The present invention relates to a new solid state of Tiotropium Bromide, a process for preparing it and its use as medicament and as active ingredient in a pharmaceutical composition for the treatment of respiratory complaints, particularly for the treatment of chronic obstructive pulmonary disease (COPD), bronchitis, emphysema and asthma.

BACKGROUND TO THE INVENTION

Tiotropium bromide (compound identified by CAS registry number 136310-93-5) was described for the first time in 1991 by Boheringer Inghelheim (EP 0418716) and presents the following structural formula:

Tiotropium bromide is an anticholinergic bronchodilator with a long-lasting effect, 24 hours, which may be used to treat respiratory complaints, particularly COPD (chronic obstructive pulmonary disease), bronchitis, emphisema and asthma.

Tiotropium bromide is preferably administered by inhalation: suitable inhalable powders packed into appropriate capsules may be used (Spiriva ^® ; US7694676 and US8022082) or alternatively, it may be administered by the use of inhalable aerosols (EP2201934).

The correct manufacture of the above mentioned compositions, suitable for the administration of a pharmaceutically active substance by inhalation, is based on the definition of physical parameters, like a particular crystalline form (see for example US6608055 and US677423) and a defined particle size distribution (US7070800), which are connected with the nature of the active substance itself. From literature data, Tiotropium Bromide is described to exist in different polymorphic forms as well as in an amorphous form. In more details, Tiotropium Bromide is described to exist in a crystalline monohydrate form (US6777423), in several anhydrous (US6608055, WO2006/1 17300, EP1682542) and solvate (US7879871 , W02010/101538, WO201 1/015882) polymorphic forms. Some of these polymorphic forms are unstable and may change. For example, the monohydrate form described in US6777423 may be easily transformed after a mild heating at 40°C for few hours into the corresponding anhydrous form described in US6608055.

Since the quality of a pharmaceutical formulation requires that an active substance should always have the same crystalline modification, the stability and properties of the crystalline active substance are subject to stringent regulatory requirements. There is the need of a crystalline form of tiotropium bromide which is stable to humidity and to mechanical treatments, like the micronization or other milling techniques, and which meets the high demands mentioned above for any pharmaceutically active substance.

SUMMARY OF THE INVENTION It has now surprisingly been found that a crystal modification of tiotropium bromide meeting the above requirements can be obtained in the form of a cocrystal with lactose. Unexpectedly, this cocrystal is stable towards the influence of moisture and humidity and to physical treatments like the micronization.

In the novel cocrystal the components tiotropium bromide and lactose are present in an almost stoichiometric ratio, as determined for example by NMR spectroscopy. Therefore, the present invention relates to a tiotropium bromide-lactose cocrystal in which the components tiotropium bromide and lactose are present, with the limit of resolution of the employed analytical technique (ie H-NMR), in a ratio of about 1 : 1.

This novel cocrystal is characterized by a single endothermic event at about 191-3°C determined by DSC and by an X-Ray spectrum with characteristic 2theta values at 13.08; 14.16; 14.68; 17.90; 18.58; 19.06; 19.44; 21.02; 22.58; 23.24; 25.26; 26.20; 27.24; 28.08; 28.42; 29.96; 30.18; 31.80; 34.50; 34.82; 35.58; 38.70; 39.26; 41.52 and 50.06.

The present invention also relates to the cocrystal herein disclosed for use as a medicament, in particular for the treatment of respiratory complaints, particularly for the treatment of COPD and/or asthma.

The present invention also relates to a pharmaceutical composition comprising the above cocrystal as active ingredient.

The present invention also relates to a process for the preparation of the crystalline tiotropium bromide forms according to the present invention.

With a view to administration by inhalation, it is essential to provide the active substance in finely divided form. For this purpose, the cocrystal according to the invention is obtained in finely divided form using methods known in the prior art. Methods of micronising active substances, like the air jet mill or the conical screen mill techniques, are known in the art. Preferably, after micronizing, the active substance has a mean particle size of 0.5 to 10 μιη, preferably 1 to 6 μηη, more preferably 1.5 to 5 μιτη.

Tiotropium bromide cocrystal according to the invention is particularly well suited to the preparation of, for example, pharmaceutical formulations for administration by inhalation such as inhalable powders or for example propellant-containing aerosol formulations, particularly inhalable powders and propellant- containing aerosol suspensions. These pharmaceutical formulations or compositions may contain in addition to the crystalline Tiotropium cocrystal according to the invention one or more additional active ingredients selected from the group consisting of betamimetics, EGFR inhibitors, PDEIV-inhibitors, steroids, and LTD4 antagonists, optionally together with a pharmaceutically acceptable excipients. It is intended that also other active ingredients can be added according to the general common knowledge.

The present invention will be now illustrated in detail, also by means of Figures and Examples.

DESCRIPTION OF THE FIGURES Figure 1 shows a representative DRX spectrum of Tiotropium Bromide cocrystal with lactose (third DRX spectrum on the bottom) overlapped with the DRX spectra of the employed starting materials (from the top respectively the DRX spectra of lactose monohydrate and of Tiotropium Bromide).

Figure 2 shows the DSC-thermograms of the lactose cocrystal with Tiotropium bromide with an endothermic event at about 191-193°C. Figure 3 shows H-NMR spectra (500 MHz) were recorded of the obtained cocrystal. Figure 3A shows expanded zone of the same sample from 0 to 5 ppm.

Figures 4 and 4A show the DRX spectra of a Tiotropium Bromide monohydrate before and after the heating at 40°C at 50% UR for 72 hours.

Figure 5 shows DRX data of Tiotropium Bromide cocrystal with lactose micronized using air jet mill technique.

DETAILED DESCRIPTION OF THE INVENTION

Within the frame of the present invention, cocrystal are solids that are crystalline materials composed of two or more molecules in the same crystal lattice where each component is defined as either an atom, ion, or molecule (Stahly, G. P. (2009). "A Survey of Cocrystals Reported Prior to 2000". Crystal Growth & Design 9 (10): 4212; Regulatory Classification of Pharmaceutical Co-Crystals Edited by U.S. Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER) April 2013). Cocrystal exist in their neutral states and interact via nonionic interactions, as opposed to an ionic interaction, which would classify this crystalline solid as a salt form.

According to the present invention, the process for the preparation of Tiotropium Bromide lactose cocrystal comprises the following steps: a) mixing Tiotropium Bromide and lactose in a relative molar ratio comprised between 1.0 and 1.3 in dimethylsulfoxide to a final concentration comprised between 1 and 14 M under stirring at room temperature to provide a suspension;

b) heating said suspension to obtain a solution;

c) adding portion wise under stirring to said solution, an aprotic organic solvent; preferably in a relative ratio, with respect to the dimethylsulfoxide, comprised between 7 and 9 times in volume;

d) cooling the reaction mixture under stirring to obtain a precipitate;

e) recovering said precipitate; f) drying said precipitate under vacuum;

g) optionally the dry product can be micronized

In another embodiment of the present invention, the process for the preparation of the cocrystal herein disclosed further comprises after step e) and before step f) the following steps: e') dispersing said precipitate from step e) in acetone to give a dispersion, and e") recovering said precipitate from said dispersion and washing it with acetone, preferably about 5 volumes respect to the volume of employed dimethylsulfoxide.

The employed lactose in step a) can be in crystalline form monohydrate, anhydrous or amorphous, preferably in the monohydrate alpha crystalline form Preferably the temperature of heating said suspension in step b) is comprise in a range of 50-55°C

Preferably the aprotic organic solvent in step c) is selected among organic solvents with a log P value comprised between -0.24 and 1.14, like for example acetone, methylethylketone, ethylacetate, methyl acetate, n-propylacetate, more preferably acetone and ethyl acetate. Portion wise addition can be made according to normal experience, for example in a time ranging between few minutes and 2 hours, depending also on the operating conditions. A convenient timing is about 30'. This step is preferably carried out at 50-55°C

Preferably the cooling rate in step d) is of about 1-2°C minute. The preferred temperature is of 20-25°C and can be maintained for the desired time, typically for a time interval ranging from 0.5 to 5 hours, preferably from 1 to 4 hours, more preferably from about 2 The steps e'), e")and g) can be performed optionally since they do not influence the solid state of the obtained crystalline form but only the chemical purity; alternatively the wet product obtained at point e) can be directly treated according to step f).

Recovery of the precipitate can be done with any conventional means, filtration being the preferred one.

The temperature range of step f) can be increased up to 60°C without affecting the solid state of the obtained crystalline form. Typically, drying temperature ranges around 20-40°C. Drying time can be selected according to normal experience, for example more than 24, up to 96 hours, for example from 24 to 72 hours.

The step g) can be realized using conventional milling techniques like the Air Jet Mill technique or Conical screen mill without changes in the solid state; the preferred characteristic of the micronized product include but it is not limited to particle size distribution with a D90 <10 μιη.

The cocrystal according to the present invention, in particular the cocrystal obtained by the process herein disclosed, is characterized by a single endothermic event at about 191-3°C determined by DSC and by an X-Ray spectrum with characteristic 2theta values at 13.08; 14.16; 14.68; 17.90; 18.58; 19.06; 19.44; 21.02; 22.58; 23.24; 25.26; 26.20; 27.24; 28.08; 28.42; 29.96; 30.18; 31.80; 34.50; 34.82; 35.58; 38.70; 39.26; 41.52 and 50.06.

The stoichiometry of the components of the cocrystal of the present invention is about 1 : 1. The determination of the stoichiometry can vary around the ration of 1 : 1 depending on the analytical technique used. The ratio of the components can vary around the ratio of 1 :1 on condition that the stability of the cocrystal is not affected.

According to another object of the present invention, the cocrystal herein disclosed is for use as a medicament, in particular for the treatment of a respiratory complaint, more in particular chronic obstructive pulmonary disease (COPD), asthma, bronchitis and emphysema. The dosage, way of administration and clinical indication can be decided by the expert of the art, based on the general knowledge, for example as shown in EP 0418716 for aerosol and tablets. See also US 200211529, US 20030087927 and EP 2201934 for aerosol application.

Another object of the present invention is a pharmaceutical composition comprising the above cocrystal as active ingredient. Pharmaceutical compositions according to the present invention are characterized by the presence of the cocrystal herein disclosed. Said compositions comprise any of conventional vehicles, excipients, formulative ingredients and can be prepared according to the general knowledge in this art. A general reference can be found for example in Remington "The Science and Practice of Pharmacy, 21 ^st edition Pharmaceutical Press. In a preferred embodiment, said composition is suitable for administration by inhalation. In a more preferred embodiment, in said composition for inhalation, said cocrystal has a mean particle size of 0.5 to 10 μηη, preferably 1 to 6 μιη, more preferably 1.5 to 5 μηι.

Pharmaceutical compositions for inhalation, in particular for aerosol inhalation are well known in the art and do not need any specific description for the present invention, the general common knowledge being sufficient, see for example Pharmaceutical Inhalation Aerosol Technology, Anthony J. Hickley ed., Marcel Dekker, Inc. 2004.

Embodiments of the pharmaceutical compositions of the present invention comprise those compositions disclosed in US7694676 and US8022082, wherein the active ingredient is the cocrystal of the present invention.

The following Examples further illustrate the present invention. MATERIALS AND METHODS

¹ H NMR analyses were performed at 500 MHz with a Bruker FT-NMR AVANCE™ DRX500 spectrometer. Infrared spectra (IR) were registered on a Perkin Elmer instrument (Mod FTIR Spectrum one) equipped with universal ATR sampling. DSC were registered on a Perkin Elmer instrument (Mod. DSC7) at a heating rate of 10 °C/min from 40°C to 260°C. Between 3 and 6 mg of sample were used and all samples were crimped in hermetically sealed aluminum pans X-Ray Diffraction spectra were registered by means of diffractometer (Rigaku-D-Max) from a start angle [1/2 2-theta] of 5.000 to 60.000. The diffraction diagrams were obtained employing a Cu anode (Ka = 1 ,54060 A and Ka = 1 ,54439 A). The HPLC method utilized to check the quality of Tiotropium Bromide in the examined samples is the same as described in the European Pharmacopoeia 7.0. Example 1

Preparation of Tiotropium Bromide Cocrystal with lactose in dimethylsulfoxide and acetone

Tiotropium Bromide (4.25g; 8.99 mmoles) and lactose monohydrate (3.58g; 9.9 mmoles) were dispersed at room temperature in dimethylsulfoxide (7.2 ml). The mixture was heated under stirring at the temperature of 50-55°C to obtain a limpid solution. Then acetone (55 ml) was added dropwise in 30' maintaining the reaction mixture under stirring at 50-55°C. The obtained solution was cooled down at 20- 25°C and kept at this temperature under stirring for 2 hours. After this period a suspension was obtained. The precipitate was recovered by suction and the wet product slurried in acetone (14.9 ml) under stirring for 90'. The product was recovered by filtration, washed on the filter with acetone (4 times with 8.5 ml each) and dried under vacuum at 40°C for 48 hours to afford 6.76g (8.1 1 mmoles; 90% molar yields) of dry product.

The obtained crystals were analysed by X-Ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and H-NMR (500 MHz) indicating that a new crystalline form, namely a cocrystal of Tiotropiumbromide with lactose is formed.

A representative DRX spectrum of Tiotropium Bromide cocrystal with lactose is shown in Figure 1 (third DRX spectrum from the bottom) overlapped with the DRX spectra of the employed starting materials (from the top respectively the DRX spectra of lactose monohydrate and of Tiotropium Bromide). The list of the characteristic diffraction peaks including normalised intensities of Tiotropium Bromide cocrystal is shown in table 1.

Table 1

2theta Intensity l/lo

11.360 420 21

12.340 453 23

13.080 366 18

13.520 843 42

14.160 1026 51

14.680 1098 54

15.280 659 33 theta Intensity l/lo5.960 696 356.300 524 266.960 872 437.900 1472 738.580 1897 939.060 1847 919.440 1938 950.460 791 391.020 826 411.720 819 412.580 1712 843.240 2044 1003.980 919 454.380 853 425.260 1499 746.200 1156 577.240 1240 618.080 1554 778.420 1935 959.960 1043 520.180 1024 510.500 819 410.900 924 461.800 1399 692.340 960 472.980 839 423.440 937 46 theta Intensity l/lo4.500 1032 514.820 1424 705.580 1196 596.380 899 446.880 964 487.220 917 457.740 913 457.960 982 498.700 1163 579.260 1035 519.880 833 411.520 1396 691.940 976 483.400 1005 503.600 989 494.900 1074 535.360 893 446.920 1001 497.480 932 468.060 958 479.540 1107 550.060 1023 511.820 995 494.440 990 495.220 946 476.340 960 478.100 926 46 The DSC-thermograms of the lactose cocrystal with Tiotropium bromide shows an endothermic event at ca. 191-193°C indicating melting of this material. The obtained DSC-diagram is depicted in Figure 2.

In order to get an idea on the stoichiometry of the obtained cocrystal H-NMR spectra (500 MHz) were recorded. The samples were dissolved in d6-DMSO for analysis. The corresponding spectrum is shown in Figures 3 and 3A. In addition to the characteristic H-NMR signals of tiotropium there is a signal at 4,17 and 4, 18 ppm which is indicative of H1 ' of lactose (integration 1 H; ref Hyunnsook Ko et al. Bull. Korean Chem. Soc. 2005, Vol 26, No12, 2001-6). Integration of this signal compared to the signal at 4,12 ppm of CH 1 and 5 (integration 2H; ref. Zhenguang Lin et al. Spectrochimica Acta Part A 75 (2010), 1 159-1162) of Tiotropium shows that the cocrystal has a stoichiometry which is close to 1 : 1. Example 2

Preparation of Tiotropium Bromide Cocrystal with lactose in dimethylsulfoxide and ethyl acetate

Tiotropium Bromide (2.0; 4.23 mmoles) and lactose monohydrate (1.68; 4.66 mmoles) were dispersed at room temperature in dimethylsulfoxide (3.4 ml). The mixture was heated under stirring at the temperature of 50-55°C to obtain a limpid solution. Then ethylacatete (48 ml) was added dropwise in 30' maintaining the reaction mixture under stirring at 50-55°C. The obtained solution was cooled down at 20-25°C and kept at this temperature under stirring for 2 hours. After this period the product was recovered by filtration, washed on the filter with ethyl acetate (4 times with 5 ml each) and dried under vacuum at 20°C for 48 hours to afford 2.83g (3.39 mmoles; 80% molar yields) of dry product.

The physico-chemical data of the obtained solid are the same of the product isolated in the Example 1. Stability data of the crystalline form of Tiotropium Bromide Cocrystal with lactose

Tiotropium Bromide Cocrystal obtained according to the Example 1 was analyzed by DRX (powder) at 1 , 2 and 3 months in the following storage conditions in order to verify the stability of this crystalline form. The results are shown in Table 2.

Table 2

In the same storage conditions, and particularly using as primary packaging polyethylene bag, Tiotropium Bromide may absorb or loose water, thus changing the crystalline form. In fact, this is an equilibrium that is driven by the relative humidity of the environment. This change in the solid state can be nearly quantitative: so for example the hydrate form described in US6777423 changes into the corresponding anhydrous form described in US6608055 after heating at 40°C at 50% UR for 72 hours. See the enclosed DRX spectra of a Tiotropium Bromide monohydrate before and after the above described treatment (Figures 4 and 4A).

Micronization of Tiotropium Bromide Cocrystal with lactose

A sample of Tiotropium Bromide Cocrystal obtained according to Example 1 was micronized using the air jet mill technique in order to verify if this physical treatment affected the quality and the solid state of this product. The obtained analytical results (particle size distribution and chemical purity determined by HPLC) are shown in Table 3; the DRX data of the micronized material confirm that the solid state is not changed after micronization (Figure 5).

Table 3

HPLC purity of

D10 D50 D90 Titropioum

Bromide

Unmilled Cocrystal 2.03 microns 16.6 microns 178 microns 99.91 %

Milled Cocrystal 0.67 microns 1.95 microns 5 microns 99.88%