk density and high surface area.

Irrespective of how the silica particles are manufactured (e.g. as described herein or otherwise), following such manufacture and prior to loading with IMID, they are surface modified by chemical reaction of free silanol groups with a reagent that provides at least one organic group.

This can be achieved by surface modification of silica particles by reaction with an alkoxysilane, and/or an alkylhalosilane, many of which are commercially available, for example as described hereinafter. These reagents are capable of forming 1 to 3 Si-O-Si links to the surface by way of a condensation reaction with surface silanol groups.

Typical functionalising reagents that may be employed to achieve this include 3-aminopropyl triethoxysilane, 3-mercaptopropyl trimethoxysilane and various PEG-silanes. A functionalising reagent that may be mentioned is an alkylhalosilane, which alkylhalosilane may be an alkylchlorosilane containing up to 4 (e.g. 3) alkyl groups, such as between 1 and 4 (e.g. 3) linear or branched C 1-24 alkyl groups, such as Ci-is alkyl groups, including Ci-io alkyl groups, e.g. a di- or tri-C alkylchlorosilane, such as tripropylchlorosilane, triethylchlorosilane or trimethylchlorosilane.

Alternative (and/or preferred) functionalising reagents include reactive species that comprise only one alkyl chain, such as alkyl esters of halocarboxylic acids (e.g. alkylchloroformates) or haloalkanes, for example as described hereinafter. Preferred reagents include those containing linear or branched Ci- ₂₄ alkyl groups, such as Ci-is alkyl groups, including Ci-io alkyl groups, such as Ci-4 alkyl groups, e.g. methylchloroformate or methyliodide, propylchloroformate or propyl iodide or, more preferably, ethylchloroformate or ethyliodide.

Surface modifications of this type may be carried out by reacting said functionalising reagent with silica particles, optionally in the presence of an appropriate solvent (e.g. toluene or tetrahydrofuran) and/or an appropriate base (e.g. imidazole, N-methylmorpholine or sodium carbonate), for example as described hereinafter, or alternatively as described in Zhao and Li, J. Phys. Chem., 102, 1556 (1998), Taib etal, Int. J. Chem., 3, 2 (2011), or Chmielowiec and Morrow, J. Colloid Interface Sci., 94, 319 (1983).

However it is carried out, functionalisation of silica is preferably achieved with a low conversion (i.e. only functionalising a portion of the surface -OH groups). The choice of the functionalising reagent, solvent and temperature enables a functionalisation method to obtain the desired surface coverage with organic (e.g. alkyl) groups and therefore the desired properties.

Longer alkyl chains (e.g. ethyl groups and higher) are expected to lower static interactions between the silica particles, and be more hydrophilic. Further, because the obtained products are known to be sensitive to hydrolysis to liberate the corresponding alcohol, functionalization with ethyl groups is preferred as it will liberate only ethanol in the body.

After loading with IMID, the loaded silica particles may be admixed with one or more fatty acid- or lipid-based surfactants. Such admixing is preferably done by dry mixing said surfactant with said loaded particles, more preferably by way of a high energy mixing process. Appropriate high energy mixing equipment may include, for example, intensive mechanical processors (e.g. the Nobilita- 130 Unit Mechanofusion System (Hosokawa Micron Corporation, Osaka, Japan) or Laboratory Mixer Granulator P 1-6 (DIOSNA Dierks &. Sohne GmbH, Osnabruck, Germany)), for example under appropriate mixing conditions, such as those described hereinafter and/or in Zhou et al, J. Pharm. Sci., 99, 969 (2010).

Admixing may also be achieved by other techniques known to those skilled in the art, including spraying a solution or a suspension of said surfactant onto the surfaces of said particles by a suitable means, such as using a fluidized bed and/or a jet mill.

The term 'fatty acid- or lipid-based surfactant' will be understood to include any surfactant comprising a long (Cs-24) hydrocarbon chain. Surfactants comprising such hydrocarbon chains are or may be derived from oilseeds (e.g. palm, palm kernel, coconut, etc.), and may be saturated, branched, linear and/or aromatic. Surfactants based on lipids or fatty acids may be non-ionic, but are preferably ionic.

Ionic surfactants may include those with a cationic head group (e.g. primary, secondary, or tertiary amines; primary and secondary amines and quaternary ammonium salts); Zwitterionic (amphoteric) surfactants (e.g. sultaines, betaines and phospholipids); but more preferably include anionic surfactants, such as salts of sulfate esters (e.g. ammonium lauryl sulfate and sodium lauryl sulfate), sulfonate esters and phosphate esters or, more preferably, carboxylate esters.

Anionic surfactants based on carboxylate esters include carboxylate salts (soaps), which surfactants comprises an alkali, or an alkaline earth, metal ion (e.g. sodium, potassium, calcium or magnesium) and one or more fatty acid chain with at least 10, such as at least 12, including at least 14, such as at least 16, carbon atoms. Preferred specific anionic surfactants in this class include sodium stearate, sodium lauroyl sarcosinate and carboxylate-based fluorosurfactants, such as perfluorononanoate and perfluorooctanoate. However, we prefer that the surfactant that is employed in compositions of the invention is magnesium stearate.

The amount of surfactant that may be employed in compositions of the invention is in the range of about 0.1% to about 12% by weight of the composition, such as about 0.2% to about 11%. Preferred amounts are in the range of about 1%, such as about 2%, including about 3%, up to about 10% by weight of the composition. Specific amounts that may be included are thus about 0.5%, about 1.0%, about 1.5%, about 2.0%, about 2.5%, about 3.0%, about 3.5%, about 4.0%, about 4.5%, about 5.0%, about 5.5%, about 6.0%, about 6.5%, about 7.0%, about 7.5%, about 8.0%, about 8.5%, about 9.0%, about 9.5%, about 10.0% and about 10.5%.

According to a further aspect of the invention there is a provided a process for the production of a composition of the invention, which process comprises:

(a) forming mesoporous silica particles by a method as described hereinbefore;

(b) optionally, calcining at a temperature of between about 500°C and about 600°C, or about 650°C and about 750°C, in order to provide particles having relevant pore sizes, pore volumes, and/or surface areas, within the ranges described herein;

(c) surface modifying said silica particles by chemical reaction of the particles and in particular the free surface silanol groups thereof with a reagent that provides at least one organic group, as described hereinbefore;

(d) if particles formed after any one of steps (a) to (c) do not, at least in part, have an MMAD, and/or a mean (or absolute) particle size, and/or a GSD within the ranges specified herein, separating particles after the relevant step as described herein so as to obtain particles having an MMAD, and/or a mean (or absolute) particle size, and/or a GSD within those ranges; and, finally,

(e) loading the obtained particles with an IMID as described herein.

The process described herein for production of compositions of the invention has the advantage that it allows the production of particles with sizes that enable better control of the site of deposition of the particles in the lung, so enabling accurate tailoring of site-specific lung delivery (e.g. improved delivery to the deep lung) compared to prior art inhalation formulations comprising other drugs. The process described herein also reduces manufacturing costs compared to processes in which separation is conducted after loading particles with a bioactive compound. This may improve the yield and efficiency of the manufacturing process. The process also provides for a higher drug loading of the bioactive compounds in final dosage forms comprising compositions of the invention.

In view of the particle size of the silica particles of the compositions of the invention, particle aggregation was expected. Aggregation of dry particles in the micron-sized range is a well-known phenomenon in particle and powder processing. Aggregation is caused by numerous attractive (ubiquitous) forces, such as van der Waals forces and/or electrostatic interactions. In many cases, particle aggregation causes unwanted problems such as poor handling and flowability and sticking to containers.

Particles within the size range mentioned herein are also often prone to aggregation in air due to the large surface area to volume ratio.

In view of the above, particle aggregation is a serious hurdle for pulmonary delivery, given that the particle size is critical to ensure correct distribution of the particles in the lung. Aggregation of particles would be expected to lead to accumulation in the throat and upper airways thereby limiting the effectiveness of the formulation. Additionally, aggregation of particles or sticking of particles in the capsules during inhalation is a severe limitation.

As described hereinafter, when small amounts of pre-loaded silica particles that might be considered to be suitable for inhalation were investigated, as expected, the particles aggregated and had poor flow properties. When attempts were made to use common excipients that are normally employed to reduce particle aggregation, such as lubricants, the flow properties of the particles did not improve.

However, as also described hereinafter, when the same particles were loaded with an IMID (e.g. thalidomide), the flow properties of the particles was dramatically improved without the addition of further excipients. Compositions of the invention are useful as medicaments/pharmaceuticals. Their unexpectedly good flow properties renders them suitable for pulmonary delivery.

In particular, compositions of the invention find particular utility in the pulmonary treatment of an ILD. In this respect, there is provided a composition of the invention for use in the treatment of an ILD by pulmonary administration as well as the use of a composition of the invention for the manufacture of a medicament for the treatment of an ILD by pulmonary administration.

ILDs include autoimmune diseases (in which the immune system attacks the body), such as lupus, rheumatoid arthritis, sarcoidosis and scleroderma/systemic scelorosis, lung inflammation due to breathing in a foreign substance such as dust, fungus or mould (hypersensitivity pneumonitis), side effects of medicines (such as nitrofurantoin, sulfonamides, bleomycin, amiodarone, methotrexate, gold, infliximab, etanercept, and other chemotherapy medicines) or radiation treatment to the chest, or occupational lung disease, brought on by working with or around asbestos, coal dust, cotton dust, and silica dust.

The term 'ILD' may in addition and/or in the alternative be understood by those skilled in the art to include any pulmonary condition characterized by an abnormal healing response, including chronic inflammation, reduced lung function and/or scarring, irrespective of the cause, such as sarcoidosis, and PF, especially IPF. The term may also include diseases and/or conditions that are known to lead to, and/or be causes of, such pulmonary conditions, such as systemic sclerosis. In this respect there is further provided a composition of the invention for use in the condition that leads to and/or is a cause of an ILD, such as PF or IPF, including systemic sclerosis.

Compositions of the invention may also be useful not only in treating underlying conditions, such as ILDs, but also treating, preventing and/or alleviating specific symptoms of those diseases, including wheezing, tiredness, weight loss/cachexia, chest pain and especially cough and shortness of breath. In this respect, 'treatment of an ILD' includes treatment of the symptoms of an ILD, such as IPF cough. According to a further aspect of the invention there is provided a method of treatment of an ILD, which method comprises the pulmonary administration of a pharmacologically-effective amount of an IMID in the form of a composition of the invention to a patient in need of such treatment.

In addition, compositions of the invention may also be useful in the treatment or prevention of any fibrotic condition of one or more internal organs characterised by the excessive accumulation of fibrous connective tissue, and/or in the treatment or prevention of fibrogenesis and the morbidity and mortality that may be associated therewith. Such fibrosis may be associated with an acute inflammatory condition, such as acute respiratory distress syndrome (ARDS), severe acute respiratory syndrome (SARS), and multiple- organ inflammation, injury and/or failure, which may be caused by internal or external trauma (e.g. injury), or by an infection.

Such conditions may thus result from sepsis or septic shock caused by a viral, bacterial or fungal infection. Furthermore, acute lung injury, ARDS and, particularly, SARS may be caused by viruses, such as coronaviruses, including the novel SARS coronavirus 2 (SARS-CoV-2), which may result in internal tissue damage and/or dysfunction of relevant internal (e.g. mucosal) tissues, such as the respiratory epithelium. Such tissue damage may in turn give rise to severe fibrosis. For example, the SARS disease caused by the novel coronavirus SARS- CoV-2 (coronavirus disease 2019 or COVID-19) is known in many cases to result in fibrosis.

In this respect there is further provided a composition of the invention for use in the treatment of a 'precursor' condition that may lead to fibrosis, and/or in the treatment of a fibrotic condition, such as an ILD (e.g. PF or IPF), that is caused by such a precursor condition. Such precursor conditions may include systemic sclerosis, acute injury or a relevant (e.g. viral) infection, including those mentioned above (such as SARS-CoV-2).

'Patients' include mammalian (particularly human) patients. Human patients include both adult patients as well as paedeatric patients, the latter including patients up to about 24 months of age, patients between about 2 to about 12 years of age, and patients between about 12 to about 16 years of age. Patients older than about 16 years of age may be considered adults for purposes of the present invention. These different patient populations may be given different doses of IMID.

IMIDs include lenalidomide, pomalidomide and, especially, thalidomide. IMIDs may be administered in the form of racemates, single enantiomers and/or pharmaceutically-acceptable salts.

Pharmaceutically-acceptable salts of IMIDs include base addition salts and preferably acid addition salts. Such salts may be formed by conventional means, for example by reaction of a free acid or, preferably, free base form of an active ingredient with one or more equivalents of an appropriate acid or base as appropriate, optionally in a solvent, or in a medium in which the salt is insoluble, followed by removal of said solvent, or said medium, using standard techniques (e.g. in vacuo, by freeze-drying or by filtration). Salts may also be prepared by exchanging a counter-ion of an active ingredient in the form of a salt with another counter-ion, for example using a suitable ion exchange resin.

However, it is preferred that IMIDs are employed in formulations of the invention in their free form (i.e. not in the form of a pharmaceutically- acceptable salt).

Pulmonary delivery means compositions of the invention are adapted for delivery to the lungs by direct inhalation, and thereby giving rise to the direct topical treatment by IMIDs of ILDs in the lungs.

Administration of IMID is preferably intermittent. The mode of administration may also be determined by the timing and frequency of administration, but is also dependent, in the case of the treatment of ILDs, on the severity of the condition, or otherwise on the need for treatment.

Compositions of the invention may also impart, or may be modified to impart, an immediate, or a modified, release of IMIDs.

Compositions of the invention may be combined with other excipients that are well known to those skilled in the art for pulmonary delivery of active ingredients. For example, optional excipients may include propellants; surfactants, such as poloxamers; sugars or sugar alcohols, such as lactose, glucose, mannitol or trehalose; lipids, such as DPPC, DSPC, DMPC, cholesterol; amino acids, such as leucine or trileucine; cyclodextrins, hydroxypropylated chitosan, poly-lactic-co-glycolic acid (PLGA); antioxidants; humidity regulators and the like, though such are by no means essential. Indeed, we have found that, in the pulmonary delivery of compositions of the invention, fewer additional excipients are needed, which may reduce cost of manufacture.

Inhalation devices that may be employed to administer compositions of the invention to the lung include MDIs, SMIs and DPIs, including low, medium and high resistant DPIs.

Compositions of the invention may form stable compound suspensions when suspended in solvents that are typically employed in MDIs. The loaded silica particles may be well-dispersed in different solvents and may be further modified to prevent dissolution or leakage of drug into the solvent before delivery to the target site or lung.

In view of the fact that, as mentioned hereinbefore, compositions of the invention have unexpectedly good flow properties, this minimizes the need for disaggregation of the particles by increased IAF and turbulence produced by the inhalation device. This in turn improves the balance between the two forces discussed hereinbefore, and thus improves delivery of IMID to the lower lung without loss of drug in the upper lung. This further reduces the dependence on the inhalation device that is employed.

According to a further aspect of the invention, there is further provided a drug delivery device adapted for delivery of active ingredients to the lung, which delivery device comprises a composition of the invention.

The delivery device may be a MDI, a DPI or a SMI. When used in, in particular, a MDI, the composition of the invention is optionally mixed with a propellant, which propellant has a sufficient vapour pressure to form aerosols upon activation of the delivery device. The propellant may be selected from the group a hydrocarbon, a fluorocarbon, a hydrogen-containing fluorocarbon and a mixture thereof. The above-mentioned excipients may be commercially-available or otherwise are described in the literature, for example, Remington The Science and Practice of Pharmacy, 19th ed., Mack Printing Company, Easton, Pennsylvania (1995) and the documents referred to therein, the relevant disclosures in all of which documents are hereby incorporated by reference. Otherwise, the preparation of suitable pulmonary formulations may be achieved non-inventively by the skilled person using routine techniques.

Similarly, the amount of IMID in the formulation will depend on the severity of the condition, and on the patient, to be treated, but may be determined by the skilled person.

For example, suitable lower daily doses (calculated as the free base) of thalidomide in adult patients (average weight e.g. 70 kg), may be about 0.01 mg, such as about 0.1 mg, for example about 1 mg, or about 5 mg, per day. Suitable upper limits of daily dose ranges of e.g. thalidomide may be about 200 mg, such as about 50 mg, including about 25 mg, such as about 10 mg.

All of the above doses are calculated as the free base and, again, doses may be split into multiple individual doses per day. Inhaled doses may be given between once and six, such as four times daily, preferably three times daily and more preferably twice daily. Alternatively, inhaled doses may be given between once and four times weekly, for example every other day.

In any event, the medical practitioner, or other skilled person, will be able to determine routinely the actual dosage, which will be most suitable for an individual patient, depending on the severity of the condition and mode and frequency of administration.

The dose administered to a mammal, particularly a human, in the context of the present invention should be sufficient to affect an appropriate response (e.g. a reduction in symptoms such as, in the case of IPF, cough) in the mammal (e.g. human) over a reasonable timeframe (as described hereinbefore). One skilled in the art will recognize that the selection of the exact dose and composition and the most appropriate delivery regimen will also be influenced by inter alia the pharmacological properties of the formulation, the nature, stage and severity of the condition being treated, and the physical condition and mental acuity of the recipient, as well as the age, condition, body weight, sex and response of the patient to be treated, and the stage/severity of the disease, as well as genetic differences between patients.

The above-mentioned dosages are exemplary of the average case; there can, of course, be individual instances where higher or lower dosage ranges are merited, and such are within the scope of this invention.

Although intended for inhalation, compositions of the invention may be co administered with other pharmaceutical formulations comprising different (or the same) active ingredients that are intended for the treatment of ILDs (whether administered pulmonarily, orally, or otherwise). For example, uses and methods that involve pulmonary administration of compositions of the invention may be combined with one or more treatments comprising other active ingredients that are useful in the treatment of ILDs (or a peroral treatment comprising one or more IMID).

Relevant active ingredients for use in IPF include, for example, anti-fibrotics, such as nintedanib and pirfenidone; corticosteroids, such as cortisone, dexamethasone and prednisone; inflammation suppressants, such as cyclophosphamide; other immunosuppressants, such as azathioprine and mycophenolate mofetil; and antioxidants, such as N-acetylcysteine. Relevant active ingredients for use in sarcoidosis include, for example, corticosteroids, such as dexamethasone, cortisone, prednisone and prednisolone; antimetabolites; immune system suppressants, such as methotrexate, azathioprine, leflunomide, mycophenoic acid/mycophenolate mofetil, cyclophosphamide; aminoquinolines; monoclonal anti-tumor necrosis factor antibodies, such as infliximab and adalimumab; AT2 receptor agonists, such as C21 (N-butyloxycarbonyl-3-(4-imidazol-l-ylmethylphenyl)-5-iso-bu tylthio- phene-2-sulfonamide); TNF inhibitors, such as etanercept; and painkillers, such as ibuprofen and paracetamol; cough suppressants and/or expectorants.

Relevant patients may thus also (and/or may be already) be receiving such therapy for the treatment of their ILD based upon administration of one or more of such active ingredients, by which we mean receiving a prescribed dose of one or more of those active ingredients mentioned herein, prior to, in addition to, and/or following, treatment with IMID. Pharmaceutically-acceptable salts, and doses, of other active ingredients useful in the treatment of ILDs include those that are known in the art and described for the drugs in question to in the medical literature, such as Martindale - The Complete Drug Reference (35 ^th Edition) and the documents referred to therein, the relevant disclosures in all of which documents are hereby incorporated by reference.

Wherever the word 'about' is employed herein, for example in the context of absolute numbers (e.g. ages) and relative numbers (e.g. multiples), amounts, i.e. absolute amounts, such as sizes (aerodynamic diameters, particle sizes and pore sizes), doses, weights or concentrations of (e.g. active) ingredients, pore volumes, particle surface areas, particle densities, temperatures, patient populations or time periods or frequencies; or relative amounts including percentages, standard deviations and aspect ratios, it will be appreciated that such variables are approximate and as such may vary by ±10%, for example ±5% and preferably ±2% (e.g. ±1%) from the actual numbers specified. In this respect, the term 'about 10%' means e.g. ± 10% about the number 10, i.e. between 9% and 11%.

In addition to the advantages mentioned hereinbefore, compositions of the invention provide for an improved drug loading for the reasons described hereinbefore. This enables high quantities/doses of bioactive compound to be presented in dosage forms comprising compositions of the invention, and also efficient delivery of such higher doses to the desired site in the lung in a consistent/uniform manner. This in turn means that the frequency of dosing may be reduced, so increasing the effectiveness and efficiency of treatment as well as reducing costs of healthcare.

Furthermore, improved efficiency of deposition of the IMID compound in the lung in view of the low amount of aggregation of particulates within which the IMID is loaded allows for more efficient and more precise lung delivery, and thus an improved therapeutic effect.

Homogeneity in terms of both carrier particle size and drug distribution within the composition may also be improved by compositions of the invention. In addition, if desired, compositions of the invention may include additional bioactive compounds (IMID or otherwise as described hereinbefore), which may also be loaded into silica particles without substantial loss of material. This may be useful in e.g. co-therapy as described hereinbefore, and moreover may further reduce cost of manufacture.

Compositions of the invention also have the advantage that the dissolution kinetics of the IMID compound is largely independent of particle size, morphology of the compound and site of delivery in the lung. Adjusting pore size may thus be employed to tailor drug dissolution kinetics, but the dissolution kinetics of the drug will be independent of the position of the particles in the lung. Importantly, compositions of the invention may give rise to more prolonged release of IMID once particles are deposited in the lung.

The uses/methods described herein may otherwise have the advantage that, in the treatment of ILDs, they may be more convenient for the physician and/or patient than, be more efficacious than, be less toxic than, have a broader range of activity than, be more potent than, produce fewer side effects than, or that it may have other useful pharmacological properties over, similar methods (treatments) known in the prior art, whether for the treatment of ILDs or otherwise.

The invention is illustrated, but in no way limited, by the following examples, in which Figure 1 shows in vitro release profiles for thalidomide from silica particles in simulated lung fluid.

Examples

Comparative Example 1 Silica Particle Manufacture I

Pluronic 123 (triblock co-polymer, E020P070E020, Sigma-Aldrich; 4 g; templating agent) and 1,3,5-trimethylbenzene (TMB; mesitylene, Sigma- Aldrich; 3.3 g; swelling agent) were dissolved in 127 mL of distilled H2O and 20 mL of hydrochloric acid (HCI, 37%, Sigma-Aldrich) while stirring at room temperature for 3 days. The solution was preheated to 40°C before adding 9.14 mL of tetraethyl orthosilicate (TEOS; Sigma-Aldrich). The mixture was stirred for another 10 minutes at a speed of 500 rpm, then kept at 40°C for 24 hours, and then hydrothermally treated in the oven at 100°C for another 24 hours. Finally, the mixture was filtered, washed and dried at room temperature.

The product was calcined to remove the surfactant template and swelling agent. The calcination was conducted by heating to 600°C with a heating rate of 1.5°C/min and kept at 600°C for 6 hours, followed by cooling to ambient conditions. The resultant product was a white powder comprising porous silica particles.

Comparative Example 2 Silica Particle Manufacture II

A dispersion (14 wt%) of silica nanoparticles (10 nm) in water (pH 9) (400 mL) was poured into benzyl alcohol (800 mL) warmed to 50°C and stirred at 300 rpm with an overhead stirrer (Silverson, UK) for 20 minutes.

A drop of acetic acid was added and vacuum (200 bar) was applied during heating at 80°C to remove the aqueous phase. The resulting particles were collected by filtration and washing with acetone.

The product was calcined by heating to 600°C with a heating rate of 1.5°C/min and kept at 600°C for 6 hours, followed by cooling to ambient conditions. The resultant product was a white powder comprising porous silica particles in the size range 2 to 4 microns measured by scanning electron microscope (JEOL, Japan) and by electrical sensing zone method (Elzone, Micromeretics USA). The particles were further treated by refluxing in ammonium hydroxide overnight followed by filtering and refluxing in nitric acid overnight and finally filtered and washed in water and oven dried at 80°C.

Comparative Example 3 Silica Particle Separation

100 g of nanoporous silica particles (preparable and/or prepared as described in Comparative Example 1 and/or Comparative Example 2 above) were fed into an air classifier (TTS, Hosokawa-Alpine), with the air flow adjusted from 53 to 42 m ³ /h and the speed set at between 2,475 and 13,500 rpm. 11 g of fines and 8 g of course materials were collected. The particle size distribution calculated as (Dgo/Dio) was reduced from 4.5 to 1.8.

Comparative Example 4 Nanoporous Silica Particle Properties

Two porous silica particle types with different porosities and densities (1.2 mL/g and 0.9 mL/g, and 0.18 mg/mL and 0.33 mg/mL, respectively) were prepared essentially as described in Comparative Example 1 and/or Comparative Example 2, and were characterised to determine their MMAD, GSD and GPS (mean particle size) using an 8-Stage Cascade Impactor (Marple), as shown in Table 1 below.

Table 1

By optical observation, it could be seen that, in small quantities, the above particles aggregated and had poor flow properties.

Attempts were made to reduce particle aggregation for formulation by using the well-known glidant magnesium stearate. This is an approved excipient for inhalation products. The particles were mechanically mixed with magnesium stearate (Sigma) at several different weight ratios in the range 1-5% magnesium stearate, but the flow properties of the particles did not improve.

Comparative Example 5

Loading of Thalidomide in Silica Particles

Thalidomide was encapsulated into the porous silica particles of Comparative Example 4 above (those with the bulk density of 0.18 mg/mL and the MMAD of 4.33 mhi) by a solvent impregnation and evaporation method. A concentrated solution of thalidomide was made in a chosen good solvent for the drug, and various known masses of nanoporous silica particles were added to the solution. The solvent was removed by evaporation.

In one example, thalidomide (200 mg; Sigma) was dissolved in methylene chloride (120 mL; Sigma) at room temperature in a round-bottomed flask. Nanoporous silica particles (300 mg) were added to the thalidomide solution.

The mixture was stirred for 30 minutes at 40°C. The solvent was evaporated with controlled evaporation under a reduced pressure of 200 mBar in a rotary evaporator, with a water bath temperature of 60°C. The resultant dry powder that was collected was free flowing. The samples were further dried at 40°C under vacuum for 4 hours.

The samples were characterized by TGA to evaluate drug loading. Loading amounts of 20, 40 and 50 wt% (calculated as mass of drug/mass of loaded particles) were determined.

The physical state of the drug (crystalline vs amorphous) was measured with DSC. Thalidomide has a melting peak in DSC of around 270°C. Thalidomide was stabilized in an amorphous state in the samples with a drug loading up to 40%. There was a small melting peak observed with the samples with a drug loading amount of 50% indicating presence of crystalline drug. This demonstrates that the maximum loading capacity of the particles was probably reached at 50% loading i.e. complete loading of the particles and it is likely that crystalline drug resided on the outside of the particles.

Analysis by light microscopy showed that the free drug was fully encapsulated in the porous silica particles.

Comparative Example 6

Loading of Silica Loaded with 40% ibv weight) Thalidomide into Capsules

Two Sincapsules® containing 20 mg of sodium cromoglycate (Sanofi AB) were opened and emptied using pressurized air. One of the capsules was filled with 2.5 mg of silica particles loaded with 40% (wt%) of thalidomide, obtained as described in Comparative Example 5 above and the other was loaded with the same amount of unloaded silica particles as a control.

The Sincapsules were inspected before use. It was clear that unloaded silica particles were sticking to the capsule wall. Surprisingly, silica particles loaded with 40% (weight) thalidomide were nonadherent and moved around freely when moving the capsule. Thus, the loading of thalidomide into the silica particles improved the flow properties of the particles without the addition of further excipients.

The closed capsules were loaded in an Intal® Spinhaler® according to the manufacturer's instructions in the packaging insert. The full doses were inhaled by one healthy volunteer, following the instructions in the packaging insert, making sure that the full content was inhaled.

Example 1

Manufacture of a Composition of the Invention I

Silica compositions prepared essentially as described in Comparative Example 1, Comparative Example 2, and/or as described in Comparative Example 3 above (20 g) were functionalised using trimethylchlorosilane reagent (TMS, 99.5%' Merck) (6 g), in toluene (90 mL) with imidazole (10 g) in a round bottomed flask (250 mL).

The solution was stirred and heated to 120°C for 17 hours. The mixture was cooled to 40°C EtOH (50 mL) was added before filtering through glass filter. The filtrate was washed in toluene (40 mL). The particles were refluxed in toluene:EtOH 50: 50 (150 mL) for one hour, cooled, filtered and washed with toluene and then ethanol.

The particles were then dried in a solvent oven at 90°C for 17 hours, resulting in a free flowing white powder.

Thalidomide was then encapsulated into the functionalised porous silica particles by a solvent impregnation and evaporation method, as described in Comparative Example 5 above. The physical properties of thalidomide-loaded silica particles produced according to the present example were even better than those described in Comparative Example 6.

Example 2

Dissolution Kinetics of Thalidomide Loaded Silica Particles

Dissolution kinetics of thalidomide-loaded silica particles produced according to Comparative Example 5 was measured in SLF (pH 7.4; made up with the salts NaCI, NaHCCb, KCI, MgCh, CaCh, NazSCU, sodium citrate dihydrate, NaHzPCU (all from Sigma)) at 37°C using a USP2 dissolution apparatus with stirring speed 50 rpm, with free, unloaded thalidomide being used as control. Concentration of drug at set times after release was measured by a UV/vis spectrometer (Cecil 3021) at 220 nm.

As shown in Figure 1, compared to the free drug control, dissolution kinetics of thalidomide was dramatically enhanced for all thalidomide-loaded samples.

When the same experiment was repeated for thalidomide-loaded silica particles produced according Example 1, dissolution kinetics are slower than for those samples prepared by Comparative Example 5.

Example 3 Animal Experiment

This study was intended to provide a pharmacokinetic assessment of inhaled thalidomide-loaded functionalised and non-functionalised silica particles produced according Example 1.

Twelve (for experiments with non-functionalised particles) + twelve (for experiments with functionalised particles) female rats (RccHa WIST, Envigo, Venray, the Netherlands), approximately 10 weeks old and weighing approximately 170-220 g were housed and acclimatised for at least 14 days before the start of dosing, during which period they were handled and trained in procedures they were later exposed to. Animals were randomized and housed 5/cage at 20-26°C and 40-70% RH. Drinking water and 2016 Teklad global 16% protein rodent diet, supplied by Envigo was freely available. Datesand F1850-Mouse Diet, High Fat, was given as treatment during the training and the dosing period, the total amount was <1% of the daily food consumption.

Premedication of the rats with 0.05 mg/mL of atropine s.c. (approximately 0. 1 mL/100 g body weight) was followed by an i.v. injection (in the tail vein) comprising a cocktail of 50 pg/mL of fentanyl and 1.0 mg/mL of medetomidine (Domitor Vet.) (50: 50), given gradually until anesthesia is achieved. The rats were intubated using a laryngoscope, with an appropriately-sized metal cannula inserted into the trachea.

Following anesthesia and tracheal intubation, the metal tracheal cannula was connected to Preciselnhale inhalation equipment (Inhalation Sciences Sweden AB). Animals were placed in the supine position during the inhalation procedure.

Thalidomide-loaded powder samples (obtained as described in Example 2 above) were loaded into the Preciselnhale dosing chamber in accurately- weighed appropriate doses. The actual inhaled doses of thalidomide were 0.097 mg/kg for the functionalised particles and 0.104 mg/kg for the non- functionalised particles. Sampling of blood and lungs (described below) was done immediately after and 0.5, 2 and 5 hours after dosing. There were three animals per dose group.

The sample was then ejected into a cylindrical holding chamber. From here, the aerosol was transferred by a constant airflow to the animal by a vacuum pump. The aerosol was then inhaled by the spontaneously breathing animal. The Preciselnhale system measured the particle concentration in the aerosol and the inhaled breathing volume, from these the inhaled dose could be calculated.

Prior to the study day, the Preciselnhale system was adjusted to the specific characteristics of the test formulation. Breathing tests were performed on stock animals of the same strain, sex, weight range and age as the study animals, and these data were be used to calculate the settings. Following the administration of the thalidomide loaded particles, animals were disconnected from the Preciselnhale equipment, extubated, and given an antidote of 0.05 mL of naloxone (0.4 mg/mL) and 0.05 mL Antisedan Vet. (5 mg/mL) to wake them up.

Monitoring was performed at specific time points after dosing, subsequently followed by euthanization and termination immediately after dosing and up to 5 hours after dosing as described above.

Animals were closely monitored for respiratory distress, changes in blood gases (pH, pCC>2, pC>2, and HCO3). A check of animal health and welfare status was done at least once on working days/weekend days/national holidays. A thorough examination for more permanent signs was performed at least once during the study.

In case of clinical symptoms in connection with dosing, animals were checked occasionally post-exposure and approximate time of recovery noted.

Blood was sampled (at least 0.6 mL in anticoagulant (EDTA)) at termination. Blood samples were centrifuged immediately after collection (approx. 2000xG, 5 min., 4°C) and plasma was transferred into separate polypropylene tubes and immediately frozen upright at approximately -70°C within 5 min after centrifugation. Animals were euthanized by exsanguination from a common carotid artery under isoflurane and oxygen anesthesia, and whole lungs were excised at the time points described above. Tracheas were cut at the same distance from the distal end of the larynx in all animals to secure that the anatomic region around the distal end of the tracheal cannula was included. Excised tracheas and lungs were snap frozen in one piece in liquid N2, then transferred to -70°C. The excised tracheas and lungs were weighed for wet weight.

The extraction of thalidomide from plasma was performed by precipitation of proteins using acetonitrile with 0.1% formic acid (FA) and internal standard (thalidomide-d4). Lung samples were homogenized with 5 ml acetonitrile with 0.1% FA after weighing. The plasma and homogenized lung samples were the centrifuged for 10 minutes (4000 RPM) at room temperature. The supernatant was transferred and analysed. All samples were analysed by first separating them by reversed phase gradient HPLC and subsequently detecting them using positive electrospray ionization and multiple reaction monitoring. Quantification was performed in the range 1 ng/mL to 3000 ng/mL of thalidomide.

Lung and plasma concentration-time data profiles for thalidomide were evaluated by a pharmacokinetics expert performing non-compartmental analysis (sparse sampling) using Phoenix 64 WinNonlin® from Certara (Princeton, NJ, USA). All samples demonstrated measurable levels of thalidomide.

The mean Cmax was 89 ng/mL in plasma and 671 ng/g lung tissue in animals administered non-functionalised silica particles, while the corresponding values for Cmax after administration of functionalised particles was 266 ng/mL in plasma and 4310 ng/g lung tissue.

The mean AUCiast was 219 h*ng/mL in plasma and 244 h*ng/g in lung in animals administered non-functionalised particles, while the corresponding values for AUCiast after administration of functionalised particles was 445 h*ng/mL in plasma and 1810 h*ng/g in lung.

The lung exposure of thalidomide was thus much higher after administration of the functionalised silica particles as compared to non-functionalised particles.

Example 4

Manufacture of a Composition of the Invention II

Silica compositions were functionalised using triethylchlorosilane reagent (TES; Sigma Aldrich) essentially as described in Example 1 above.

Thalidomide was then encapsulated into the functionalised porous silica particles by a solvent impregnation and evaporation method, as described in Comparative Example 5 above. Example 5

Manufacture of a Composition of the Invention III

5 to 10 g of a silica composition prepared essentially as described in Comparative Example 1, Comparative Example 2, and/or as described in Comparative Example 3 above is dispersed in toluene (50 mL).

Ethylchloroformate (1 g) is added in presence of N-methylmorpholine (0.5g) followed by stirring at 80°C for 18 hours.

The product is filtered, washed with tetrahydrofuran and dried at 90°C. The samples are analysed by TGA, carbon load and IR.

Thalidomide is then encapsulated into the functionalised porous silica particles as described in Comparative Example 5 above.

Example 6

Manufacture of a Composition of the Invention IV

5 to 10 g of a silica composition prepared essentially as described in Comparative Example 1, Comparative Example 2, and/or as described in Comparative Example 3 above is dispersed in tetrahydrofuran (50 mL).

Ethyl iodide (1 g) and sodium carbonate (0.2 g) are added and refluxed for 24 hours.

The product is filtered, washed with EtOH:water to remove any salts and dried at 90°C. The samples are analysed by TGA, carbon load and IR.

Thalidomide is then encapsulated into the functionalised porous silica particles as described in Comparative Example 5 above.