Improved nanoseeding process

Abstract

What is described herein relates to a method for generating micro-particles comprising the steps of providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm and 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 1 and 1000 optionally isolating micro-particles.

Description

[0001] BHC243022-FC

[0002] -1-

[0003] IMPROVED NANOSEEDING PROCESS

[0004] The present invention relates to a nano-seeding process.

[0005] BACKGROUND

[0006] A significant proportion of active ingredients e.g. of active pharmaceutical ingredients and active agrochemical ingredients have poor solubility and / or require a high bioavailability and short dissolution times. To improve the poor solubility especially in water and optimize high bioavailability and short dissolution times, one method is to reduce the particle size of the active ingredients to create a large surface-to-volume ratio and hence a high solubility particle interface.

[0007] A well-known method to produce microparticles is dry-milling. However, this method has some limitations, such as high energy consumption, low yields, formation of encrustations, and difficult- to-control particle sizes and surface properties, which complicate microparticle production and consistent quality. Therefore, there is a need for an improved process for generating micro-particles. This need is met by a simplified process for generating micro-particles.

[0008] What is described herein relates in a first aspect to a method for generating micro-particles comprising the steps of a) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of the active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b)providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the same active ingredient in the range of 2 to 20, d) optionally isolating micro-particles. BHC243022-FC

[0009] -2-

[0010] DESCRIPTION OF THE DRAWINGS

[0011] The various aspects of the subject-matter described in this application are illustrated by the drawings which are not meant to be limiting in any way. Instead the drawings merely serve to illustrate the subject-matter described herein. Hence, the embodiments depicted in the drawings are of exemplary nature only.

[0012] Fig.l depicts a comparison between the particle size distribution resulting from a method for generating micro-particles as described in EP4154872 (Example la / grey line) and the method described herein (cf. Example Ib / black dotted line). Even though the aqueous nanosuspension used in the method described herein (black dotted line) only comprised 0.198 wt% seeding material whereas the aqueous nanosuspension used in the method for generating micro-particles as described in EP4154872 comprised 1.9 wt% seeding material (where the % wt is calculated with respect to the total concentration of substrate i.e. a substance here an active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined) the method described herein resulted in microparticles of the same or a smaller size than microparticles generated with the method as described in EP4154872. As detailed below the particle size and its distribution can be measured by the methods of Dynamic Light Scattering (DLS), laser diffraction or electron microscopy as described below. Hence the method described herein is advantageous as it requires less nanoparticle material.

[0013] Fig. 2 depicts an exemplary set-up for carrying on the methods described herein. All connections shown between different parts can for example be realized using tubes made from polytetrafluoroethylene (PTFE). The set up comprises a first storage vessel (1) and a second storage vessel (2). In this Example the first storage vessel (1) contains the aqueous nanosuspension and the second storage vessel (2) contains the homogenous saturated solution. The storage vessels (1) and (2) are connected to the mixing element (3a). The mixing element (3a) is connected to a collecting vessel (4). The aqueous nanosuspension and the homogenous saturated solution are transported via the action of pumps (5) and (6) respectively to the mixing element (3a). Via adjusting the speed of pumps (5) and (6) as well as the dosing valves (7) and (8) the flow rates of the aqueous nanosuspension and the homogenous saturated solution can be altered to influence the degree of generated supersaturation that initiates the particle growth and thereby the production of microparticles. The microparticles containing suspension is collected in the collecting vessel (4). The mixing element can comprise a discharge channel, in which the ripening processes of the microparticles can continue before the microparticles containing suspension is collected in the collecting vessel. Finally, the growth of the microparticles can optionally be quenched with pure BHC243022-FC

[0014] -3- antisolvent (aqueous solution) in the collecting vessel (4). Optionally the microparticles can be isolated via filtering after leaving the discharge channel of the mixing element (3 a).

[0015] Fig. 3 depicts the same exemplary set-up as Fig. 2 with the addition of another mixing element (3b) that is connected to the second storage vessel (2) that contains the homogenous saturated solution, a third pump (9) and a third dosing valve (10). Again all connections shown between different parts can for example be realized using tubes made from polytetrafluoroethylene (PTFE). The operation differs from the operation of the set-up depicted in Fig. 2 in that after the microparticle containing suspension leaves the mixing element it is not immediately collected in the collection vessel (4), but further homogenous saturated solution is added to the microparticle containing suspension in the second mixing element (3b). The amount of homogenous saturated solution added in mixing element (3b) can be adjusted via the dosing valve (10) and / or via the speed of pump (9). Hence the microparticles in the microparticle containing suspension coming from the first mixing element (3 a) act as seeds for further growth of the particles until the desired particle size is reached in the mixing element (3b) (or its discharge channel). After leaving the discharge channel of the second mixing element (3b) the growth of the microparticles can optionally be quenched with pure antisolvent (aqueous solution) in the collecting vessel (4). Optionally the microparticles can be isolated by filtering after leaving the discharge channel of the mixing element (3b).

[0016] Fig. 4 depicts another exemplary set-up. Again all connections shown between different parts can for example be realized using tubes made from polytetrafluoroethylene (PTFE). In comparison to Fig. 3 this Example set up also comprises a third mixing element (3c), another pump (11) and another dosing valve (12) as well as optional dosing valve (13). Moreover, here storage vessel (1) only comprises a solution of water and at least one stabilizer. In operation the aqueous solution is transported from storage vessel (1) by action of pump (5) and via dosing valve (7) to a first mixing element (3a). Simultaneously a homogenous saturated solution of the substance in a crystallization medium is also transported from storage vessel (2) by action of pump (6) and via dosing valve (8) to the first mixing element (3a). In the mixing element (3a) the supersaturation level is sufficient to initiate nucleation and thereby nanoparticles are formed. Subsequently the suspension comprising nanoparticles enters mixing element (3b) where it is mixed with more homogenous saturated solution of the substance in a crystallization medium from storage vessel (2) by action of pump (9) and via dosing valve (10). Thus the nanoparticles generated in mixing element (3a) act as seeding material (i.e. as “nanoseeds”) in mixing element (3b). The generated microsuspension then enters a third mixing element (3c) in which it is mixed with additional aqueous solution comprising at least one stabilizer from storage vessel (1) by action of pump (11) and via dosing valve (12) in order to quench and stabilize the microsuspension which is then collected in the collecting vessel (4). BHC243022-FC

[0017] -4-

[0018] Optionally the in situ prepared nanosuspension is first quenched with aqueous solution comprising at least one stabilizer from storage vessel (1) via dosing valve (13) before it enters the second mixing element (3b) to stabilize the nanosuspension if necessary. This option is depicted as dotted line. Fig. 5 shows a photograph of a T mixer as it was used in Example 3.

[0019] Fig. 6 depicts a comparison between the particle size distribution resulting from the method for generating micro-particles as described herein in Example 1c. The aqueous nanosuspension of the black dotted line comprised 0.198 wt% seeding material whereas the aqueous nanosuspension of the solid black line comprised only IO'8wt% seeding material (where the % wt is calculated with respect to the total concentration of substrate i.e. a substance here an active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined). The result demonstrated that the particles were larger if almost no seeding material was present and hence confirmed the modelled value VIII of Table 2. This was the case even though Example 1c did not employ nanoparticles which were characterized by a size of a d90 in the range of 10 nm and 999 nm as measured by Dynamic Light Scattering (DLS) in at least one of length, diameter or height as well as a size of a d90 in the range of 1pm and 2pm as measured by Dynamic Light Scattering (DLS) in at least one of length, diameter or height as measured by Dynamic Light Scattering (DLS) but used nano-particles that were characterized by a nano-particle size in all of length, diameter or height of a d90 in the range of 10 nm and 999 nm.

[0020] Fig.7 depicts a comparison between the particle size distribution resulting from a method for generating micro-particles as described in EP4154872 (grey line) and the method described herein (cf. Example Id / black dotted line) for a different substance i.e. a different active ingredient than shown in Fig. 1. Even though the aqueous nanosuspension used in the method described herein (black dotted line) comprised 2 wt% seeding material and also the aqueous nanosuspension used in the method for generating micro-particles as described in EP4154872 comprised 2 wt% seeding material (where the % wt is calculated with respect to the total concentration of substrate i.e. a substance here an active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined) the method described herein resulted in microparticles of a smaller size than microparticles generated with the method as described in EP4154872. In other words, less wt% seeding material could have been used in the method described herein to still generate microparticles of the same size with the method as described in EP415487. How much seeding material could have been used can be calculated using the method described in Example 3a.

[0021] Fig. 8 depicts the temperature profile of Example 2. BHC243022-FC

[0022] -5-

[0023] DETAILED DESCRIPTION

[0024] It was surprisingly found that, using the method described herein, less nanoparticles as seeding material were required in the aqueous nanosuspension comprising nanoparticles as seeding material compared to the method as described in EP4154872 while still generating microparticles of a similar size (using the d50 or d90 value calculated from a particle size distribution) as those generated with the method as described in EP4154872 (cf Fig. 1). This was the case even though in other settings the high supersaturation levels used in the method described herein can cause uncontrolled, spontaneous primary nucleation and subsequent growth - which is to be avoided since the resulting particles vary greatly in size and it bears the risk that other or more than one polymorph is formed (Ouyang et al, 2021, CrystEngComm, 2021, 23, 813-823).

[0025] Accordingly what is described herein relates in a first aspect relates to a method for generating micro-particles comprising the steps of a) Providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 1 to 1000, d) optionally isolating micro-particles.

[0026] As in the method described herein in the first aspect the provided aqueous nanosuspension comprising nanoparticles as seeding material was mixed with the provided homogenous saturated solution the nanosuspension acted as an antisolvent for the substance e.g. the active ingredient dissolved in the homogenous saturated solution resulting in a rapid distribution of the nanoparticles of the aqueous nanosuspension throughout the entire mixed volume in a very short time. In turn this lead to a very rapid supersaturation or increase of supersaturation followed by precipitation of the substance e.g. the active ingredient. Consequently, due to the high supersaturation level an aqueous nanosuspension comprising a lesser amount of nanoparticles was needed compared to the method as described in EP4154872. In addition, the method described herein is better suited for large scale BHC243022-FC

[0027] -6- industrial application. Moreover, the combination of controlled but very rapidly developing supersaturation or increase of supersaturation avoided occurrence of primary nucleation. Hence, as demonstrated in Example Id this avoidance of primary nucleation also avoided the occurrence of a polymorph with a different crystal structure than the nanoparticles acting as seeding material (“nanoseeds”) of the aqueous nanosuspension. Thus, the method described herein in the first aspect in addition to the above advantages is favourable for a substance generating at least two polymorphs where the undesirable polymorph exhibits the faster nucleation in cooling and / or antisolvent crystallization.

[0028] Anti-solvent crystallisation is a technique that utilizes the addition of a so-called antisolvent to reduce the solubility of the target compound in a solution constituted by a solvent where the solubility is higher, i.e. the chemical potential of the target compound in the mixed solvent / antisolvent is higher than in the solvent alone (cf. for example Beckmann Chapter 9.1 Generation of Supersaturation in a Crystallizer Crystallization: Basic Concepts and Industrial Applications 2013)

[0029] To a person skilled in the art, it is clear that as in any process based on anti-solvent crystallisation - also termed salting out - also in the method described herein the crystallization medium in which the substance is provided in is miscible with the anti-solvent. Hence in the method for generating micro-particles described herein the crystallization medium is a solvent that is miscible with the aqueous nanosuspension.

[0030] As used herein the term “substance” refers to any material that crystallizes. It is clear to a person skilled in the art that any substance can be used in the method described herein, as long as it can crystallize.

[0031] In a preferred embodiment the substance is an active ingredient i.e. an agent, compound, substance, compositions, or mixtures thereof, that provides a pharmacological and / or agrochemical effect.

[0032] In an even more preferred embodiment, the active ingredient is an agent, compound, compositions, or mixtures thereof, that provides a pharmacological and / or agrochemical effect and which has a melting temperature above 30°C (i.e. above room temperature). This is advantageous as it allows for a more economic process since the reaction does not have to be cooled.

[0033] In a highly preferred embodiment, the active ingredient is an agent, compound, compositions, or mixtures thereof, that provides a pharmacological and / or agrochemical effect and which has a melting temperature above 30°C (i.e. above room temperature) and has a molecular mass in the range of 100 BHC243022-FC

[0034] -7-

[0035] Da and 2000 Da, preferably in the range of 200 da to 1500 Da even more preferably in the range of 300 Da to 1000 Da and a melting temperature above 30°C (i.e. above room temperature).

[0036] In a further embodiment, the active ingredient is selected from the list consisting of abarelix, abemaciclib, abiraterone, acalabrutinib, aclarubicin, acetyl salicylic acid (aspirin), afatinib, aflibercept, aldesleukin, alectinib, alendronic acid, alfaferone, alitretinoin, almonertinib, alpelisib, alpharadin, altretamine, amifostine, aminoglutethimide, aminolevulinic acid, amrubicin, amsacrine, anastrozole, ancestim, anethole dithiolethione, angiotensin II, antithrombin III, apalutamide, aprepitant, arglabin, avapritinib, axicabtagene ciloleucel, axitinib, azacitidine, beclomethasone dipropionate, belinostat, belotecan, bendamustine, bexarotene, bicalutamide, binimetinib, bisantrene, bleomycin, boanmycin hydrochloride, borofalan, bortezomib, bosutinib, budesonide, buserelin, cabazitaxel, cabozantinib, calcitonine, calcium folinate, calcium levofolinate, capecitabine, capmatinib, carbamazepine, carboplatin, carboquone, carfilzomib, carmofur, carmustine, catequentinib, celecoxib, celmoleukin, ceritinib, chlorambucil, chlormadinone, chlormethine, chloroxoquinoline, cidofovir, cinacalcet, cisplatin, cladribine, clodronic acid, clofarabine, cobimetinib, copanlisib, crisantaspase, crizotinib, cyclophosphamide, cyproterone, cytarabine, dabrafenib, dacarbazine, dacomitinib, dactinomycin, darbepoetin alfa, dasatinib, daunorubicin, decitabine, deferasirox, degarelix, denileukin diftitox, depreotide, deslorelin, dexrazoxane, dianhydrogalactitol, docetaxel, dolasetron, doxifluridine, doxorubicin, dronabinol, duvelisib, elliptinium acetate, eltrombopag, enasidenib, encorafenib, endostatin, enocitabine, enzalutamide, epirubicin, epitiostanol, epoetin alfa, epoetin beta, epoetin zeta, eptaplatin, eribulin, erlotinib, ensartinib, entrectinib, erdafitinib, esomeprazole, estradiol, estramustine, estrone, ethinylestradiol, etoposide, everolimus, evocalcet, exemestane, fadrozole, famotidine, fentanyl, filgrastim, flumatinib, fluoxymesterone, fluticasone, fluticasone furoate, floxuridine, fludarabine, fluorouracil, flutamide, folinate, folinic acid, formestane, forodesine, fosaprepitant, fotemustine, fruquintinib, fulvestrant, gadobutrol, gadoteridol, gadoteric acid meglumine, gadoversetamide, gadoxetic acid, gallium nitrate, ganirelix, gefitinib, gemcitabine, gendicine, gilteritinib, glasdegib, glucarpidase, glutoxim, goserelin, granisetron, granulocyte colony stimulating factor, hematoporphyrin, histamine dihydrochloride, histrelin, hydroxycarbamide, ibandronic acid, idarubicin, idelalisib, ifosfamide, imatinib, imiquimod, improsulfan, immunocyanin, indisetron, incadronic acid, ingenol mebutate, interferon alfa, interferon beta, interferon gamma, lanreotide, lansoprazole, lapatinib, lenalidomide, lenograstim, lentinan, lenvatinib, letrozole, leuprorelin, levamisole, levonorgestrel, levothyroxine sodium, lisuride, lobaplatin, lomustine, lonidamine, lorlatinib, lurbinectedin, luspatercept, mafodotin, masoprocol, medroxyprogesterone, megestrol, melarsoprol, melphalan, mepitiostane, mercaptamine, mercaptopurine, mesna, methadone, methotrexate, methoxsalen, methylaminolevulinate, methylprednisolone, methyltestosterone, metirosine, midostaurin, mifamurtide, mifepristone, miltefosine, miriplatin, mitobronitol, BHC243022-FC

[0037] -8- mitoguazone, mitolactol, mitomycin, mitotane, mitoxantrone, molgramostim, mometasone, monosodium alpha luminol, mopidamol, morphine hydrochloride, morphine sulfate, nabilone, nabiximols, nafarelin, naloxone, naltrexone, nartograstim, nedaplatin, nelarabine, neratinib, neridronic acid, netupitant / palonosetron, nilotinib, nilutamide, nimorazole, nimustine, nintedanib, niraparib, nitracrine, octreotide, olaparib, olmutinib, omacetaxine mepesuccinate, omeprazole, ondansetron, oprelvekin, orelabrutinib, orgotein, orilotimod, osimertinib, oxaliplatin, oxycodone, oxymetholone, ozogamicine, paclitaxel, padeliporfm, palbociclib, palifermin, palonosetron, pamidronic acid, panobinostat, pantoprazole, pazopanib, pegaspargase, pemigatinib, pegfilgrastim, peginterferon alfa-2b, pemetrexed, pentazocine, pentostatin, peplomycin, perflubutane, perfosfamide, phenoxybenzamine, picibanil, pilocarpine, pirarubicin, pixantrone, plerixafor, plicamycin, poliglusam, pomalidomide, ponatinib, porfimer sodium, pralatrexate, pralsetinib, prednimustine, prednisone, procarbazine, procodazole, propranolol, quinagolide, quizartinib, rabeprazole, radotinib, raloxifene, raltitrexed, ramosetron, ranimustine, rasburicase, razoxane, refametinib, regorafenib, relugolix, ribociclib, ripretinib, risedronic acid, rivoceranib, rolapitant, romidepsin, romiplostim, romurtide, rucaparib, govitecan, lexidronam, sargramostim, secretin, selinexor, selpercatinib, selumetinib, sipuleucel-T, sirolimus, sizofiran, sobuzoxane, sodium glycididazole, sonidegib, sophoridine hydrochloride, sorafenib, stanozolol, streptozocin, sunitinib, surufatinib, tagraxofusp, talaporfm, talazoparib, talimogene laherparepvec, tamibarotene, tamoxifen, tapentadol, tasonermin, tazemetostat, teceleukin, merpentan, tegafur, temozolomide, temsirolimus, teniposide, testosterone, tetrofosmin, thalidomide, thiotepa, thrombopoietin, thymalfasin, thyrotropin alfa, tioguanine, tirabrutinib, tisagenlecleucel, tivozanib, topotecan, toremifene, trabectedin, trametinib, tramadol, treosulfan, tretinoin, trifluridine + tipiracil, trilostane, triptorelin, trofosfamide, tryptophan, tucatinib, tucidinostat, ubenimex, ulipristal, umbralisib, valatinib, valrubicin, vandetanib, vapreotide, vemurafenib, venetoclax, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, vismodegib, vorinostat, vorozole, zanubrutinib, zinostatin, zinostatin stimalamer, zoledronic acid, zorubicin.

[0038] In a most preferred embodiment the active ingredient is selected from the group consisting of Darolutamide, Finerenone, Vericiguat, Asundexian, Aflibercept, Nurandociguat, (R)-2-(N-[4- amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide.

[0039] In one embodiment the active ingredient is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]- 4-fluoro-anilino)propanamide.

[0040] As used herein, the term “particle” refers to a solid, gel, or semisolid material having a relatively small size. BHC243022-FC

[0041] -9-

[0042] As used herein the term “micro-particle” which is used synonymously with microparticle refers to particles having a d90 particle size in the range of > 999 nm to 100pm in length, width and height.

[0043] As used herein the term “nano-particle” which is used synonymously with nanoparticle refers to particles having a d90 particle size in the range of > 10 nm to < 999 nm in at least one of length, or width, or height.

[0044] The terms d50 or d90 mean that 50% or 90% of the total particle volume of a given sample consists of particles smaller in length, or width, or height than the given value for d50 or d90. The particle size is determined by methods known in the art such as laser diffraction, electron microscopy or Dynamic Light Scattering (DLS).

[0045] In a preferred embodiment the particle size is determined by Dynamic Light Scattering (DLS). Here and further on, it is a volume based particle size determination if it is not stated otherwise and the resulting d50 or d90 values are a volume based d50 or a volume based d90 respectively.

[0046] The particle size distribution as well as the d50 and d90 particle sizes can be calculated from the measured laser diffraction or DLS results as published by the American Society for Testing and Materials (ASTM International) under the Designation E799 - 03. Herein the term d50 or d90 are synonymous to Dvo.s or Dvo.y respectively. In this respect it is clear to a skilled person that the d50 value does not necessarily have to correspond exactly to the mathematical average. This is only the case in special situations, for example when the size distribution is symmetrical. The d50 value rather corresponds to the median of the distribution.

[0047] A person skilled in the art is capable of choosing technical equipment suitable for achieving a respective d50 or d90 value. For example, it is clear to a skilled person that using an agitated ball mill will result in a smaller d50 and d90 values than using a planetary ball mill, especially if the agitated ball mill is cooled.

[0048] If laser diffraction was used to determine the particle size said laser diffraction was performed in a Malvern Mastersizer 3000. A spatula tip of the powder sample was suspended in 10 ml of deionized water and a droplet of Tween80 was added as dispersant. The sample was first mixed with the spatula then dispersed with a pipette. The Mastersizer 3000 was equipped with a Hydro MV unit, where deionized water was provided to disperse and dilute the sample further. The dispersed sample was added to the Hydro MV unit until the obscuration reached 5%. The mixer of the Hydro MV BHC243022-FC

[0049] -10- unit was set to 2500 rpm. The first three measurements were performed with red and blue laser lights using the Mie light scattering model. Three measurements were performed. In addition, ultrasonic was applied for 1, 3 and 5 min. After each Bonification time one measurement was performed. The particle size typically did not change after 3 minutes of Bonification. Therefore, this value was taken for the comparison of experiments.

[0050] X-Ray diffraction (XRD) on the other hand can be used to determine the atomic and molecular structure of a crystal. XRD is based on the fact that a crystalline structure causes a beam of incident X-rays to diffract into many specific directions. By measuring the angles and intensities of these diffracted beams, a crystallographer can produce a three-dimensional picture of the density of electrons within the crystal. From this electron density, the mean positions of the atoms in the crystal can be determined, as well as their chemical bonds, their crystallographic disorder, and various other information, (see, for example, Dann, Sandra E. Reactions and characterization of solids. Vol. 2. Royal Society of Chemistry, 2000. or Chauhan and Chauhan, J Anal Bioanal Tech 2014, 5:5).

[0051] As used herein the term “seeding material” also termed nanoseeds refers to nano-particles of the same substance or composition as the micro-particles to be generated by the method described herein in the first aspect. In general the seeding material is analyzed for its quality, purity, and particle size distribution. The quality of the seeding material i.e. the polymorphism and / or pseudopolymorphism can be determined, e.g. by means of X-ray diffraction and / or Raman and / or IR spectroscopy. The size distribution of the seeding material can be determined by laser diffraction and / or dynamic light scattering. The purity of the seeding material can be determined e.g. by chromatography.

[0052] In case the seeding material has a strong anisotropic asymmetry among its characteristic axes of growth, it is of needle-like and platelet-like appearance. These particles are termed “asymmetric nano-particles” herein, if at least one of length or width or height is characterized by i) a size of a d90 e.g. a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS) as well as ii) a size of a d90 e.g. a volume based d90 in the range of 1pm to 2pm as measured by Dynamic Light Scattering (DLS)

[0053] The characterization of the smallest dimension can be performed either via laser diffraction or via electron microscopy (SEM or TEM). Sizing techniques (membrane filtration) can also be utilized to verify if all characteristic dimensions of a multidimensional particle are below a certain threshold. BHC243022-FC

[0054] -11-

[0055] As used herein the term “homogenous saturated solution” refers to a solution in which the concentration of a solute equals the maximum amount of that solid that can be dissolved in that particular solution at a given temperature and pressure.

[0056] As used herein the term “supersaturated” refers to a solution or a suspension, in which the concentration of a solute exceeds the maximum amount of that solid that can be dissolved in that particular solution at a given temperature and pressure for any solute with a size greater than > 999 nm in length, width and height.

[0057] As used herein the term “undersaturated” refers to a solution or a suspension, in which the concentration of a solute is below the maximum amount of that solid that can be dissolved in that particular solution at a given temperature and pressure for any solute with a size greater than > 999 nm in length, width and height.

[0058] A person skilled in the art knows how to determine the maximum amount of a solid that can be dissolved in a particular solution (cf. for example Beckmann 2013, pages 53 and 54).

[0059] To a person skilled in the art, it is clear that the concentration of the substrate i.e. a substance e.g. an active ingredient during and possibility after mixing of the homogenous saturated solution with the aqueous nanosuspension comprising seeding material exceeds the concentration of the substrates (substances) e.g. an active ingredients thermodynamically equilibrium value.

[0060] A person skilled in the art knows how to provide a supersaturated solution or a supersaturated suspension and which equipment (cf. e.g. Beckmann 2013 Chapter 9.1 Generation of Supersaturation in a Crystallizer Crystallization: Basic Concepts and Industrial Applications).

[0061] If a supersaturated homogenous solution is used in the method described herein in the first aspect it is in the metastable zone width. Metastable zone width as used herein refers to the interval of temperature and concentration in which a solution is supersaturated, but kinetically stable, i.e. the spontaneous appearance and growth of crystals whilst thermodynamically possible, is not observed or is strongly delayed. In other words, a skilled person is aware of the fact that the metastable zone width refers to the combination of temperature range and concentration range i.e. the zone in which a solution of a given solute is supersaturated but does not yet spontaneously crystallize.

[0062] The metastable zone width i.e. the temperature and concentration values for a given solute in a given crystallization medium have to be determined on a case by case basis. A person skilled in the art knows how to perform the required experiments for a given solute in a given crystallization medium. For example, a skilled person would perform a screening of solvents compatibly with BHC243022-FC

[0063] -12- chemical constraints such as product degradation followed by screening for the possible presence of (pseudo)polymorphs and measuring the relevant solubilities. Alternatively, computer models can also be exploited to a certain degree to mitigate (but not fully replace) in vitro experiments, e.g. by performing DFT or MD simulations to assess the relative stability of different polymorphs, or by estimating the solubility, e.g. using PC-SAFT and the related theory of groups.

[0064] Homogenous as used herein refers to the fact that a material is uniform in appearance. When referring to a solution homogenous therefore means that no distinct solid and liquid phases co-exist.

[0065] A skilled person is aware of the fact that upon spontaneous formation of particles, or after their addition through seeding, the supersaturated solution becomes a suspension, i.e. a system where distinct solid and liquid phases co-exist.

[0066] When referring to a suspension - e.g. after the homogenous saturated solution and the seeding material have been brought into contact - “homogenous” is herein meant to be if no spontaneous segregation of particles according to their size can be detected within the suspension and / or if no dead volumes can be observed, e.g. by means of visual analysis or tracer-based methods, or predicted, e.g. by means of CFD simulations within 30 min to 2 days after preparation of the solution. The selection of the relevant operating conditions to ensure good miscibility can be assessed by calculating the relevant non-dimensional numbers (e.g. Newton, Reynold, Peclet), which account for the geometric properties of the chosen vessel and impeller. The correlations between such numbers and the relevant operating conditions are tabulated in the relevant literature, e.g. Perry Chemical Engineering Handbook, 8th Edition, McGraw-Hill, 2007.

[0067] As used herein the term “stabilizer” refers to a polymer that sterically and / or a surfactant that electrostatically or sterically stabilizes the nano-particles.

[0068] As used herein the term “supersaturation level” refers to the maximum supersaturation level that is attainable by the system, if neither nucleation nor growth were active, thereby consuming the solute from the solution. The calculation of the maximum supersaturation level as used herein is performed by dividing the

[0069] • overall concentration of substance e.g. an active ingredient upon mixing in the homogenous solution and the aqueous nanosuspension combined

[0070] • by the solubility of the substance e.g. the active ingredient in the crystallisation medium.

[0071] For example BHC243022-FC

[0072] -13-

[0073] • if the initial concentration of the substance e.g. an active ingredient in homogenous saturated solution and the aqueous nanosuspension combined is assumed to be 0.044 g / gsoiution and

[0074] • if the solubility of the substance e.g. that active ingredient is assumed to be 0.01 g / gsoiution

[0075] Then the supersaturation level - also referred to as supersaturation ratio - would be 0.044 g / gsoiution divided by 0.01 g / gsoiution equaling 4,4 (without a unit).

[0076] It should be noted that the solubility of the substance e.g. an active ingredient in the crystallisation medium refers to the solubility at (thermo)dynamic equilibrium - also called “bulk solubility” - of the substance e.g. the active ingredient in the crystallisation medium, i.e. it is assumed that all particles of the substance e.g. the active ingredient forming the solid phase in equilibrium with the liquid phase behave if they were of infinite size (cf for example Beckmann 2013, pages 53 and 54).

[0077] As used herein the term “crystallization medium” refers to a liquid in which the substance e.g. an active ingredient constituting the nano-particles and the to be generated micro-particles is soluble in and from which the substance e.g. the active ingredient can crystallize when the medium is supersaturated. A skilled person is aware of the fact that in cases where more than one active ingredient is present in a given composition e.g. active ingredient combinations in step b) the substance e.g. the active ingredient to be crystallized is present as the homogenous saturated solution while possible other (active) ingredients in the crystallization medium are present as suspension.

[0078] A person skilled in the art knows how to determine a suitable crystallization medium depending on the specific process as well as chemical and regulatory requirements. Standard crystallization media for pharmaceutical and agro-chemical relevant substances e.g. active ingredients are, for example, water, methanol, ethanol, acetone, acetic acid, depending on whether an inorganic (water) or organic (other) medium is required, whether the solubility is promoted by protonated (water, alcohols, acids) or non-protonated (acetone or esters) solvents and / or by the presence of H-bonds and / or other coordination groups.

[0079] The nanoparticles e.g. the active ingredient nanoparticles of the aqueous nanosuspension are preferably generated via nano-grinding (also termed nano grinding, nanogrinding or nanomilling). Suitable nanogrinding methods can be selected from the group consisting of wet bead milling in stirred media mills i.e. agitated ball mills, wet bead milling in planetary mills - especially suitable for small amounts- high pressure homogenization. Alternatively, the application of high shear BHC243022-FC

[0080] -14- forces in aqueous suspensions like in a high pressure homogenization process or the application of high impact forces between the particles like in a microfluidizer can be used to produce nanoparticles as well as acoustic comminution technology (e.g. Dennis H. Leung et. Al, A new and improved method for the preparation of drug nanosuspension formulations using acoustic mixing technology, Int. J. Pharrn. 473(2014)10-19).

[0081] Correspondingly also the aqueous nanosuspension comprising nanoparticles e.g. active ingredient nanoparticles as seeding material is preferably generated via nano-grinding using wet bead milling in stirred media mills. An example for preparing the aqueous nanosuspension comprising nanoparticles e.g. active ingredient nanoparticles as seeding material is given in the description of Example 1. However, the skilled person readily understands that the amount of substance microcrystals, the amount of stabilizer(s), the exact mill used as well as the milling duration and temperature depend on active ingredient employed and the total amount of aqueous nanosuspension comprising nanoparticles as seeding material prepared.

[0082] To a skilled person it is clear that if nano grinding is used for providing the seeding material said nano grinding can only be performed if the nano -particles e.g. active ingredient nanoparticles are not completely dissolved. Hence the skilled person would choose a suitable temperature within a temperature range of 253 K to 323 K, preferably in the range of 273 K to 303 K for nano-grinding the nano-particles in the aqueous suspension.

[0083] Preferably the micro-particles obtained with the method for generating micro-particles described herein have a d90 particle size in the range of 2pm to 50pm, more preferably b in the range of 2pm to 30pm, most preferably between 2pm to 20pm.

[0084] It is preferred that the nanoparticles e.g. active ingredient nanoparticles have a d90 e.g. a volume based d90 particle size in the range of > 10 nm to < 999 nm, preferably > 10 nm to < 700 nm preferably > 10 nm to < 400 nm especially preferred a d90 particle size in the range of > 10 nm to < 999 nm and most preferred an d90 particle size in the range of > 10 nm to < 200 nm in at least one of length, or width, or height.

[0085] Alternatively is preferred that the nanoparticles (also termed nano -particles) have a d50 e.g. a volume based d50 particle size in the range of > 10 nm to < 999 nm, preferably > 10 nm to < 700 nm , preferably > 10 nm to < 400 nm especially preferred a d50 particle size in the range of > 10 nm to < 999 nm and most preferred an d50 particle size in the range of > 10 nm to < 200 nm in at least one of length, or width, or height.

[0086] In one embodiment of the method for generating microparticles of the first aspect the crystallization medium and hence the solvent of the substance e.g. the active ingredient in the homogeneous BHC243022-FC

[0087] -15- saturated solution is selected from the group consisting of low molecular organic solvents with melting point <20°C at normal pressure (Mw < 200g / mol, e.g. acetic acid, lactic acid, formic acid, citric acid, oxalic acid, malic acid, tartaric acid, butyric acid, and there like), acetone, acetonitrile, 1,2-butanediol, 1,3 -butanediol 1,4-butanediol, 2 -butoxy ethanol, diethanolamine, diethylenetriamine, dimethoxyethane, , dimethyl sulfoxide (DMSO), dioxane, ethanol, ethylamine, ethylene glycol, glycerol, isopropanol, methanol, n-propanol, methyl diethanolamine, 1,3- propanediol, propanoic acid, propylene glycol, 1,5 -pentanediol, sulfolane, pyridine, tetrahydrofuran (THF), triethylene glycol.

[0088] These are all crystallization media / solvents miscible with the aqueous nanosuspension.

[0089] In a preferred embodiment the crystallization medium and hence the solvent of the substance e.g. the active ingredient in the homogenous saturated solution is selected from the group consisting of acetic acid, isopropanol, ethanol, methanol, acetone, DMSO, Dioxane, THF.

[0090] In the case of acetylsalicylic acid as active ingredient a mixture of acetic anhydride and acetic acid is used as crystallization medium, due to the requirements of synthesis occurring prior to crystallization. Another known crystallization medium is ethanol.

[0091] In one embodiment the method described herein in the first aspect is a method for generating microparticles comprising the steps of a) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer b) providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the same active ingredient in the range of 2 to 20, d) optionally isolating micro-particles.

[0092] As demonstrated in Fig. 7 this embodiment has the advantage that due to the higher supersaturation level smaller microparticles can be generated with the same amount of seeding material than if the method for generating micro-particles as described in EP4154872 is employed. BHC243022-FC

[0093] -16-

[0094] In one embodiment of the method for generating microparticles of the first aspect the aqueous nanosuspension comprising nanoparticles e.g. active ingredient nanoparticles as seeding material comprises a concentration in the range of 0.99 % wt to of 0.001% wt preferably in the range of 0.99 % wt to 0.001% wt more preferably in the range of 0.9 % wt to 0.7 % wt or in the range of or 0.9 % wt to 0.001 %wt or in the range of 0.9 % wt to 0.01 %wt or in the range of 0.8 % wt to 0.01 %wt or in the range of 0.8 % wt to 0.2 %wt in the range of 0.5 % wt to 0.001 %wt most preferably in the range of 0.1 % wt to 0.01 % wt (where the % wt is calculated with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined) nanoparticles as seeding material.

[0095] These embodiments all have the advantage that - as demonstrated in Fig. 1 - less nanoparticles are required than if the method for generating micro-particles as described in EP4154872 is employed.

[0096] In a preferred embodiment the method described herein in the first aspect is a method for generating micro-particles comprising the steps of c) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer wherein the concentration of the seeding material is in the range of 0.9 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of the active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined d) providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the same active ingredient in the range of 2 to 20, d) optionally isolating micro-particles.

[0097] Hence in another preferred embodiment the method described herein in the first aspect is a method for generating micro-particles comprising the steps of e) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), BHC243022-FC

[0098] -17- and at least one stabilizer wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of the active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined f) providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the same active ingredient in the range of 2 to 20, d) optionally isolating micro-particles.

[0099] Moreover, in another embodiment the method described herein in the first aspect is a method comprising the steps of a) Providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of active ingredientin the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of the same active ingredient in a crystallization medium wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the active ingredient in the range of 2 to 20, d) optionally isolating micro-particles.

[0100] Furthermore, in another embodiment the method described herein in the first aspect is a method comprising the steps of a) Providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, where the % wt is calculated with respect BHC243022-FC

[0101] -18- to the total concentration of the active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of the active ingredient in a crystallization medium wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 3 to 10, d) optionally isolating micro-particles

[0102] It is to be understood that the wording for step b) “b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a)” is meant to be identical to the wording “providing a homogenous saturated solution of the same active ingredient in a crystallization medium” in case the substance is an active ingredient.

[0103] In order to determine for a given substance e.g. an active ingredient which % wt concentration of nanoparticles e.g. active ingredient nanoparticles as seeding material can be used, simulations as described in Example 3a can be performed. Hence a person skilled in the art can model for a given compound which amount of seeding material is required for a desired particle size. In this respect it should be noted that also the values for nanoparticles e.g. active ingredient nanoparticles which are characterized by a nano-particle size in all of length, diameter or height of a d90 in the range of 10 nm to 999 nm can be used in the model as described in Example 3a instead of nanoparticles which are characterized by a size of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS) in at least one of length, diameter or height as well as a size of a d90 in the range of 1pm to 2pm as measured by Dynamic Light Scattering (DLS) in at least one of length, diameter or height as measured by Dynamic Light Scattering (DLS). In addition, the model as described in Example 3a can be used to determine the values for nanoparticles e.g. active ingredient nanoparticles which are of non-needle like appearance.

[0104] In an especially preferred embodiment the method for generating microparticles of the first aspect the substance is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide and the aqueous nanosuspension comprising nanoparticles as seeding material comprises a concentration in the range of 0.99 % wt to 0.001% wt (where the % wt is calculated with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous BHC243022-FC

[0105] -19- nanosuspension and the homogeneous saturated solution combined) nanoparticles as seeding material.

[0106] As shown in example 1 for (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide under the given conditions use of weight fractions of nanoparticles as seeding material in the range of 0.99% wt to 0.001% wt was advantageous since a concentration in the range of 0.99% wt to 0.001% wt less seeding material was required to generate microparticles with an average d90 value smaller than the microparticles obtained using the method of the art (cf. Table 1 and Table 2).

[0107] Thus, in a preferred embodiment of the method for generating micro-particles of the first aspect the method for generating micro-particles comprising the steps of a) Providing an aqueous nanosuspension comprising (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.99 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of (R)-2-(N-[4-amino-5- (4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in the aqueous nanosuspension and the homogeneous saturated solution combined, providing a homogenous saturated solution of (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in a crystallization medium wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising (R)- 2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 1 to 1000, d) optionally isolating micro-particles.

[0108] Additionally in another preferred embodiment of the method for generating micro-particles of the first aspect the method for generating micro-particles comprising the steps of a) Providing an aqueous nanosuspension comprising (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide nanoparticles as seeding material BHC243022-FC cocharacterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of (R)-2-(N-[4-amino-5- (4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in the aqueous nanosuspension and the homogeneous saturated solution combined, providing a homogenous saturated solution of (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in a crystallization medium wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising (R)- 2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 1 to 1000, d) optionally isolating micro-particles.

[0109] In a preferred embodiment of the method for generating microparticles described herein in the first aspect the supersaturation level is in the range of 2 to 20. In another preferred embodiment of the method for generating microparticles described herein in the first aspect the supersaturation level is in the range of 2 to 1000 or in the range of 2 to 100 or in the range of 2 to 50. In an even more preferred embodiment of the method for generating microparticles described herein in the first aspect the supersaturation level is in the range of 2 to 15. In an especially preferred embodiment of the method for generating microparticles described herein in the first aspect the supersaturation level is in the range of 3 to 10.

[0110] A person skilled in the art is aware of the fact that in a case where the mixing kinetics are faster than the dissolution no supersaturation has to occur during mixing of the homogenous saturated solution and the aqueous nanosuspension for controlled crystallisation to take place.

[0111] In a further preferred embodiment of the method for generating microparticles described herein in the first aspect the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles e.g. active ingredient nanoparticles as seeding material are provided at least one continuous fluid stream each. BHC243022-FC

[0112] -21-

[0113] Thus in this preferred embodiment the method for generating microparticles described herein in the first aspect comprises the steps of a) Providing at least one continuous fluid stream of an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano -particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing at least one continuous fluid stream of a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 1 to 1000 d) optionally isolating micro-particles.

[0114] In an especially preferred embodiment of the method for generating micro-particles of the first aspect the method for generating micro-particles comprising the steps of a) Providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.01 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 2 to 20. d) optionally isolating micro-particles. BHC243022-FC

[0115] -22-

[0116] In another especially preferred embodiment of the method for generating micro-particles of the first aspect the method for generating micro-particles comprising the steps of a) Providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 3 to 10, d) optionally isolating micro-particles.

[0117] In a highly preferred embodiment of the method for generating micro-particles of the first aspect the method for generating micro-particles comprising the steps of a) Providing an aqueous nanosuspension comprising (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in a crystallization medium BHC243022-FC

[0118] -23- wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 3 to 10, d) optionally isolating micro-particles.

[0119] In another highly preferred embodiment of the method for generating micro-particles of the first aspect the method for generating micro-particles comprising the steps of a) Providing an aqueous nanosuspension comprising (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide nanoparticles as seeding material characterized by a nano-particle size in all one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of a (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in a crystallization medium wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 3 to 10, d) optionally isolating micro-particles.

[0120] In a further especially preferred embodiment of the method for generating micro-particles of the first aspect the method for generating micro-particles comprising the steps of a) Providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d50 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, BHC243022-FC

[0121] -24- wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.01% wt, where the % wt is calculated with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 3 to 10, d) optionally isolating micro-particles.

[0122] In yet a further especially preferred embodiment of the method for generating micro-particles of the first aspect the method for generating micro-particles comprising the steps of a) Providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in all of length, diameter or height of a d50 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 3 to 10, d) optionally isolating micro-particles. BHC243022-FC

[0123] -25-

[0124] In still another further especially preferred embodiment of the method for generating microparticles of the first aspect the method for generating micro-particles comprising the steps of a) Providing at least one continuous fluid stream of an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in all of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing at least one continuous fluid stream of a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 3 to 10, d) optionally isolating micro-particles.

[0125] In an alternative especially preferred embodiment of the method for generating micro-particles of the first aspect the method for generating micro-particles comprising the steps of a) Providing at least one continuous fluid stream of an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano -particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing at least one continuous fluid stream of a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) BHC243022-FC

[0126] -26- wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 3 to 10, d) optionally isolating micro-particles.

[0127] In an alternative especially preferred embodiment of the method for generating micro-particles of the first aspect the method for generating micro-particles comprising the steps of a) Providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d50 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of the active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of the active ingredient in a crystallization medium wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 3 to 10, d) optionally isolating micro-particles.

[0128] In a most preferred embodiment of the method for generating micro-particles of the first aspect the method for generating micro-particles comprising the steps of a) Providing an aqueous nanosuspension comprising (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of (R)-2-(N-[4- amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in the aqueous nanosuspension and the homogeneous saturated solution combined BHC243022-FC

[0129] -27- b) providing a homogenous saturated solution of (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in a crystallization medium wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the (R)-2-(N-[4- amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in the range of 3 to 10, d) optionally isolating micro-particles.

[0130] In one embodiment of the method of the first aspect the provided aqueous nanosuspension comprises only nanoparticles e.g. active ingredient nanoparticles as seeding material characterized by a size of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS). In other words, the aqueous nanosuspension comprises only “symmetric” nanoparticles as seeding material which have a d90 in the range of 10 nm to 999 nm in every dimension of length, width and height.

[0131] In one embodiment the aqueous nanosuspension is supersaturated with respect to the substance e.g. the active ingredient constituting the nanoparticles as seeding material.

[0132] In another embodiment the homogenous saturated solution is supersaturated with respect to the substance .g. the active ingredient constituting the nanoparticles as seeding material in the aqueous nanosuspension, hence it is a homogenous supersaturated solution.

[0133] Thus in an embodiment the method for generating micro-particles of the first aspect comprises the steps of a) Providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 2 to 20, BHC243022-FC

[0134] -28- d) optionally isolating micro-particles wherein either the aqueous nanosuspension or the homogenous solution is supersaturated.

[0135] In one example of this embodiment the method for generating micro-particles of the first aspect comprises the steps of a) Providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS) and at least one stabilizer b) providing a homogenous solution of the same active ingredient in a crystallization medium c) mixing the homogenous solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 2 to 20, d) optionally isolating micro-particles wherein either the aqueous nanosuspension or the homogenous solution is supersaturated.

[0136] In another embodiment the homogenous saturated solution is undersaturated with respect to the substance constituting the nanoparticles as seeding material in the aqueous nanosuspension, hence it is a homogenous undersaturated solution.

[0137] Thus in another embodiment of the method for generating micro-particles of the first aspect the method for generating micro-particles comprises the steps of a) Providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous undersaturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane BHC243022-FC

[0138] -29- c) mixing the homogenous undersaturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 2 to 20, d) optionally isolating micro-particles.

[0139] Hence in one example of this embodiment of the method for generating micro-particles of the first aspect the method for generating micro-particles comprises the steps of

[0140] Providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of the active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined providing a homogenous undersaturated solution of the same active ingredient wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous undersaturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 2 to 20, d) optionally isolating micro-particles.

[0141] In an alternative embodiment of the method for generating micro-particles of the first aspect the provided aqueous nanosuspension comprises asymmetric seeding material characterized by i) a size of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS) in at least one of length, diameter or height as well as ii) size of a d90 in the range of 1pm to 2pm as measured by Dynamic Light Scattering (DLS) in at least one of length, diameter or height as measured by Dynamic Light Scattering (DLS).

[0142] Hence in one embodiment of the method for generating microparticles described herein in the first aspect the seeding material comprises asymmetric nano-particles e.g. active ingredient nanoparticles and the supersaturation level is in the range of 4 to 20, preferably in the range of 4 to 10.

[0143] Preferably the aqueous nanosuspension comprising nanoparticles as seeding material only comprises water as solvent and is termed aqueous nanosuspension. A person skilled in the art knows BHC243022-FC

[0144] -30- situations where it is appropriate to add components such as surfactants etc., however, an aqueous nanosuspension comprising nanoparticles as seeding material does not comprise organic solvents.

[0145] In a preferred embodiment of the method for generating microparticles described herein in the first aspect the aqueous nanosuspension comprising seeding material comprises at least one surfactant and at least one polymer i.e. a combination of stabilizers. For example the aqueous nanosuspension comprising seeding material comprises one surfactant and two polymers as stabilizers. In an alternative embodiment of the method described herein in the first aspect the aqueous nanosuspension comprising seeding material comprises only one polymer as stabilizer. In a further alternative embodiment of the method described herein in the first aspect the aqueous nanosuspension comprising seeding material comprises only one surfactant as stabilizer.

[0146] The at least one stabilizer may be selected from polymers that sterically and / or surfactants that electrostatically or sterically stabilizes the nano-particles. Polymers and surfactants known to a skilled person include:

[0147] • Alkyl celluloses, hydroxyalkyl celluloses, hydroxyalkylalkyl celluloses, carboxyalkyl celluloses, alkali metal salts of carboxyalkyl celluloses, carboxyalkylalkyl celluloses, carboxyalkyl cellulose esters, starches, pectins, chitin derivatives, polysaccharides, polyacrylic acid and its salts, poly methacrylic acid and its salts, polyvinyl alcohol, polyvinyl pyrrolidone, polyalkylene oxides, or a mixture of at least two of the above polymers. Preferably, the polymer is selected from methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose (HPC), hydroxybutylcellulose, hydroxyethylmethylcellulose, hydroxypropylmethylcellulose (HPMC), carboxymethylcellulose, sodiumcarboxymethylcellulose, carboxymethylethylcellulose, carboxyalkylcellulose ester, starches, sodium carboxymethylamylopectin, chitosan, dextran sulfate sodium salt, alginic acid, alkali metal and ammonium salts of alginic acid, carrageenans, galactomannans, tragacanth, agar-agar, gum arabic, guar gum, xanthan gum, polyacrylic acid and its salts, poly methacrylic acid and its salts, polyvinyl alcohol, polyethylene oxide, polypropylene oxide, copolymers of ethylene oxide and propylene oxide, N-vinylpyrrolidone-vinyl acetate copolymers or a mixture of at least two of the aforementioned polymers.

[0148] • Non-ionic surfactants in particular a poloxamer,

[0149] • a polyvinylpyrrolidone (PVP), in particular PVP K30, PVP K25, PVP K12 and polyvinylpyrrolidone-vinyl acetate copolymer (PVP VA), BHC243022-FC

[0150] -31- ionic surfactants such as anionic, cationic or zwitterionic (amphoteric) surfactants, wherein the ionic surfactant can be selected from acylamino acids (and salts thereof), such as: acylglutamates, for example sodium acylglutamate, di-TEA-palmitoylaspartate and sodium acaprylglutamate; acylpeptides, for example palmitoyl hydrolyzed milk protein, sodium cocoyl hydrolyzed soy protein and sodium / potassium cocoyl hydrolyzed collagen; Sarcosinates, for example, myristoyl sarcosine, TEA-lauroyl sarcosinate, sodium lauroyl sarcosinate and sodium cocoyl sarcosinate; Taurates, for example, sodium lauroyl taurate and sodium methyl cocoyl taurate; acyl lactylates, lauroyl lactylate, caproyl lactylate, alaninates; carboxylic acids and derivatives, such as: carboxylic acids, for example lauric acid, aluminum stearate, magnesium alkanolate and zinc undecylenate; ester carboxylic acids, for example calcium stearoyl lactylate and sodium PEG laurami de carboxylate; Ether carboxylic acids, for example sodium laureth carboxylate and sodium PEG cocamide carboxylate; phosphoric acid esters and salts, such as DEA-oleth-phosphate and dilaureth- phosphate; sulfonic acids and salts, such as acyl-isethionates, e.g. sodium / ammonium cocoyl isethionate, alkyl aryl sulfonates, alkyl sulfonates, for example sodium coco monoglyceride sulfate, sodium C-olefin sulfonate, sodium lauryl sulfoacetate and magnesium PEG cocamide sulfate, sulfosuccinates, for example dioctyl sodium sulfosuccinate, disodium laureth sulfosuccinate, disodium lauryl sulfosuccinate and disodium undecylenamido MEA sulfosuccinate; as well as sulfuric acid esters, such as alkyl ether sulfate, for example sodium, ammonium, magnesium, MIPA, TIPA laureth sulfate, sodium myreth sulfate and sodium C-pareth sulfate, alkyl sulfates, for example sodium, ammonium and TEA lauryl sulfate OR cationic surfactants, such as alkylamines, alkylimidazoles, ethoxylated amines, quaternary surfactant, quaternary ammonium compounds, and esterquats, wherein quaternary surfactants contain at least one N atom covalently bonded to 4 alkyl or aryl groups, which leads, independent of the pH value, to a positive charge. Advantageously alkyl betaine, alkyl amidopropyl betaine and alkyl amidopropyl hydroxysulfaine are used. Quaternary ammonium compounds are, in particular benzyltrialkylammonium chlorides or bromides, such as benzyldimethylstearylammonium chloride, further alkyltrialkylammonium salts, for example cetyltrimethylammonium chloride or bromide, alkyldimethylhydroxyethylammonium chlorides or bromides, dialkyldimethylammonium chlorides or bromides, alkylamidethyltrimethylammonium ether salts, alkylpyridinium salts, for example lauryl- or cetylpyrimidinium chloride, imidazoline derivatives and compounds of cationic character such as amine oxides, for example alkyldimethylamine oxides or alkylaminoethyldimethylamine oxides or cetyltrimethylammonium. BHC243022-FC

[0151] -32-

[0152] • Amphoteric surfactants can be Acyl / dialkyl ethylenediamines, for example sodium acylamphoacetate, disodium acylamphodipropionate, disodium alkylamphodiacetate, sodium acylamphohydroxypropyl sultonate, disodium acylamphodiacetate and sodium acylamphopropionate, and N-alkylamino acids, for example, aminopropylalkylglutamide, alkylaminopropionic acid, sodium alkylimidodipropionate and lauroamphocarboxy glycinate.

[0153] Preferred ionic surfactants include sodium dodecyl sulfate (SDS), sodium docusate (dioctyl sodium sulfosuccinate), sodium oleate and / or sodium deoxy cholate.

[0154] Employing an ionic surfactant has the advantage that it further stabilizes the nano -particles in the seeding material electrostatically,

[0155] Preferably the ionic surfactant is present in an amount in the range of 0.001 wt% to 10 wt% preferably 0.001 wt% and 0.4 wt% , most preferably in the range of 0.01 wt% to 0.2 wt% with respect to the components of the aqueous nanosuspension in suspension.

[0156] Preferably the at least one stabilizer is used in a ratio in the range of 1:1 to 10:1 (nanoparticle to stabilizer) preferably in a ratio of 10:3 . A skilled person knows whether to use only one or more than one stabilizer in a given situation.

[0157] In a preferred embodiment the combination of polymer and surfactant as stabilizer of the nanosuspension comprising seeding material consist of hydroxypropylmethylcellulose (HPMC) as polymer and sodium dodecyl sulfate (SDS) as surfactant.

[0158] In a preferred embodiment the combination of polymer and surfactants as stabilizer of the nanosuspension comprising seeding material consists of hydroxypropylcellulose (HPC) as polyvinylpyrrolidone (PVP) as non-ionic surfactant and sodium dodecyl sulfate (SDS) as ionic surfactant.

[0159] In the most preferred embodiment the method described herein in the first aspect the aqueous nanosuspension comprising seeding material comprises a combination of polyvinylpyrrolidone (PVP) KI 2, hydroxypropyl cellulose (HPC) und dodecyl sulfate (SDS).

[0160] In one example of this embodiment the aqueous nanosuspension comprising nanoparticles as seeding material e.g. active ingredient nanoparticles comprises HPC in the range of 2wt% to 3wt%, BHC243022-FC

[0161] -33- poly vinylpyrrolidone (PVP) KI 2 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0.1wt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10:1 to 1: 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2: 1 to 1:2. In a further example of this embodiment the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) K12 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0.1 wt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1: 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2: 1 to 1:2., wherein the substance is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide, wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane and wherein the supersaturation level with respect to the substance in the range of 3 to 10.

[0162] In a preferred embodiment of the method for generating microparticles described herein in the first aspect the mixing time < growth time.

[0163] In a preferred embodiment of the method for generating microparticles described herein in the first aspect mixing is continued until a d90 of > 1pm is reached.

[0164] The supersaturation level can be adjusted by the amount of the homogenous solution which can be undersaturated, saturated or supersaturated, and the amount of aqueous nanosuspension comprising nanoparticles as seeding material.

[0165] In one embodiment of the method for generating microparticles described herein in the first aspect step c) of mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material starts in at least one mixing element (3a, 3b).

[0166] The at least one mixing element (3a, 3b) can be a T- or Y-shaped solid or flexible tubing from different materials (e.g. metal, glass, ceramics, polymers) or can be a more elaborated component that contains active elements like e.g. rotating components or static elements e.g. like lamella or baffles with the effect of increasing the mixing efficiency. Acoustic mixing technology can also be applied for efficient mixing and mixing could also be accomplished by impinging jet technology or even with appropriate elements of micro reaction technology.

[0167] In one embodiment of the method for generating microparticles described herein in the first aspect the homogenous solution which can be undersaturated, saturated or supersaturated and the aqueous BHC243022-FC

[0168] -34- nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element (3a, 3b) at the same time.

[0169] In a preferred embodiment of the method for generating microparticles described herein in the first aspect the homogenous solution which can be undersaturated, saturated or supersaturated and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element (3a, 3b) as at least one continuous fluid stream each at the same time.

[0170] In a further preferred embodiment of the method for generating microparticles described herein in the first aspect the homogenous solution is a homogenous saturated solution and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element (3a, 3b) at the same time.

[0171] Moreover, in a preferred embodiment of the method for generating microparticles described herein in the first aspect the homogenous solution is a homogenous saturated solution and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element (3a, 3b) as at least one continuous fluid stream each at the same time.

[0172] In detail the level of supersaturation in this embodiment can be adjusted by

[0173] 1) the concentration of the active ingredient in the homogenous solution which can be undersaturated, saturated or supersaturated. The higher the concentration is the higher is the supersaturation.

[0174] 2) the ratio of the homogenous solution, which can be undersaturated, saturated or supersaturated, relative to the aqueous nanosuspension comprising seeding material. The ratio of both can also be expressed as the ratio of both flow rates. The higher the flow rate of the aqueous nanosuspension comprising seeding material relative to the homogeneous solution, the higher is the supersaturation.

[0175] 3) the temperature at which the homogenous supersaturated solution and aqueous nanosuspension comprising seeding material enter a mixing element. The lower the temperature in the mixing element the higher is the supersaturation therein.

[0176] All here described options for adjusting the supersaturation in the mixing element can be combined with each other since they can be individually adjusted.

[0177] In a preferred embodiment the at least one mixing element is a T- or Y-shaped tubing. BHC243022-FC

[0178] -35-

[0179] Hence in one embodiment of the method described herein in the first aspect said method is a method is comprising the steps of a) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined and at least one stabilizer b) providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material wherein the mixing starts in mixing element and said mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time, d) optionally isolating micro-particles wherein in step c) the supersaturation level with respect to the active ingredient is in the range of 2 to 20

[0180] Preferably the ratio of the flow rates at which the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles e.g. active ingredient nanoparticles as seeding material are provided to the at least one mixing element is in the range of 2:1 to 1:2.

[0181] Moreover, in a preferred embodiment of the of the method for generating microparticles described herein in the first aspect the ratio of the flow rates at which the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles e.g. active ingredient nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time is in the range of 2: 1 to 1 :2.

[0182] Even more preferably the ratio of the flow rates at which the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles e.g. active ingredient nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time is 1: 1. BHC243022-FC

[0183] -36-

[0184] Hence, in another preferred embodiment of the of the method for generating microparticles described herein in the first aspect the ratio of the flow rates at which the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles e.g. active ingredient nanoparticles as seeding material are provided to the at least one mixing element (3 a, 3b) as at least one continuous fluid stream each at the same time is 1: 1.

[0185] Thus in an especially preferred embodiment of the method for generating microparticles described herein in the first aspect the method comprises the steps of a) Providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time. d) optionally isolating micro-particles.

[0186] In a further embodiment of the method described herein in the first aspect said method is a method is comprising the steps of a) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer b) providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the BHC243022-FC

[0187] -37- homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time, d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the active ingredient is in the range of 3 to 5 wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) KI 2 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0.1 wt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1: 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2:1 to 1:2. wherein the active ingredient is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4- fluoro-anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane.

[0188] In a further embodiment of the method described herein in the first aspect said method is a method is comprising the steps of a) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer b) providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time in a ratio of the flow rates of homogenous saturated solution to aqueous nanosuspension comprising nanoparticles as seeding material in the range of 2: 1 to 1:2 d) optionally isolating micro-particles. BHC243022-FC

[0189] -38- wherein in step c) the supersaturation level with respect to the active ingredient is in the range of 3 to 5 wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) KI 2 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0.1 wt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1 : 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2:1 to 1:2. wherein the active ingredient is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4- fluoro-anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane.

[0190] In a highly preferred embodiment of the method described herein in the first aspect said method is a method is comprising the steps of a) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer b) providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time in a ratio of the flow rates of the homogenous saturated solution to aqueous nanosuspension comprising active ingredient nanoparticles as seeding material of 1 : 1 , d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the active ingredient is in the range of 3 to 5, wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, BHC243022-FC

[0191] -39- wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) KI 2 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0.1 wt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1: 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2:1 to 1:2. wherein the active ingredient is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4- fluoro-anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane.

[0192] In another highly preferred embodiment of the method described herein in the first aspect said method is a method is comprising the steps of a) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer b) providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time in a ratio of the flow rates of the homogenous saturated solution to aqueous nanosuspension comprising active ingredient nanoparticles as seeding material of 1 : 1 , d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the active ingredient is in the range of 3 to 5, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) KI 2 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0.1 wt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range BHC243022-FC

[0193] -40- of 10: 1 to 1: 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2:1 to 1:2. wherein the active ingredient is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4- fluoro-anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane.

[0194] In a further preferred embodiment of the method for generating microparticles described herein in the first aspect the method comprises the steps of a) Providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time. d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the substance is in the range of 3 to 10.

[0195] Preferably step c) hence starts in the at least one mixing element (3a, 3b) when the aqueous nanosuspension and the homogeneous saturation solution come into contact with each other and the growth of the particles starts. If the at least one mixing element (3 a, 3b) comprises a discharge channel crystallisation may continue in said discharge channel and ends when the supersaturation BHC243022-FC

[0196] -41- in the mixture is reduced due to the particle growth or the generated microsuspension leaving the at least one mixing element (3a, 3b). The microsuspension can be filtered directly when exiting the at least one mixing element or can be collected in a collecting vessel (4), which can optionally also contain pure antisolvent as a "quencher", for further processing.

[0197] Alternatively the step c) starts and takes place in the at least one mixing element.

[0198] Therefore what is also disclosed herein relates to a method for generating microparticles comprising the steps of a) Providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined in at least one first storage vessel (1) b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane in at least one second storage vessel (2) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 3 to 10 via providing the homogenous saturated solution and the aqueous nanosuspension to at least one mixing element (3a, 3b) and said at least one mixing element (3a, 3b) is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time., d) collecting the suspension generated in step c) in at least one collection vessel (4) e) optionally isolating micro-particles.

[0199] In a preferred embodiment of this method, the method for generating microparticles comprises the steps of BHC243022-FC

[0200] -42- a) providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time, d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the substance is in the range of 3 to 5 wherein the concentration of the seeding material is in the range of 0.1 % wt and a concentration of 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) K12 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0. lwt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1 : 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2: 1 to 1:2. wherein the substance is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane.

[0201] In a further preferred embodiment of this method, the method for generating microparticles comprises the steps of a) providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer BHC243022-FC

[0202] -43- b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element at the same time in a ratio of the flow rates of homogenous saturated solution to aqueous nanosuspension comprising nanoparticles as seeding material in the range of 2: 1 to 1 :2 d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the substance is in the range of 3 to 5 wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) K12 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0. lwt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1 : 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2: 1 to 1:2. wherein the substance is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane.

[0203] In an even more preferred embodiment of this method the method for generating microparticles comprises the steps of a) providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material wherein the mixing starts in at least one mixing element BHC243022-FC

[0204] -44- and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time in a ratio of the flow rates of homogenous saturated solution to aqueous nanosuspension comprising nanoparticles as seeding material of 1: 1 d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the substance is in the range of 3 to 5 wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) K12 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0. lwt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1 : 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2: 1 to 1:2. wherein the substance is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane.

[0205] In another preferred embodiment of this method the method for generating microparticles comprises the steps of a) providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time in a ratio of the flow rates of homogenous saturated solution to aqueous nanosuspension comprising nanoparticles as seeding material of 1: 1 BHC243022-FC

[0206] -45- d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the substance is in the range of 3 to 5 wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) K12 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0. lwt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1 : 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2: 1 to 1:2. wherein the substance is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane.

[0207] In an alternative embodiment of this method the method for generating microparticles comprises the steps of a) providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time, in a ratio of the flow rates of homogenous saturated solution to aqueous nanosuspension comprising nanoparticles as seeding material of 1: 1 d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the substance is in the range of 1 to 10 wherein the concentration of the seeding material is in the range of 1,9 % wt to 2,1% wt, wherein the substance is indomethacin. BHC243022-FC

[0208] -46-

[0209] Typical mixing times t mix are in the range of 10 ms < t mix < 10s with mixing time defined as the residence time in the at least one mixing element.

[0210] As mentioned above in a case where the mixing kinetics are faster than the dissolution no supersaturation has to occur during mixing of the homogenous saturated solution and the aqueous nanosuspension for controlled crystallisation to take place.

[0211] The growth of the nanoparticles into microparticles can also be controlled via the length of the discharge channel and the flow rate in it.

[0212] In a further embodiment of the method described herein in the first aspect step c) takes place in more than one mixing element (3a,b) and the at least one second mixing element (3b) is located in series to the at least one first mixing element (3a). Preferably the further homogenous saturated solution is added to the second mixing element (3b). This embodiment, which is depicted in Fig. 3 has the advantage that the crystals of the substance can further grow if a larger size is desired. A person skilled in the art understands that the set-up of Fig. 1-3 can also be different e.g. via having several vessels with the same content while still achieving the same overall process.

[0213] In a further embodiment of the method described herein in the first aspect the at least one mixing element (3a, 3b) comprises one or more components selected from the group consisting of baffles, single or multiple nozzle elements, lamellas, jet disperser elements ("impinging jet") or active elements such as stirrers or similar to facilitate efficient mixing of the homogenous saturated solution and the aqueous nanosuspension. The aim of the components is always to achieve efficient (rapid) mixing so that the nanoparticles are distributed very quickly and homogeneously in order to keep the distance the active ingredient molecules of the homogenous supersaturated solution have to diffuse to the nearest nanoparticle as short as possible and thus minimize the probability of spontaneous nucleation.

[0214] In a further embodiment of the method described herein in the first aspect the method relates to a process for generating micro-particles comprising the further step prior to step a) of generating the aqueous nanosuspension comprising nanoparticles.

[0215] For example the nanoparticles of the nanosuspension are generated in situ directly before mixing with the homogenous supersaturated solution (as exemplary depicted in Fig, 4) . Thus, the use of a separately prepared nanosuspension is not necessary which saves a whole process step. This process BHC243022-FC

[0216] -47- is in a certain sense similar to a classical crystallization, in which the microparticles are generated by spontaneous nucleation and growth in a supersaturated drug solution by variation of concentration and temperature. However, according to the method of the first aspect described herein, nucleation and growth are spatially separated from each other and can be controlled separately via flow rate, temperature and / or concentration. As a result, a narrower size distribution can be expected compared to classical crystallization. A further advantage of this embodiment is that those in situ generated nanoparticles need less stabilizers since they are directly used as seeds within a few seconds after generation. No long-time stabilization of the nanosuspension is needed in this case.

[0217] In a further embodiment the nanoparticles are not produced by wet -milling in an aqueous medium in order to obtain an aqueous nanosuspension, but wet-milling is carried out in a non-aqueous (organic) liquid in which the active ingredient is not or only slightly soluble in order to obtain an organic nanosuspension. This milling process is preferably carried out if the active ingredient has a sufficient solubility in water so that aqueous milling is not possible since the material would mainly dissolve. Another case where this method may be preferred is when wet milling in aqueous media leads, for example, to hydrates which are not wanted as nanoseeds for later crystal growth. In this case, suitable stabilizers are dissolved in the organic medium before milling which stabilize the (organic) nanosuspension. The microcrystals are then produced in an analogous way as in the previously mentioned embodiments. It must always be considered that the term solvent refers to the liquid in which the active ingredient is dissolved and the term antisolvent refers to the liquid in which the nanoparticles are dispersed.

[0218] What is described herein also relates to a method for generating micro-particles comprising the steps of a) providing a homogenous saturated solution of a substance in a crystallization medium b) providing a seeding material characterized by i) a size of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS) in at least one of length, diameter or height as well as ii) size of a d90 in the range of 1pm to 2pm as measured by Dynamic Light Scattering (DLS) in at least one of length, diameter or height as measured by Dynamic Light Scattering (DLS), comprising at least one stabilizer, wherein said seeding material is of the same substance as the substance of the homogenous saturated solution of step a) c) bringing the homogenous saturated solution in contact with the seeding material, d) optionally isolating micro-particles. BHC243022-FC

[0219] -48-

[0220] Hence in this embodiment of the method the seeding material does not have to be provided as nanosuspension but could also be provided as tablet or powder.

[0221] The seeding material can be obtained by the method as described in WO 2021 / 069350 i.e. via a method comprising the steps of:

[0222] 1) suspending a pharmaceutical active substance in an aqueous solution of a polymer,

[0223] 2) milling the suspension comprising the pharmaceutical active substance until the pharmaceutical active substance particles have a d90 value in the particle size distribution of < 1 pm and

[0224] 3) drying the mixture obtained in step 1); characterized in that before step 2) the pharmaceutical active substance is further contacted with an ionic surfactant and wherein the polymer and the surfactant are present in a relative weight ratio of > 40: 1 to < 100: 1.

[0225] The seeding material obtained by the method of WO 2021 / 069350 as described above provided as powder or as tablet has the advantage that it can be stored more convenient than seeding material suspended in crystallization medium. Preferably, the nano-particles are obtained via nano-grinding thereby providing a seeding material characterized by a nano-particle size of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy comprising at least one stabilizer.

[0226] In a second aspect what is described herein relates to a) Providing a saturated aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is not the same substance as the substance in the seeding material of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 1 to 1000 d) optionally isolating micro-particles

[0227] Hence in aspect b) crystal growth is initiated not by homogeneous nucleation as described above, but by heterogeneous nucleation. In this case, crystal growth of the substance (e.g. the active ingredient) is achieved by adding a suspension containing nanoparticles or nanoseeds, respectively, BHC243022-FC

[0228] -49- of a different material. In this case, the nanomaterial would preferably but not necessarily consist of an inert and nontoxic inorganic material, such as oxides like silica or carbonates.

[0229] What is described herein relates to a method for generating micro-particles comprising the steps of a) providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 1 to 1000, d) optionally isolating micro-particles.

[0230] The above method, wherein the substance is an active ingredient.

[0231] The above method wherein the aqueous nanosuspension comprising nanoparticles as seeding material comprises a concentration in the range of 0.99 % wt to 0.001% wt nanoparticles as seeding material.

[0232] The above method wherein the substance is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2- yl]-4-fluoro-anilino)propanamide and the aqueous nanosuspension comprising nanoparticles as seeding material comprises a concentration in the range of 0.99 % wt to 0.001% wt nanoparticles as seeding material.

[0233] The above method comprising the steps of a) Providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) BHC243022-FC

[0234] -50- wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 2 to 20, d) optionally isolating micro-particles.

[0235] The above method comprising the steps of a) Providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d50 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 3 to 10, d) optionally isolating micro-particles.

[0236] The above method comprising the steps of a) Providing an aqueous nanosuspension comprising (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of i.e. substance e.g. active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined BHC243022-FC

[0237] -51- b) providing a homogenous saturated solution of (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol- 2-yl]-4-fluoro-anilino)propanamide in a crystallization medium wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 3 to 10, d) optionally isolating micro-particles.

[0238] The above method, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) KI 2 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0. lwt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1: 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2: 1 to 1:2. wherein the substance is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane and wherein the supersaturation level with respect to the substance in the range of 3 to 10.

[0239] What is described herein also relates to a method comprising the steps of a) providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material wherein the mixing starts in mixing element and said mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time, BHC243022-FC

[0240] -52- d) optionally isolating micro-particles.

[0241] The immediately above method comprising the steps of a) providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material wherein the mixing starts in mixing element and said mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time, d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the substance is in the range of 1 to 1000.

[0242] The last above method comprising the steps of a) providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time, d) optionally isolating micro-particles. BHC243022-FC

[0243] -53- wherein in step c) the supersaturation level with respect to the substance is in the range of 3 to 5 wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) KI 2 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0. lwt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1:10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2:1 to 1:2. wherein the substance is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane.

[0244] The last above method comprising the steps of a) providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time in a ratio of the flow rates of homogenous saturated solution to aqueous nanosuspension comprising nanoparticles as seeding material in the range of 2:1 to 1:2 d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the substance is in the range of 3 to 5 wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) KI 2 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0. lwt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1:10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2:1 to 1:2. BHC243022-FC

[0245] -54- wherein the substance is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane.

[0246] The last above method comprising the steps of a) providing an aqueous nanosuspension comprising nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer b) providing a homogenous saturated solution of a substance in a crystallization medium where said substance is the same substance as the substance constituting the nanoparticles in the nanosuspension of a) c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time in a ratio of the flow rates of the homogenous saturated solution to aqueous nanosuspension comprising nanoparticles as seeding material of 1: 1, d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the substance is in the range of 3 to 5, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) KI 2 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0. lwt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1:10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2:1 to 1:2. wherein the substance is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane. BHC243022-FC

[0247] -55-

[0248] EXAMPLES

[0249] The various aspects of the subject-matter described in this application are illustrated by the following examples which are not meant to be limiting in any way. Instead the examples merely serve to illustrate the subject-matter described herein.

[0250] Example 1: Comparison of nano seeding via cooling crystallisation with the method described herein

[0251] Preparation of starting materials:

[0252] Preparation of [4-amino-2-(4-fluoroanilino)thiazol-5-yl]-(4-methoxyphenyl)methanone:

[0253] A mixture of l-fluoro-4-isothiocyanato-benzene (300 g, 1.96 mol) and tri ethylamine (297 g, 2.94 mol, 1.5 equiv) in acetonitrile (0.90 L) was warmed to 60 °C. To this mixture was added a solution of cyanamide (90.6 g, 2.15 mol, 1.1 equiv) in acetonitrile (1.35 L) within 1 h. After an additional 1 h, a solution of 2-bromo-l-(4-methoxyphenyl)ethanone (449 g, 1.96 mol, 1.0 equiv) in acetonitrile (2.25 L) was added at 60 °C within 75 min. After an additional 15 min, the suspension was cooled to 20 °C within 0.5 h and stirred at that temperature for an additional 0.5 h. The mixture was filtered, the precipitate was washed with H2O and dried at 50 °C in vacuum to yield [4-amino-2-(4- fluoroanilino)thiazol-5-yl]-(4-methoxyphenyl)methanone (606 g, 1.75 mol, 89% yield, 99.0% purity) as a yellow to orange solid.

[0254] Preparation of (2S)-2-bromopropanamide:

[0255] A mixture of (2S)-2-bromopropanoic acid (200 g, 1.31 mol, 88% ee, purchased from ABCR) and thionyl chloride (218 g, 1.83 mol, 1.4 equiv) were heated to 50 °C. After 18 h, the resulting crude acyl chloride was diluted with 2-methyltetrahydrofuran (1.85 L) and added to 30% aqueous ammonia solution (381 g, 3.27 mol, 2.5 equiv) at -15 to 7 °C within 75 min. After completion of the addition, the temperature was raised to 20 °C and water (200 mL) was added. The layers were separated and the aqueous layer was extracted with 2-methyltetrahydrofuran. The combined organic layers were washed with saturated aqueous sodium chloride solution and part of the solvent was removed in vacuo at 40 °C. n-Heptane was added, the resulting suspension was cooled to 0-5 °C, kept at that temperature for 1 h, filtered, and the precipitate was washed with n-heptane. Drying of the filter cake in vacuum at 35 °C afforded (2S)-2-bromopropanamide (147 g, 967 mmol, 74% yield, 99.65% purity, 95% enantiomeric excess) as a colourless solid.

[0256] Preparation of (2R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino) propanamide BHC243022-FC

[0257] -56-

[0258] A suspension of [4-amino-2-(4-fluoroanilino)thiazol-5-yl]-(4-methoxyphenyl)methanone (120 g, 349 mmol), (2S)-2-bromopropanamide (63.7 g, 419 mmol, 1.2 equiv, 95% enantiomeric excess) and potassium phosphate (K3PO4, 148 g, 699 mmol, 2.0 equiv) in acetonitrile (1200 mL) was heated to 60 °C and stirred at that temperature for 1 h. The temperature was subsequently lowered to 20 °C within 40 min, and water (1200 mL) was simultaneously added within 20 min. The resulting triphasic mixture was stirred at 20 °C for 3 h, filtered, and the filter cake was washed with water (3 x 240 mL), affording (2R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide as a light yellow solid (124 g wet, 99.14 % purity, 99.2% enantiomeric excess). The water content of the wet filter cake was determined and found to be 16%, corresponding to 20 mL of residual water in the wet filter cake. The wet filter cake was dissolved in a mixture of acetone (1350 mL) and water (130 mL) at ambient temperature, resulting in a solution with a small amount of residual solid. The mixture was filtered, and the filtrate was added to water (2000 mL) at 4-10 °C within 38 min. The resulting disperse, off-white suspension was filtered and the filter cake was washed with water (2 x 150 mL) and dried in vacuo at 50 °C, furnishing (2R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide (90.3 g, 216 mmol, 62% yield, 99.12% purity, 99.3% enantiomeric excess) as an off-white, lumped powder. The chemical purity and enantiomeric excess were determined as specified above according to HPLC methods 4 and 5.

[0259] Method HPLC4: Chemical purity.

[0260] Equipment: High-performance liquid chromatograph (with a pressure range of up to 600 bar and a dwell volume of approx. 850 pL) with a thermostatically controlled column oven, UV detector and data evaluation system (e.g. Agilent 1260)

[0261] C olumn : Waters A equity BEH Shield

[0262] C18 Length: 50 mm, inner diameter: 2.1 mm, particle size: 1. 7 pm Maximum pressure: 600 bar Typical starting pressure: 500 bar

[0263] Eluents: A: 1580mg Ammonium bicarbonate + 80pL formic acid (98+%) / IL Milli-Q water (compressibility: Use Solvent Types) BHC243022-FC

[0264] -57-

[0265] B: Acetonitrile (compressibility: Use Solvent Types)

[0266] Gradient: Time (min) B (%)

[0267] 0.00 5.0

[0268] I.00 5.0

[0269] 4.50 45.0

[0270] 7.50 60.0

[0271] 8.50 80.0

[0272] 9.50 80.0

[0273] 10.00 5.0

[0274] I I.00 5.0

[0275] The percentage of mobile phase A is the difference between B (%) and 100%.

[0276] Equilibration time: 1 min (at starting condition)

[0277] Flow rate: 0.7 mL / min

[0278] Draw speed: 200 pL / min

[0279] Injection volume: 2.5 pL (RT)

[0280] Needle wash: Flush port period (7 sec)

[0281] Solvent: Methanol

[0282] Column temperature: 40 °C

[0283] Data sampling rate: 20 Hz (> 0.013 min (0.25 s response time)

[0284] Detection wavelength: 270 nm, band width: 4 nm

[0285] Margin for negative 100 mAU absorbance:

[0286] Detector cell path: 10 mm

[0287] Slit: 4 nm

[0288] Sample solvent: Acetonitrile

[0289] Sample / Calibration Dissolve sample at a concentration of approx. 0.25 mg / mL (e.g. weigh exactly solution: 25 mg and dissolve in 100 mL) of the substance with sample solvent and fill up to the calibration mark.

[0290] Method HPLC5:

[0291] Equipment: Ultrahigh-performance liquid chromatograph with a thermostatically controlled column oven, UV detector and data evaluation system, dwell volume approx. 200 pL (e.g. Agilent 1290)

[0292] Column : Daicel Chiralpak IB N-3

[0293] Length: 150 mm, inner BHC243022-FC

[0294] -58- diameter: 4.6 mm, particle size: 3.0 pm

[0295] Maximum pressure: 300 bar

[0296] Typical starting pressure: 210 bar

[0297] Column temperature: 25 °C

[0298] Mobile phase: A: n-Heptane / Ethanol+ 0.1 % TFA (50 % + 50 %; V:V) (compressibility: use Solvent-Type)

[0299] Flow rate 1.50 mL / min

[0300] Isocratic: A (%): 100

[0301] Runtime: 8.0 min

[0302] Detector cell path: 10 mm

[0303] Detection wavelength: 265 nm, band width: 4 nm

[0304] Data sampling rate: 2.5 Hz (> 0.1 min (2 s response time))

[0305] Margin for negative 100 mAU absorbance:

[0306] Slit: 4 nm

[0307] Injection volume: 3.0 pL (RT)

[0308] Draw speed: 200 pL / min

[0309] Needle wash: Flush port period (7 sec)

[0310] Solvent: Ethanol

[0311] Sample solvent: Ethanol

[0312] Sample solution: Dissolve sample at a concentration of approx. 1.0 mg / mL (e.g. weigh exactly 25 mg and dissolve in 25 mL) of the substance with sample solvent and fill up to the calibration mark. a) Nano seeding via cooling crystallisation (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide powder (essentially anhydrous, prepared according to the protocol described in above (180 g) was added to a mixture of acetone (877 g) and water (175.5 g) and heated up to 55 °C, resulting in a solution with a small amount of residual solid. The mixture was filtered with a K300 20 pm filter cloth, and the filtrate was cooled down to 35 °C in 1 hour. A nanosuspension consisting of 79.8 wt% water, 10 wt% (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide, 8 wt% PVP K12 (Acros Organics), 2 wt% HPC (Klucel ELF / Ashland and 0.2 wt% SDS (abcr) had been previously prepared via milling in a Buhler PML2 (small milling chamber) with a bead size of 0.1 mm (Yttrium stabilized Zirconium Oxide) for 40 min at 3000 rpm and a maximum temperature of 35°C thereby obtaining suspended nano-particles as nanosuspension. 35 g of said nanosuspension BHC243022-FC

[0313] -59- were added as seeding material. Subsequently the suspension was cooled down to 20 °C in 1 hour. To increase the yield, 1165.5 g of water was continuously added over 1 hour with a subsequent stirring time of 1 hour. The resulting disperse, white suspension was filtered in a pressure filter and a PP 2703 filter cloth. The filter cake was washed with water (2 x 800 mL) and dried in vacuum at 50 °C, furnishing (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide (164.55 g, 91.4% yield, 99.59% purity, 99.74% enantiomeric excess) as an white, lumped powder. The result is shown in Fig. 1. b) Novel Method

[0314] At first 100 ml of the homogenous supersaturated solution and 100 ml of the nanosuspension comprising nanoparticles were prepared using (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2- yl]-4-fluoro-anilino)propanamide as active ingredient. To prepare the homogenous supersaturated solution the solvent for (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide was prepared first via mixing 80 g acetone and 20 g water. Then 8,8 g (R)-2- (N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide powder synthesized as described above was dispersed (magnetic stirring) inside the prepared water-acetone solvent and left equilibrating for 2 hours, after which particles could still be detected. This suspension was then filtered with a syringe in a millipore filter (0.22 pm, hydrpholic, MCE), to separate the residual particles from homogenous saturated solution. It had been determined previously that the (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide content of a saturated solution was 4.4 wt% using the gravimetric method as detailed and explained in Beckmann 2013 pages 64 and 65.)

[0315] For preparation of the aqueous nanosuspension comprising nanoparticles as seeding material 10 wt% (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide microcrystals were dispersed in demineralized water in the presence of 8 wt% PVPK12, 2 wt% HPC and 0.2 wt% SDS. resulting in 60 g suspension with 10 wt% (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide microparticles. The suspension was milled in stirred media mill (Picoline by Hosoqawa-alpine) with 90 ml milling chamber. 80% of milling chamber volume was filled with 0.4-0.6 mm grinding beads made of yttrium stabilized zircon oxide. The milling chamber was filled completely with suspension (air free). Milling time was 60 min below 30°C and 2500 rpm. The resulting aqueous nanosuspension comprising nanoparticles as seeding material (10wt% API, 8 wt% PVPK12, 2 wt% HPC and 0.2 wt% SDS) was diluted to 0.198 wt% (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide nanoparticles in water i.e. 0.198 wt% in relation to the total amount of (R)-2- (N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in the homogenous supersaturated solution with an (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- BHC243022-FC

[0316] -60- anilino)propanamide content of 4.4 wt% prepared as described above and the aqueous nanosuspension combined,

[0317] In the next step a continuous fluid stream of the prepared homogenous saturated solution with respect to (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide was mixed with a continuous fluid stream of the prepared, diluted aqueous nanosuspension comprising (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide. In order to do so 45g of (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide homogenous supersaturated solution and 45g of aqueous nanosuspension were provided, which had been prepared as described above in 100 ml Schott-bottles, which were stirred on a magnetic stirrer (IKA RH basic 2). Both bottles were connected to pumps (SyrDos by HiTEC ZANG) and a mixing element here a T-shaped mixer with PTFE tubes with inner diameter of 1 mm. The T-shaped mixer was 3D printed from Teflon and was characterized by 1mm internal diameter, 1.5 x 1.5 cm horizontal and vertical T-sides. The flow rates were set to 20 ml / min i.e. both the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material entered the T-mixer at the same time in a 1 : 1 ratio. Below the T-mixer a stirred (magnetic stirrer IKA RH basic 2) Schott bottle was placed to collect and homogenize the crystallized suspension. The suspension was flowing directly from T-mixer and via a silicone tube with inner diameter of 1.6 mm into the Schott bottle. The experiment was carried out at room temperature. Samples were taken directly after mixing and after 60 min stirring in the Schott bottle. The final crystallized suspension was filtered with a standard Buchner filtration.

[0318] A supersaturation level in the range of around 4,4 was reached in T-mixer (this was the maximum attainable supersaturated level). The maximum attainable supersaturation was calculated as the ratio between the concentration of (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide in the joint streams and the solubility of the (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in the suspension comprising microparticles resulting from the mixing (with a measured value of 0.01 g / gsoiution for a solvent composition of 50 / 50 wt % acetone / water). The composition cout in the joint streams was calculated based on the mass balance on the T-mixer as follows: cOut=(Qm,i*Cm,i + Qm,2*Cin,2) / (Qout). Hence for this example as the concentration of (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4- fluoro-anilino)propanamide in the joint streams was 0.044 g / gsoiution and the solubility of the (R)-2- (N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in the solvent mixtures resulting from the mixing was 0.01 g / gsoiution the maximum attainable supersaturation was calculated to be 4.4 (0.044 / 0.01). BHC243022-FC

[0319] -61-

[0320] The resulting particles and corresponding particle size distributions were measured via laser diffraction using a Mastersizer 3000 with the settings: water dispersion unit, 2500 rpm stirrer speed, Mie-theory, refractive index 1.63). The results given in Fig. 1 demonstrated that less nanoseeds - i.e. a concentration of the seeding material below 1% wt - could be used to reach smaller particle sizes via the method described herein in comparison to methods described in the art . lc) Comparison Experiment

[0321] For comparison also an experiment with the settings of lb) but with a nanoparticle concentration close to zero (10‘8%wt) was carried out. As seen in Fig. 6 the resulting particles were larger if almost no seeding material was present. ld) Experiment using different active ingredient

[0322] The experiments la and lb were repeated with the substance indomethacin but with a different concentration of nanoseeds in the aqueous nanosuspension. The results are given in Fig. 7. Moreover, it was found for indomethacin that via using the method described herein it was possible to generate the gamma polymorph (data not shown). This was surprising since the unstable alpha polymorph is generated if antisolvent crystallisation as described in the art (da Silva et al. Crystal Growth & Design Vol 22 / Issue 10, 2022) is used for indomethacin. Hence, the method described herein is in addition to the advantages described above favourable for a substance generating at least two polymorphs where the undesirable polymorph exhibits the faster nucleation and thus grows faster than the desired polymorph.

[0323] Example 2 Modelling

[0324] In order to facilitate the determination of the amount of seeding material needed to generate the same small particle size distribution when using the method described herein a model was set up. This model of Example 2 was designed to model example la, however, a person skilled in the art can readily use it for other substances. In this example the assumed nanoparticles of the aqueous nanosuspension comprising nanoparticles as seeding material used in a standard population balance equation (see for instance D. Ramkrishna, Population Balances, Theory and Applications to Particulate Systems in Engineering, 2000) were characterized by a size of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS) in all of length, diameter or height. One characteristic length was used. The particles were described in the model by a simplified geometrical shape, namely as a square-based prism, whose height was bigger than its width as this corresponds to the shape of (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide nanoparticles. Using the aforementioned geometry, the crystals volume and surface can be computed as function of the characteristic length times a volume shape factor (kv = BHC243022-FC

[0325] -62-

[0326] 0.112) and a surface shape factor (ks = 1.556), respectively. The values for the shape factors have been derived assuming for all particles a constant Aspect Ratio (AR, defined as the ratio between the needle length and the needle width) value of 3. In this exemplary simulation the length of the needles or the height of the prisms was used and had a d90 value in range of 10-999 nm. The other dimensions had an d90 value in range of 10-999 nm. The population balance equation was coupled with the standard mass balance on the active ingredient, relating the crystal mass via the second moment to the free solute to account for the dynamic variation of supersaturation. The high- resolution finite volume method (with Koren flux limiter) was implemented in Python, according to the method reported by Zhang et al. (Comp & Chem Eng., 108612-183, 2024), and a nonlinear grid (log-scaled) was chosen to simulate the system, providing a discretization with 100 points in a range of characteristic lengths in the range of 10 nm to 100 pm. The population balance equation was implemented in the volume-based incarnation. The solubility (modelled according to the modified Apelblat equation, Apelbalt andManzurola., J. Chem. Thermodyn. 1527-1533, 1997) was fitted on experimental data on (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide and was in this example 0.0625 g / gsoiution at 20°C and at a solvent composition 80 / 20 w / w% Acetone / Water. It had been experimentally determined that the level of maximum attainable supersaturation before spontaneous nucleation occurs was about 1.5 in the temperature range of 30 to 50 °C for (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide. The initial concentration of the API (corresponding to the concentration obtained upon mixing the streams, as in Example 1) was set to 0.044 g / gsoiution. The seed population used for the simulations followed a lognormal functional form, with the tenth, fiftieth, and ninetieth quantiles of the cube weighted distribution at 35, 80, and 240 nm, respectively. The crystallisation started at 50°C, where the API was completely dissolved, cooled down to 35°C, where a supersaturation ~1.4 was reached and where the seeding material was introduced. After 30 minutes at such temperature, the simulations continued by cooling the system further to 10°C (with a simulated time of 8 hours), followed by another hour at 10°C (see Figure 8 below for the temperature profile). The simulations were performed taking size-dependent solubility into account. The simulations were repeated with the same set-up, changing only the amount of seeds, with relation to the total dissolved API, yielding a different final particles size distribution. The results are reported in the table 1 below.

[0327] Table 1 BHC243022-FC

[0328] -63-

[0329] Table 1 shows that the average d90 Particle size in pm calculated from d90 particle size distribution increased between 1 wt% and 0.00 lwt% seeding material if less seeding material was employed. Between 0.00 lwt% and 0.000 lwt% this trend was inverted and then occurred again between 0.0001wt% and 0.000000 lwt% .

[0330] Example 3 Asymmetric seeding material

[0331] Example 3a Modelling

[0332] To model the case that the used particles were geometrically asymmetric in that one of length, width or height was not of nanometer size a standard population balance equation (see for instance D. Ramkrishna, Population Balances, Theory and Applications to Particulate Systems in Engineering, 2000), with one characteristic length was used. The particles were described in the model by a simplified geometrical shape, namely as a square-based prism, whose height was bigger than its width. The geometric figure used to model this particular crystal morphology is also typically referred to as “needle-like” or simply “needle” in the crystallization literature. Using the aforementioned geometry, the crystals volume and surface can be computed as function of the characteristic length times a volume shape factor (kv = 0.112) and a surface shape factor (ks = 1.556), respectively. In this exemplary simulation the length of the needles or the height of the prisms was used and had a d90 value in range of 1 pm - 2pm. The other dimensions were calculated from the aspect ratio and had an d90 value in range of 10-700 nm. The values for the shape factors have been derived assuming for all particles a constant Aspect Ratio (AR, defined as the ratio between the needle length and the needle width) value of 3. The population balance equation was coupled with the standard mass balance on the active ingredient, relating the crystal mass via the second moment to the free solute to account for the dynamic variation of supersaturation. The high- resolution finite volume method (with Koren flux limiter) was implemented in Python, according to the method reported by Zhang et al. (Comp & Chem Eng., 108612-183, 2024), and a nonlinear grid (log-scaled) was chosen to simulate the system, providing a discretization with 100 points in a range of characteristic lengths in the range of 10 nm to 100 pm. The population balance equation BHC243022-FC was implemented in the volume-based incarnation. The solubility (modelled according to the modified Apelblat equation, Apelbalt et al., J. Chem. Thermodyn. 1997) was fitted on experimental data on (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide and was in this example 0.01 g / gsoiution. The initial concentration of the API (corresponding to the concentration obtained upon mixing the streams, as in Example 1) was set to 0.044 g / gsoiution. The seed population used for the simulations followed a lognormal functional form, with the tenth, fiftieth, and ninetieth quantiles of the cube weighted distribution at 35 , 80, and 240 nm, respectively. The simulation was performed assuming a supersaturation level of- 4.3 (i.e. this was the calculated maximum attainable supersaturation level) was instantaneously attainable and that the system was isothermal. The total simulation time was set to 1 hour. The set of kinetic parameters for primary nucleation and linear crystal growth were kept unchanged in both cases. The initial seed population for all simulations was identical . The simulation took size-dependent solubility into account. The results are shown in Table 2.

[0333] Table 2

[0334] Table 2 shows that the average d90 Particle size in pm calculated from d90 particle size distribution increased between 1 wt% and 0.01wt% seeding material if less seeding material was employed. Between 0.01wt% and 0.001wt% this trend was inverted and then occurred again between 0.001wt% and 0.000000 lwt% .

[0335] When comparing the values from Table 1 with those of Table 2 it can be seen that the method described herein, and modelled in Example 3a, resulted in smaller average particle sizes i.e. the amount of nanoparticles as seeding material can be reduced to still generate the same average BHC243022-FC

[0336] -65- particle size as with the method modelled in Example 2. Moreover, it can be seen that the model of Example 3a can be used to predict which amount of seeding material will results in which average particle size. Hence a person skilled in the art can model which amount of seeding material is required for a desired particle size. In this respect it should be noted that also the values for nanoparticles which are characterized by a nano-particle size in all of length, diameter or height of a d90 in the range of 10 nm to 999 nm can be used in the model as described in this Example 3a instead of nanoparticles which are characterized by a size of a d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS) in at least one of length, diameter or height as well as a size of a d90 in the range of 1pm to 2pm as measured by Dynamic Light Scattering (DLS) in at least one of length, diameter or height as measured by Dynamic Light Scattering (DLS). In addition the model as described in this example 3a can be used to determine the values for nanoparticles which are of non-needle like appearance.

[0337] The simulations of Example 2 and Example 3 a were also carried out without taking size dependent solubility into account (data not shown). It was found that if size dependent solubility was not accounted for the modelled resulting particle sizes predicted that the particle size generally increased if less nanoparticles acting as seeding material were present but these modelled resulting particle sizes did not correspond well with the experimentally determined particle sizes of Examples lb and 3b.

[0338] Example 3b Experimental verification of the simulation employing asymmetric seeding material

[0339] In order to show that the method described herein also works using asymmetric seeding material the following experiment was carried out. The seed amount was calculated with respect to the proportion of (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino) propanamide contained in the supersaturated solution. For the crystallization experiment 100g of a homogenous supersaturated (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide solution consisting of 80 / 20 % w / w Acetone / Water with dissolved 8.7 g of (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide and an aqueous nanosuspension comprising (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4- fluoro-anilino)propanamide nanoparticles as seeding material at 0.1 %, 0.05 %, 0.01%, 10E-5 %, 10E-7%,10E-8%, 10E-9%,w.r.t (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro- anilino)propanamide were provided. The weight percentage (wt) values are given with respect to the total amount in the final solution during crystallisation i.e. with respect to the total concentration of substrate i.e. substance e.g. active ingredient in the aqueous nanosuspension and the BHC243022-FC

[0340] -66- homogeneous saturated solution combined. All aliquots of nanoseeds were obtained by dilution of a 10%wt suspension previously prepared as described in example 1 but with the respective amount of (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide to achieve a given %wt.

[0341] The solution and the suspension were then drained from the storage vessels at room temperature at the same flow rate (20 mL / minute ) and thereby gave a ratio of 1 : 1 when encountering each other in the T-shaped mixer. The T mixer in this experiment was characterized by 1mm internal diameter, 1.5 x 1.5 cm horizontal and vertical T-sides and was made from Teflon. It was connected to the storage vessels and the end vessel via Teflon tubes of 50 cm length and of 1.6 mm internal diameter. All vessels referred to for the experiments were Schott bottles of 500 mL size. Upon mixing the solution and the suspension at the above specified 1:1 ratio supersaturation was generated and crystallization was initiated. The crystals produced in the tubes, along with the desupersaturating solution, were flowed to the end-vessel, from which they were sampled with a pipette to perform microscope (Keyence Digital Microscope VHX-7000) and LD analysis (Malvern 3000). The analysis under microscope occurred immediately upon depositing the suspension on a laboratory glass slide. The analysis in DL occurred upon redispersing filtered crystal in saturated water and adding a small aliquot of Tween80. Once the experiment was completed, the crystals were filtered under vacuum, in a standard glass funnel filter, with a standard lab paper filter (45 mm diameter, Schleicher & Schuell) and washed with water. Preliminary tests with LD and microscope indicated that filtration and washing did not change the size of the crystals. Each analysis was repeated both at the end of the dosing time and 1 hour after the dosing was completed, to verify for crystal growth. During the holding time, the crystals were stirred at 200 rpm with a magnetic stirrer.

[0342] The results corresponded to the results of Table 2. Hence it is possible to adjust the size of the generated microparticles via the amount of nanoparticles in the aqueous nanosuspension. In addition the results demonstrate that it was sufficient if the nanoparticles comprised in the aqueous solution were characterized in that at least one of their length or width or height was in the range of 10 nm to 999 nm while at least one of their length or width, or height was in the range of 1pm to 2pm for the method to work.

Claims

BHC243022-FC-67-CLAIMS1. A method for generating micro-particles comprising the steps of a) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of the active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the same active ingredient in the range of 2 to 20, d) optionally isolating micro-particles.

2. The method of claim 1, wherein the active ingredient is (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide and the aqueous nanosuspension comprising nanoparticles as seeding material comprises a concentration in the range of 0.99 % wt to 0.001% wt nanoparticles as seeding material.

3. The method of claim 1 comprising the steps of a) Providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of active ingredientin the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of the same active ingredient in a crystallization medium wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and DioxaneBHC243022-FC-68- c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the active ingredient in the range of 2 to 20, d) optionally isolating micro-particles.

4. The method of claim 1 comprising the steps ofProviding an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of the active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined providing a homogenous saturated solution of the active ingredient in a crystallization medium wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the substance in the range of 3 to 10, d) optionally isolating micro-particles.

5. The method of claim 1 comprising the steps of a) Providing an aqueous nanosuspension comprising (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), laser diffraction or electron microscopy and at least one stabilizer, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of (R)-2-(N-[4-amino- 5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in the aqueous nanosuspension and the homogeneous saturated solution combined b) providing a homogenous saturated solution of (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in a crystallization medium wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and DioxaneBHC243022-FC-69- c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising nanoparticles as seeding material at a supersaturation level with respect to the (R)-2-(N-[4- amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in the range of 3 to 10, d) optionally isolating micro-particles.

6. The method of claim 5, wherein the concentration of the seeding material is in the range of 0.1% wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) K12 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0. lwt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1:10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2: 1 to 1:

2. wherein the active ingredient is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4- fluoro-anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane and wherein the supersaturation level with respect to (R)-2-(N-[4-amino-5-(4- methoxybenzoyl)thiazol-2-yl]-4-fluoro-anilino)propanamide in the range of 3 to 10.

7. The method of claim 1 comprising the steps of a) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), wherein the concentration of the seeding material is in the range of 0.5 % wt to 0.001% wt, where the % wt is calculated with respect to the total concentration of active ingredient in the aqueous nanosuspension and the homogeneous saturated solution combined and at least one stabilizer b) providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material wherein the mixing starts in mixing element and said mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles asBHC243022-FC-70- seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time, d) optionally isolating micro-particles wherein in step c) the supersaturation level with respect to the active ingredient is in the range of 2 to 208. Method according to claim 7 comprising the steps of a) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer b) providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time, d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the active ingredient is in the range of 3 to 5 wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) KI 2 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0.1 wt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1: 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2:1 to 1:

2. wherein the active ingredient is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4- fluoro-anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane.

9. Method according to claim 7 comprising the steps ofBHC243022-FC-71- c) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizer d) providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y -shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time in a ratio of the flow rates of homogenous saturated solution to aqueous nanosuspension comprising nanoparticles as seeding material in the range of 2: 1 to 1:2 d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the active ingredient is in the range of 3 to 5 wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) K12 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0. lwt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1: 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2: 1 to 1:

2. wherein the active ingredient is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4- fluoro-anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane.

10. Method according to claim 7 comprising the steps of a) providing an aqueous nanosuspension comprising active ingredient nanoparticles as seeding material characterized by a nano-particle size in at least one of length, diameter or height of a volume based d90 in the range of 10 nm to 999 nm as measured by Dynamic Light Scattering (DLS), and at least one stabilizerBHC243022-FC-72- b) providing a homogenous saturated solution of the same active ingredient in a crystallization medium c) mixing the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material wherein the mixing starts in at least one mixing element and said at least one mixing element is a T- or Y-shaped tubing and the homogenous saturated solution and the aqueous nanosuspension comprising active ingredient nanoparticles as seeding material are provided to the at least one mixing element as at least one continuous fluid stream each at the same time in a ratio of the flow rates of the homogenous saturated solution to aqueous nanosuspension comprising active ingredient nanoparticles as seeding material of 1 : 1 , d) optionally isolating micro-particles. wherein in step c) the supersaturation level with respect to the active ingredient is in the range of 3 to 5, wherein the concentration of the seeding material is in the range of 0.1 % wt to 0.001% wt, wherein the aqueous nanosuspension comprises HPC in the range of 2wt% to 3wt%, polyvinylpyrrolidone (PVP) KI 2 in the range of 0.01 wt% to 10 wt% and SDS in the range of 0.1 wt% to 0.3 wt%, wherein PVP is present in an amount of PVP to HPC in the range of 10: 1 to 1: 10 , wherein the ratio of HPC and PVP together to the nanoparticles is in the range of 2:1 to 1:

2. wherein the active ingredient is (R)-2-(N-[4-amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4- fluoro-anilino)propanamide wherein the crystallization medium is selected from the group consisting of acetic acid, ethanol, methanol, DMSO and Dioxane.

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