HK1140479B - Mono-hydrochloric salts of an inhibitor of histone deacetylase - Google Patents

Description

Monohydrochloride salts of histone deacetylase inhibitors

Technical Field

The present invention relates to novel crystalline forms of mono-HCl salts and mono-HCl salt hydrates of JNJ-26481585, an inhibitor of histone deacetylase. The invention also relates to processes for the manufacture of these forms, to intermediates used in these processes, to pharmaceutical compositions comprising these forms and to the use of these forms for medical treatment, for example as a medicament for inhibiting proliferative conditions such as cancer or leukaemia.

Background

Many solid pharmaceutical entities may exist in different physical forms, e.g. in amorphous form, in one or several crystalline forms (e.g. anhydrous or solvate form), in the form of a mixture of different crystalline forms or as a mixture of amorphous and crystalline forms.

The amorphous form is a form in which no three-dimensional long-range ordered arrangement exists. In amorphous form, the position of the molecules relative to each other is substantially random, i.e., an irregular arrangement of molecules on a lattice structure. Amorphous and disordered materials generally have improved properties, but creating and stabilizing this state can be a great challenge.

A crystalline or crystalline form is one in which the positions of the molecules relative to each other are organized according to a three-dimensional lattice structure. Crystalline forms generally include polymorphs and pseudopolymorphs (pseudopolymorphs). Polymorphs are different crystalline forms of the same compound in the solid state due to different molecular arrangements. Different polymorphs have different crystal structures due to the different assembly of molecules in the crystal lattice. This results in different crystal symmetries and/or unit cell parameters. Polymorphs differ from each other by their physicochemical parameters rather than their chemical composition. Polymorphism is often difficult to control and challenging for medical galenicals (galentists). Pseudopolymorphism, also known as solvate, is a particular condition of solid-state crystalline forms in which stoichiometric or non-stoichiometric amounts of solvent molecules are present or incorporated into the lattice structure of the compound. The water solvate is also referred to as a hydrate.

Solid state chemistry is of interest to the pharmaceutical industry, particularly those involved in the development of suitable dosage forms. For example, solid state deformation can severely affect the stability (shelf life) of a drug. Metastable pharmaceutical solid forms may be transformed into crystalline structures (e.g., from amorphous to crystalline) or solvates/desolvates in response to environmental conditions, processing, or time changes.

Different crystalline or amorphous forms of the same drug may have significant differences in such pharmaceutically important properties as dissolution rate, thermodynamic solubility, and bioavailability. The rate of dissolution of the active ingredient in the gastric juices of a patient may be of therapeutic importance as it allows an upper limit to the rate at which an orally administered active ingredient can reach the patient's blood circulation. The rate of dissolution is therefore a consideration when formulating solid dosage forms and liquid pharmaceutical products (e.g., syrups or elixirs).

Likewise, different crystalline or amorphous forms may have different processing properties, such as hygroscopicity, flowability, compactability, etc., which may affect their suitability as commercially produced active pharmaceuticals.

In the clinical development of medicine, if the polymorphic form is not held constant, the precise dosage forms used or studied in each batch may not be comparable. Methods for manufacturing compounds having a high purity of a selected polymorphic form are also desirable when the compounds are used in clinical studies or commercial products, as the presence of impurities may produce undesirable toxicological effects. Certain polymorphic forms may exhibit enhanced thermodynamic stability or may be manufactured in high purity and more rapidly in large quantities and are therefore more suitable for inclusion in pharmaceutical formulations.

JNJ-26481585 has the following structure:

the compounds are inhibitors of Histone Deacetylase (HDAC).

WO 2006/010750 published on 2.2.2006 discloses JNJ-26481585.C₂HF₃O₂Amorphous forms of the salts and di-HCl salts and methods for making them.

JNJ-26481585.C, originally described in WO 97/21701₂HF₃O₂The synthesis of the salts is shown in scheme 1.

Wherein, in step 1, the intermediate of formula (III) is prepared by reacting the intermediate of formula (I) with formaldehyde of formula (II) in methanol in the presence of sodium tetrahydroborate.

In step 2, the intermediate of formula (IV) is prepared by reacting the intermediate of formula (III) with sodium hydroxide in ethanol.

In step 3, the intermediate of formula (V) is prepared by reacting the intermediate of formula (IV) with O- (tetrahydro-2H-pyran-2-yl) -hydroxylamine in the presence of a suitable reagent such as N' - (ethylcarbonylimino) -N, N-dimethyl-1, 3-propanediamine, monohydrochloride (EDC) and 1-hydroxy-1H-benzotriazole (HOBT). The reaction is carried out in a mixture of dichloromethane and tetrahydrofuran.

In step 4, hydroxamic acid C of formula (VI)₂HF₃O₂Salts are prepared by reacting an intermediate of formula (V) with trifluoroacetic acid. The reaction was carried out in methanol.

Scheme I

Alternatively, as initially described in WO 97/21701, JNJ-26481585.2HCl salt was prepared by reacting the intermediate of formula (III) with hydroxylamine in the presence of sodium hydroxide. The reaction is carried out in methanol and the further conversion to the di-HCl salt is prepared in ethanol.

The process disclosed in WO 2006/010750 is not suitable for mass production as a result of low yields and high impurity levels in the different processing steps, followed by several chromatographic steps. The use of chromatography to purify compounds is expensive and environmentally unfriendly due to the consumption of solvents and specialized equipment required for large scale chromatography.

The problem addressed by the present invention is to provide novel crystalline forms of the mono-HCl salt and mono-HCl salt hydrate of JNJ-26481585. Another aspect of the invention is a process for obtaining the novel crystalline mono-HCl salt and mono-HCl salt hydrate forms in high yield and purity. Advantageous properties of the HCl form of the invention are advantageous physicochemical properties, including its non-hygroscopic nature and chemical stability contributing to the drugability of this compound.

Drawings

Fig. 1 is a graphical representation of an Infrared (IR) spectrum of form I.

Figure 2 is a graphical representation of X-ray powder diffraction (XPRD) of form I.

Fig. 3 is a Differential Scanning Calorimetry (DSC) curve of form I.

FIG. 4 is the weight change of form I as a function of relative humidity.

FIG. 5 is a graphical representation of the adsorption-desorption (ADS/DES) curve for form I.

Fig. 6 is a graphical representation of the IR spectrum of form II.

Figure 7 is a XPRD representation of form II.

Figure 8 is a DSC curve for form II.

Figure 9 is the change in form II weight as a function of relative humidity.

FIG. 10 is a graphical representation of the ADS/DES curve for form II.

Fig. 11 is a graphical representation of the IR spectrum of hydrate form I.

Figure 12 is a XPRD representation of the hydrate form.

FIG. 13 is a stack of XPRD spectra for slurry conversion studies of form I and form II in ethanol at different temperatures.

Figure 14 is a XPRD spectrum overlay of slurry conversion studies of form I and form II in ethanol/water (90/10, vol/vol%) at different temperatures.

Figure 15 is a stack of XPRD patterns from slurry conversion studies of form I and form II in water at different temperatures.

FIG. 16 is a superposition of XPRD spectra for slurry conversion studies of hydrates in ethanol at different temperatures.

Description of the invention

Preparation of intermediates

A. Preparation of intermediates of formula (I)

a) Intermediates of formula (XI) can be prepared by reacting an intermediate of formula (IX) with an intermediate of formula (X) in the presence of a suitable solvent, e.g., a polar or non-polar aprotic hydrocarbon solvent such as toluene, dichloromethane, isopropyl acetate, ethyl acetate, tetrahydrofuran, and the like. Other aromatic or aliphatic aldehydes may be used in the process. The reaction can also be carried out in protic solvents such as methanol, ethanol, isopropanol, and the like. The reaction may be carried out at a temperature of from 25 ℃ to 60 ℃, preferably at a temperature of 45 ℃. The use of higher temperatures is not recommended due to the potential instability of the intermediates of formula (X).

b) Intermediates of formula (VIII) can be prepared by converting an intermediate of formula (VII) in the presence of a suitable oxygenate such as m-chloroperoxybenzoic acid (MCPBA) in a suitable solvent such as a polar or non-polar aprotic hydrocarbon solvent such as toluene, dichloromethane, isopropyl acetate, ethyl acetate, tetrahydrofuran, and the like. The reaction may be carried out at a temperature of-20 ℃ to 40 ℃, preferably at a temperature of 0 ℃ to 5 ℃, more preferably at 0 ℃. At higher temperatures, meta-chloroperoxybenzoic acid is unstable and the intermediate of formula (VIII) may decompose. Complete conversion of intermediate (VII) to intermediate (VIII) can be obtained by adding the appropriate amount of MCPBA. The amount of MCPBA is therefore preferably greater than 1 equivalent.

c) Intermediates of formula (I) can be prepared by reacting an intermediate of formula (VIII) with an intermediate of formula (XI) in the presence of a suitable solvent, e.g., a polar or non-polar aprotic hydrocarbon solvent or mixtures thereof such as toluene, dichloromethane, isopropyl acetate, tetrahydrofuran, diisopropylethylamine or mixtures of other quaternary ammonium bases and ethyl acetate, and the like. The reaction may be carried out at a temperature of-20 ℃ to 40 ℃, preferably at a temperature of 0 ℃ to 5 ℃, more preferably at 0 ℃ to 25 ℃.

This synthesis is accompanied by temporary protection of the aminopiperidine of formula (IX), p-nitrobenzaldehyde of formula (X), formation of an intermediate of formula (XI), allowing more of the substituted ring nitrogen to react preferentially. If this protection is not carried out, a large amount of dimer (A) and isomer (B) is formed as the two nitrogens of the intermediate of formula (IX) react with the intermediate of formula (VIII). Warming the reaction mixture overnight ensured that the intermediate of formula (XI) was fully reacted to the intermediate of formula (I) and any remaining intermediate of formula (IX) was fully converted to the dimer (a), which with the remaining MCPBA could be easily removed in a subsequent acid work-up.

One embodiment of the present invention includes intermediates of formula (XI).

B. Preparation of intermediates of formula (XIII)

a) Intermediates of formula (XII) can be prepared by reacting an intermediate of formula (I) with an intermediate of formula (II) in a suitable solvent. The reaction can be carried out at a temperature of from 50 ℃ to 150 ℃, preferably at a temperature of 110 ℃ (reflux temperature of toluene). The reaction requires azeotropic removal of water. As the solvent, polar or non-polar aprotic hydrocarbon solvents such as toluene, isopropyl acetate and the like can be used. These solvents are well azeotroped with water.

b) The intermediate of formula (XII) is treated with sodium tetrahydroborate in a suitable solvent such as polar or non-polar aprotic and protic hydrocarbon solvents and mixtures thereof, e.g. toluene, isopropyl acetate, ethanol, methanol, isopropanol, and the like. The reduction with sodium tetrahydroborate may be carried out at 0 ℃ to 50 ℃. Preferably at 10 ℃. The temperature during reduction is preferably low to avoid formation of over-reduced impurities.

c) Subsequently, salt formation is carried out with fumaric acid in an acetone/ethanol mixture with 5% v/v water to form the intermediate of formula (XIII).

One embodiment of the present invention includes a fumarate salt of formula (XIII).

C. Preparation of intermediate of formula (XVIII)

In a first attempt to find a better synthetic method for the production of JNJ-26481585, the intermediate of formula (III) was reacted with O- (tetrahydro-2H-pyran-2-yl) -hydroxylamine in the presence of a base and a solvent (but without a coupling agent). The attempt was not successful.

In the second attempt, both the amino and hydroxamic acid groups were protected with acid-labile protecting groups to facilitate simultaneous deprotection-salt formation.

Thus, the intermediate of formula (XIII) is converted to the free base to give an intermediate of formula (III) and further to an intermediate of formula (XIV) wherein R is tert-butyl, benzyl or fulvenyl, followed by hydrolysis with NaOH in ethanol and isolation of the intermediate of formula (XV) by acidification and direct crystallization from the reaction mixture.

Coupling of formula (XV) with O- (tetrahydro-2H-pyran-2-yl) -hydroxylamine under standard amino acid coupling conditions (EDC, HOBT, triethylamine, tetrahydrofuran) gives the intermediate of formula (XVI) in excellent yield. Coupling reactions of intermediates of formula (XV), wherein R is an alkenyl rich group, result in cleavage of some alkenyl rich-groups due to the requirement of triethylamine for optimum coupling.

Deprotection of the intermediate of formula (XVI), wherein R is tert-butyl, is then attempted under various conditions (solvent: ethanol, ethyl acetate, toluene, acetone, methyl isobutyl ketone, dimethylformamide; acid: ethanesulfonic acid, methanesulfonic acid, hydrochloric acid, trifluoroacetic acid). Unfortunately, the only product observed at ambient temperature (2-3 hours) or at 50 ℃ (10 minutes) is the product from cleavage of the indole moiety.

Attempts to hydrogenate the intermediate of formula (XVI), wherein R is benzyl, in the presence of a suitable catalyst (e.g., palladium on carbon) under an atmosphere of hydrogen, were judged unsuccessful due to competitive cleavage (up to 20%) of the N-O bond on the hydroxamic acid product. In another aspect, cleavage of the fulvene group of the intermediate of formula (XVI), wherein R is fulvene group, is successful herein with respect to the intermediate of formula (XVI-a) under mild conditions and by trapping the fulvene group by-product using thiol silica gel. The corresponding free amine of formula (V) gives the compound of formula (XIX) under similar conditions as described below (1.05 equivalents of hydrochloric acid, ethanol, 70 ℃).

a) Finally, neutralization by aqueous sodium hydroxide and extraction in methyltetrahydrofuran gave the free base of intermediate (XIII). The organic layer containing the free base was then subjected to base hydrolysis with 3 molar equivalents of aqueous sodium hydroxide at reflux. The sodium salt in the aqueous layer was then separated from the methyltetrahydrofuran layer and acidified with 5 molar equivalents of HCl at 10 ℃.

The intermediate of formula (XVII) can have a variable water content. Immediately after drying, the water content was 0.7%. When the sample was allowed to stand at atmospheric pressure for 24 hours, the water content increased and stabilized at 8% water, which represents 2 moles of water.

b) The effect of water on intermediate (XVII) is critical, as the following coupling reaction requires a specific amount of water to be successful. The amount of water in intermediate (XVII) is preferably from 15 to 25 v/v%, most preferably about 16 v/v%.

Intermediates of formula (XVIII) can be prepared by reacting an intermediate of formula (XVII) with O- (tetrahydro-2H-pyran-2-yl) -hydroxylamine in the presence of a suitable reagent, such as N- (ethylcarbonylimino) -N, N-dimethyl-1, 3-propanediamine, monohydrochloride (EDC) as coupling reagent. The reaction may be carried out in polar or non-polar aprotic and protic hydrocarbon solvents and mixtures thereof, such as methyltetrahydrofuran, Dimethylformamide (DMF), Dichloromethane (DCM), toluene, isopropanol, ethanol, acetonitrile, ethyl acetate, isopropyl acetate, mixtures thereof and mixtures of one or more different solvents with water, preferably mixtures of ethyl acetate with ethanol, more preferably mixtures of ethyl acetate, ethanol and water. The temperature during the reaction can be between 10 ℃ and 40 ℃, preferably at room temperature.

The reaction is rapid and complete when sufficient water is present. Under dry conditions the reaction is slower, more impurities are present and the reaction rate decreases towards the product.

c) The intermediate of formula (XVIII) may be dissolved in a solvent such as dimethylformamide or dimethylacetamide, preferably dimethylacetamide. The addition of a co-solvent will cause the product to crystallize out. Co-solvents such as acetone, methyl isobutyl ketone or methyl ethyl ketone, preferably methyl isobutyl ketone, may be used. Purification may be carried out at a temperature of 25 ℃ to 90 ℃, preferably 50 ℃ to 70 ℃. The crystallization time should not exceed 5 hours. At higher temperatures or longer crystallization periods, the yield of the final product decreases. Recrystallization may be carried out in a solvent such as ethanol, in the presence of a co-solvent such as methyl ethyl ketone, at a temperature of from 50 ℃ to 70 ℃, preferably at 70 ℃.

One embodiment of the present invention includes the hydrochloride salts of formulas (XVII) and (XVIII).

Preparation of crystalline forms

The intermediate of formula (XVIII) can be converted to the HCl salt of formula (XIX) by the addition of hydrochloric acid in a suitable solvent such as ethanol or methanol, when the reaction mixture is at the desired temperature.

The present invention provides a process for preparing crystalline mono-HCl form I, comprising:

a) dissolving a compound of formula (XVIII) in an alcoholic solvent containing less than 0.1% w/w water while heating to a solvent of 50 ℃ to 70 ℃, preferably 50 ℃ to 60 ℃;

b) adding hydrochloric acid to the reaction mixture; and

c) the reaction mixture is stirred while maintaining the temperature at 50 ℃ to 70 ℃, preferably 50 ℃ to 60 ℃, more preferably 55 ℃.

In one embodiment, the process for preparing form I as mentioned in the preceding paragraph comprises adding ethanol or methanol at a concentration of 10.3 to 20.6 liters/mole, preferably at a concentration of 10.3 liters/mole.

In another embodiment, the above mentioned process for the preparation of form I comprises dissolving the compound in step a) in a time period of 30 minutes to 3 hours, preferably in a time period of 30 minutes to 45 minutes.

In another embodiment, the above mentioned process for preparing form I comprises adding hydrochloric acid in a concentration of 0.05 to 0.4 equivalents of concentrated HCl, preferably in a concentration of 0.05 to 1.1 equivalents, preferably in a concentration of 0.05 to 0.4 equivalents, in step b).

In another embodiment, the process for the preparation of form I mentioned in paragraphs 1 to 3 above comprises adding hydrochloric acid in a concentration of 0.03 to 0.07 equivalents HCl (1 molar concentration), preferably in a concentration of 0.03 to 0.05 equivalents, in step b).

In another embodiment, the process for the preparation of form I mentioned in paragraphs 1 to 3 above comprises adding hydrochloric acid at a concentration of 0.03 to 0.05 equivalents of HCl in isopropanol in step b).

In another embodiment, the above mentioned process for the preparation of form I comprises dissolving the compound in step c) in a time period of 30 minutes to 3 hours, preferably in a time period of 30 minutes to 45 minutes.

In another embodiment, the above mentioned process for preparing form I comprises stirring the mixture in step c) for 30 minutes to 16 hours, preferably 16 hours.

The present invention further provides a process for preparing a hydrate form comprising:

a) dissolving the compound of formula (XVIII) in a solvent containing more than 5% water, ethanol/water or methanol/water mixture, while heating to 50 ℃ to 70 ℃, preferably 50 ℃ to 60 ℃;

b) adding hydrochloric acid to the reaction mixture; and

c) the reaction mixture is stirred while maintaining the temperature at 50 ℃ to 70 ℃, preferably at 50 ℃ to 60 ℃, more preferably at 55 ℃.

In one embodiment, the process for preparing the hydrate form mentioned in the above paragraph comprises adding ethanol or methanol in a concentration of 10.3 to 20.6 liters/mole, preferably in a concentration of 10.3 liters/mole.

In another embodiment, the above-mentioned method for preparing a hydrate form comprises steps of

a) In (b), the compound is dissolved in a time period of from 30 minutes to 3 hours, preferably in a time period of from 30 minutes to 45 minutes.

In another embodiment, the above-mentioned method for preparing a hydrate form comprises steps of

b) Hydrochloric acid is added in a concentration of 0.05 to 0.4 equivalents of concentrated HCl, preferably in a concentration of 0.05 to 1.1 equivalents, preferably in a concentration of 0.05 to 0.4 equivalents.

In another embodiment, the process for the preparation of a hydrate form as mentioned in paragraphs 1 to 3 above comprises adding hydrochloric acid in a concentration of 0.03 to 0.07 equivalents of HCl (1 molar concentration), preferably in a concentration of 0.03 to 0.05 equivalents, in step b).

In another embodiment, the process for the preparation of a hydrate form as mentioned in paragraphs 1 to 3 above comprises adding hydrochloric acid in a concentration of 0.03 to 0.05 equivalents of HCl in isopropanol in step b).

In another embodiment, the above mentioned process for the preparation of a hydrate form comprises dissolving the compound in step c) in a time period of 30 minutes to 3 hours, preferably in a time period of 30 minutes to 45 minutes.

In another embodiment, the above mentioned process for preparing a hydrate form comprises stirring the mixture in step c) for 30 minutes to 16 hours, preferably 16 hours.

The present invention further provides a pulping process for preparing form I comprising:

-slurrying form II in a solvent selected from ethanol or methanol at a temperature of at least 50 ℃, preferably 70 ℃ or higher; or

-slurrying the mixture of form I and form II in a solvent selected from ethanol or methanol at a temperature of at least 50 ℃, preferably 70 ℃ or higher;

in another embodiment, the above-mentioned pulping process for preparing form I may comprise 10% water, preferably < 2% water, most preferably < 0.07% water.

In another embodiment, the pulping process of making form I further comprises stirring for a period of at least 4 to 7 days.

The present invention further provides a pulping process for preparing a hydrate form comprising:

-slurrying form II in ethanol/water or methanol/water mixture comprising at least 10% water; or

-slurrying the mixture of form I and form II in an ethanol/water or methanol/water mixture comprising at least 10% water;

-slurrying the mixture of form I and form II in an aqueous medium comprising at least 90% water.

In another embodiment, the above-mentioned pulping process for preparing a hydrate form further comprises stirring for a period of at least 4 to 7 days.

In another embodiment, the process for preparing form I further comprises filtering the resulting precipitate after slurrying form II in an alcoholic solvent, or after slurrying a mixture of form I and form II in the solvents referred to above.

In another embodiment, the pulping process for preparing form I further comprises washing the resulting filtered precipitate after the filtration step in the preceding stage, after pulping form II in an alcoholic solvent, or after pulping a mixture of form I and form II in the solvents referred to above, wherein the washing step is carried out with the same solvent used during the pulping step.

To prepare any of the forms of the invention, which are carried out from a solution of the compound of formula (XVIII), one skilled in the art will appreciate that the starting material in solid form has no effect on the final product in solid form, and that control of the resulting solid form is carried out by controlling the process parameters.

The present invention also provides a process for preparing form I as described above, comprising seeding the mixture in form I between steps a) and b).

The present invention also provides the above process for preparing a hydrate form, comprising seeding the mixture in form I between steps a) and b).

The invention also provides a process wherein the crystalline form obtained is isolated by filtration or centrifugation, optionally in combination with washing and drying.

The starting material for the process of the present invention may be any crystalline form of the compound of formula (XVIII).

The term "compounds of the invention" refers to compounds of formula (XI), (XIII), (XVII), (XVIII) or (XIX).

In one embodiment, the solvent used to prepare the crystalline forms of the present invention is a pharmaceutically acceptable solvent. In another embodiment, the solvents used to prepare the crystalline forms of the present invention are pharmaceutically unacceptable solvents because they are found to be useful in the preparation of pharmaceutically acceptable polymorphs.

The process for making the crystalline forms of the invention typically comprises obtaining a crystalline solid material from a solution or dispersion of the compound of formula (XIX) in a solvent medium, or from slurrying of the compound of formula (XIX).

It will be appreciated by those skilled in the art that crystallization-related conditions can be modified to improve the crystallization process or to initiate precipitation without affecting the resulting polymorph form. These conditions may include bringing a solution, dispersion or slurry of a compound of formula (XVIII) or (XIX) and a solvent to the desired concentration, bringing the solution, dispersion or slurry to the desired temperature, adding hydrochloric acid of the desired concentration, adding seed crystals, applying any suitable pressure, removing and/or isolating undesired materials or impurities, drying the crystals formed to give the polymorph in the solid state (if this state is desired).

One preferred way to initiate precipitation is to reduce the solubility of the compounds of the present invention. The solubility of the compound can be reduced, for example, by the addition of an anti-solvent.

Bringing a solution, dispersion or slurry of a compound of the invention and a solvent to the desired concentration does not mean that the concentration of the compound of the invention must be increased. In some cases, it may be preferable to reduce or not alter the concentration of the compounds of the invention. Techniques for obtaining the desired concentration are common in the art, such as evaporation as atmospheric distillation, vacuum distillation, fractional distillation, azeotropic distillation, thin film evaporation, heating, cooling, other techniques known in the art, and combinations thereof. Alternative methods of achieving the desired concentration may also involve saturation of the solution of the compound of the invention and the solvent, for example by adding a sufficient amount of non-solvent to the solution to reach the saturation point. Other suitable techniques for saturating the solution include, for example, introducing additional compounds of the invention into the solution and/or evaporating a portion of the solvent from the solution. As referred to herein, a saturated solution comprises a solution at or above the saturation point (i.e., supersaturated). An almost saturated solution refers to a solution that is near saturation but does not reach the point of saturation.

The method of promoting the crystallization process of the present invention is by seeding with the crystallization of the product or scraping the inner surface of the crystallizer with a glass rod. Alternatively, crystallization may occur automatically without any induction. The present invention includes embodiments in which crystallization of a particular form of a compound of formula (XIX) occurs automatically or is both induced or accelerated (unless such induction or acceleration is critical to obtaining the particular form).

The term "seed crystals" refers to the addition of crystalline material to promote crystallization. The term "seed crystals" refers to the previously obtained powder of the compound of formula (XIX) in crystalline form. Specific seeds or seed materials of the invention that can be used to prepare form I are as follows:

-seeds of a mixture of compound (XIX) of form I and compound of formula (XVIII);

-seeds of form I; or

-seeds of form II.

By bringing the above solution, dispersion or slurry to the desired temperature, the skilled person will be aware of the actions of heating, cooling or placing at ambient temperature. Heating the solution, dispersion or slurry may be necessary to completely dissolve the compounds of the present invention.

Removal and/or isolation of any undesirable materials or impurities may be carried out by purification, filtration, washing, precipitation or similar techniques. The separation can be carried out, for example, by known solid-liquid separation techniques. Filtration procedures known to those skilled in the art can also be used in the methods of the invention. In other methods, filtration can be performed by passing the solution, dispersion or slurry through paper, sintered glass filters or other membrane materials, by centrifugation or using Buchner filters, Rosenmund filters or plate or frame pressure filters. Preferably, in-line filtration or safety filtration can advantageously be put into the above disclosed process to increase the purity of the polymorphic form produced. Furthermore, filtering agents such as silica gel,Diatomaceous earth (dicalite diatemite) and the like may also be used to separate impurities from the associated crystals.

The crystals obtained may also be dried and, if more than one crystallization route is applied, such drying methods may optionally be used for different crystallization routes. The drying procedure includes all techniques known to those skilled in the art, such as heating, application of vacuum, circulating air or gas, addition of a desiccant, freeze drying, spray drying, evaporation, and the like, or any combination thereof.

Methods of crystallizing the polymorph of the compound of formula (XIX) may include various combinations of techniques and variations thereof. Thus and for example, crystallization of a polymorph of a compound of formula (XIX) may be carried out by dissolving, dispersing or slurrying the compound of formula (XIX) in a solvent at a suitable temperature, thereby evaporating a portion of the solvent, increasing the concentration of the compound of formula (XIX) in the above solution, dispersion or slurry, cooling the mixture and optionally washing and/or filtering and drying the resulting compound of formula (XIX) to crystallize. Optionally, the polymorph of the compound of formula (XIX) may be dissolved, dispersed or slurried by placing the compound of formula (XIX) in a solvent medium, cooling the solution, dispersion or slurry and then filtering and drying the resulting polymorph. Another example of preparing a crystalline form of the compound of formula (XIX) may be obtained by placing the compound of formula (XIX) in a solvent medium to become saturated, and optionally filtering, washing and drying the resulting crystals.

The crystallization formation may also comprise more than one crystallization method. In some cases, it may be advantageous to carry out one, two or three additional crystallization steps for different reasons, for example to increase the amount of crystalline form formed.

By dissolving, dispersing or slurrying the compounds of the invention in a solvent, dispersions can be obtained to varying degrees, such as suspensions, slurries or mixtures; or preferably to obtain a homogeneous single phase solution. The term "suspension" refers to a two-phase system consisting of a finely divided solid (amorphous, crystalline form or mixture thereof) dispersed (suspended) in a liquid or suspending medium, usually a solvent. The term "slurry" refers to a suspension formed when a quantity of powder is mixed into a liquid, in which the solids are only slightly dissolved (or not dissolved). "pulping" refers to making a slurry.

Optionally, the solvent medium may contain additives such as one or more dusting agents, surfactants or other additives or mixtures thereof of the type commonly used in the preparation of crystal suspensions and well documented in the literature. The additives can be advantageously used to modify the crystalline shape by increasing the capacity (leniency) and reducing the surface area.

The solid-containing solvent medium may optionally be stirred for a period of time or vigorously agitated using, for example, a high shear mixer or homogenizer, or a combination of these, to produce the desired particle size of the organic compound.

Control of the precipitation temperature and seeding may additionally be used to improve the reproducibility of the crystallization process, the particle size distribution and the form of the product. In this connection, the crystallization may be carried out without seeding with crystals of the compound of formula (XIX), or preferably in the presence of crystals of the compound of formula (XIX), which are introduced into the solution by seeding. Seeding may also be performed several times at various temperatures. The amount of seed material depends on the amount of experiment and can be readily determined by one skilled in the art. Typically, the amount of seed material is about 0.1 to 1 wt% of the amount of crystalline material expected in the reaction.

The crystallization time for each crystallization step will depend on the conditions used, the technique used and/or the solvent used.

After the crystallization transformation, it may be additionally carried out to split the large particles or particle clusters to obtain a desired and uniform particle size. Thus, crystals, powder agglomerates and crude powders of the polymorphic form of the compound of formula (XIX) may optionally be ground and classified by size after undergoing conversion. Milling or grinding refers to physically breaking apart large particles or clusters of particles to reduce the size of the powder particles using methods and devices well known in the art. The resulting particle size may range from millimeters to nanometers, resulting in, i.e., nanocrystals or microcrystals.

The preferred grinding or milling device is a fluid energy mill or a micronizer (microtizer) because of its ability to produce a narrow distribution of small particles. The particle size grinder uses the kinetic energy of collision to divide the particles with a rapidly moving liquid stream between suspended particles. Air-jet is the preferred fluid energy mill. The suspended particles are injected under pressure into the recirculating particle stream. The smaller particles are carried up into the interior of the mill and swept into a vent, such as a cyclone, connected to a particle size classifier. It will be appreciated by those skilled in the art that some of the crystalline forms may be converted to another form during comminution of the particles.

Characterization of the Crystal form

The present invention provides solid mono-HCl salts of compounds of formula (XIX), further characterized in that they are in crystalline form. In one embodiment, the present invention provides a crystalline form of the compound of formula (XIX) selected from form I, form II and a hydrated form. These forms are substantially free of impurities. Suitably, these forms contain no more than 10% impurities, more suitably they contain no more than 5% impurities, still more suitably they contain no more than 1% impurities. The purity of the polymorph can be tested by XPRD and calculated as the area under the peak. These forms are substantially pure. The term "substantially pure" means greater than 90% pure, suitably greater than 95% pure, more suitably greater than 97% pure, and most suitably greater than 99% pure.

The invention further provides a mixture of two or more crystalline forms of the compound of formula (XIX), wherein the crystalline forms are selected from form I, form II and hydrated forms.

In one embodiment, there is provided a mixture comprising form I and form II of the compound of formula (XIX).

In another embodiment, there is provided a mixture comprising form I and a hydrated form of the compound of formula (XIX).

In another embodiment, a mixture is provided comprising a hydrated form of the compound of formula (XIX) and form II.

In another embodiment, there is provided a mixture comprising form I, hydrated form and form II of the compound of formula (XIX).

The invention further provides one or more crystalline forms of the compound of formula (XIX) and an amorphous form of a non-HCl salt of the compound of formula (XIX), wherein the crystalline form is selected from form I, form II, and a hydrated form.

The XPRD intensity peak positions characterizing form I, form II and the hydrated form are expressed in terms of 2-theta angles.

Form I of compound (XIX) is characterized by typical diffraction peaks at the 2-theta position of 15.1 deg. + -0.2 deg., 17.2 deg. + -0.2 deg., 23.4 deg. + -0.2 deg., 24.4 deg. + -0.2 deg., and 27.7 deg. + -0.2 deg.. Form I is further characterized by X-ray powder diffraction peaks at 2-theta positions of 7.6 ° ± 0.2 °, 12.0 ° ± 0.2 ° and 12.5 ° ± 0.2 °.

Form II of compound (XIX) is characterized by typical diffraction peaks at the 2-theta position of 10.8 deg. + -0.2 deg., 13.7 deg. + -0.2 deg., 17.8 deg. + -0.2 deg. and 26.7 deg. + -0.2. Form II is further characterized by X-ray powder diffraction peaks at 7.4 ° ± 0.2 ° and 22.9 ° ± 0.2 ° at the 2-theta position.

The hydrated form of compound (XIX) is characterized by typical diffraction peaks at the 2-theta position of 10.0 ° ± 0.2 °, 13.4 ° ± 0.2 ° and 26.5 ° ± 0.2 °. The hydrated form is further characterized by X-ray powder diffraction peaks at 21.6 ° ± 0.2 ° and 24.9 ° ± 0.2 ° at the 2-theta position.

An X-ray powder diffraction pattern (XPRD) of form I is substantially as shown in figure 2. The X-ray powder diffraction pattern of form II is substantially as shown in figure 7. The X-ray powder diffraction pattern of the hydrated form is substantially as shown in figure 12.

All forms of XPRD data and map representations were obtained using a PhilipsX' PertPRO MPD diffractometer PW3050/60 with generator PW 3040. The instrument was equipped with a Cu LFF X-ray tube PW 3373/00. The compound to be analyzed is coated on a zero background sample loader. The parameters of the instrument are as follows:

-generator voltage: 45kV

Generator amperage: 40mA

-geometry: Bragg-Brentano

-a platform: a rotator platform.

The scan parameters for form I, II and the hydrated form are as follows: ranging from 3 ° to 50 ° 2- θ, continuously scanned at a rate of 0.01675 °/step, 29.845 sec/step. A rotator rotating time of 1 second, a radiation pattern CuK alpha, and a radiation wavelength of

The scan parameters for forms I and II are as follows: ranging from 3 ° to 50 ° 2- θ, continuously scanned at a rate of 0.01675 °/step, 29.845 sec/step. A rotator rotating time of 1 second, a radiation pattern CuK alpha, and a radiation wavelength of

The scan parameters for the hydrated form are as follows: ranging from 3 ° to 50 ° 2- θ, continuously scanned at a rate of 0.01675 °/step, 59.690 sec/step. A rotator rotating time of 1 second, a radiation pattern CuK alpha, and a radiation wavelength of

Incident light path parameters for form I, II and the hydrated form are as follows:

program divergence slit: 15mm

-Soller slit: 0.04rad

-electron beam mask (beam mask): 15mm

-an anti-scatter slit: 1 degree

-an electron beam knife: +

The diffraction optical path parameters for form I, II and the hydrated form are as follows:

-a long anti-scatter-shield: +

-Soller slit: 0.04rad

-a Ni filter: +

-a detector: x' Celerator

Due to experimental differences, such as instrument operation, sample preparation, and the like, the accuracy of the peak positions for the supply form I, II and the hydrated form XPRD was defined to be 0.2 °.

IR absorption peak positions in wavenumbers cm for the characterized form I, II and the hydrated form^-1And (4) showing.

Form I of Compound (XIX) is characterized by an Infrared Spectrum (IR) micro-attenuated reflectance Spectrum at 3119. + -.2 cm^-1、2756±2cm^-1、1634±2cm^-1、1475±2cm^-1、1371±2cm^-1、1333±2cm^-1、1275±2cm^-1、1226±2cm^-1，1128±2cm^-1And 1066cm^-1±2cm^-1With typical absorption bands.

Form II of Compound (XIX) is characterized by an Infrared Spectrum (IR) micro-attenuated reflectance Spectrum at about 3553. + -.2 cm^-1、3203±2cm^-1、3014±2cm^-1And 1541cm^-1±2cm^-1With typical absorption bands.

The hydrate form of Compound (XIX) is characterized by an Infrared Spectrum (IR) micro-attenuated reflectance Spectrum at about 3558 + -2 cm after 42 days of storage at 40 deg.C/75% relative humidity^-1、3238±2cm^-1、1607±2cm^-1And 997cm^-1±2cm^-1With typical absorption bands.

The IR spectrum of form I is substantially as shown in figure 1. The IR spectrum of form II is substantially as shown in figure 6. The IR spectrum of the hydrated form is substantially as shown in figure 11.

IR data and spectral plots were obtained with a Nexus FTIR spectrometer using infrared spectroscopy (IR) micro attenuated total reflectance (microATR). The micro ATR fitting was Harrick split pea with Si crystals. The detector used was a DTGS with KBr window. The form I, II and hydrated form scan parameters were as follows:

-number of scans: 32

-resolution: 2cm^-1

-wavelength range: 4000 to 400cm^-1

-baseline correction: is provided with

-a beam splitter: ge on KBr.

Due to experimental differences, such as instrument handling, sample preparation, and the like, the accuracy of the IR absorption peaks for both the donor form I, II and the hydrate form was defined as 2cm^-1。

Differential Scanning Calorimetry (DSC) endotherm peak positions or ranges characterizing forms I and II are expressed in degrees celsius.

Form I of compound (XIX) is melt decomposed. An exothermic signal was observed at about 216.8 ℃.

Form II of compound (XIX) decomposes by melting at about 197.3 ℃. An exothermic signal was observed at about 203.6 ℃. An additional endothermic signal was observed at about 71.5 ℃ on the DSC curve due to solvent evaporation.

The DSC curve for form I is substantially as shown in figure 3. The DSC curve for form II is substantially as shown in figure 8.

DSC data and graphical representations were obtained using a TA-instruments q1000MTDSC equipped with an RCS cooling unit. The weight of the sample was about 3 mg and transferred to a standard aluminum TA-Instruments sample pan. The sample was scanned at 10 ℃/min from 25 ℃ to a final temperature of 300 ℃. Nitrogen was constantly introduced into the oven at a flow rate of 50 ml/min.

The DSC curve tolerance for forms I and II is defined as 3 ℃ due to experimental differences, such as instrument operation, sample preparation, and the like.

The adsorption-desorption characteristics of form I and form II are expressed in% by mass change.

Form I of compound (XIX) adsorbs up to 0.6% water at high relative humidity, which shows no hygroscopic behavior and remains crystalline during the test.

Form II of compound (XIX) is a hygroscopic product. It adsorbs up to 9.6% water at high relative humidity. This product was completely dry during the desorption cycle and remained crystalline during the experiment.

The ADS/DES curve for form I is substantially as shown in FIG. 5. The ADS/DES curve for form II is substantially as shown in FIG. 10.

The ADS/DES data were obtained using the SMS kinetic vapor sorption model DVS-1 and weight changes were recorded at 25 ℃ atmospheric humidity. The weight of the sample was about 17 mg of form I and 24 mg of form II. The samples were dried under anhydrous nitrogen for 60 minutes. The equilibrium is less than or equal to 0.01%/minute, 15 minutes minimum and 160 minutes maximum. Data intervals were 0.05% or 2.0 minutes.

Relative humidity (%) measurement points were:

a first group: 5. 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 90, 80, 70, 60, 50, 40, 30, 20, 10, 5

Second group: 5. 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 90, 80, 70, 60, 50, 40, 30, 20, 10, 5, 0.

Pharmaceutical use of crystalline forms

The invention further provides form I, form II or a hydrated form of the compound of formula (XIX), a mixture of two or more crystalline forms of the compound of formula (XIX), a mixture of at least form I or a hydrated form of the compound of formula (XIX) and a non-HCl salt amorphous form of the compound of formula (XIX), for use as a pharmaceutical product. In one embodiment, the crystalline form (alone or any mixture thereof) for use as a pharmaceutical product is selected from form I, form II or a hydrated form.

The invention further provides the use of at least form I, form II or a hydrated form of the compound of formula (XIX), a mixture of two or more crystalline forms of the compound of formula (XIX), or a mixture of at least form I or a hydrated form of the compound of formula (XIX) and a non-HCl salt amorphous form of the compound of formula (XIX) for the manufacture of a medicament for the treatment of a condition associated with HDAC. In one embodiment, the crystalline form (alone or in a mixture of any of the above) used in the manufacture of the pharmaceutical product is selected from the group consisting of form I, form II and a hydrated form.

The invention also provides a method of treating a mammal suffering from an HDAC-related condition, comprising administering to a mammal in need thereof at least form I, form II or a hydrated form of the compound of formula (XIX), a mixture of two or more crystalline forms of the compound of formula (XIX), or a mixture of at least form I or a hydrated form of the compound of formula (XIX) and a non-HCl salt amorphous form of the compound of formula (XIX). In one embodiment, the method of treatment comprises administering a crystalline form selected from form I, form II, or a hydrated form (either alone or in any mixture thereof).

As used herein, the terms "histone deacetylase" and "HDAC" refer to any one of a family of enzymes that remove an acetyl group from the epsilon-amino group of a lysine residue at the N-terminus of a histone protein.

Unless the context indicates otherwise, the term "histone" refers to any histone protein, including H1, H2A, H2B, H3, H4, and H5 from any species. Human HDAC proteins or gene products, including but not limited to HDAC-1, HDAC-2, HDAC-3, HDAC-4, HDAC-5, HDAC-6, HDAC-7, HDAC-8, HDAC-9, HDAC-10, and HDAC-11. Histone deacetylases may also be derived from protozoan or fungal sources.

The term "treatment" refers to any treatment of a pathological condition in a mammal (particularly a human being) and includes one or more of the following actions:

(i) preventing the onset of a pathological condition in a subject predisposed to the condition but not yet diagnosed with the condition, and thus, treatment includes prophylactic treatment of the condition;

(ii) inhibiting the pathological symptoms, i.e. arresting their development;

(iii) relieving pathological symptoms, namely causing the recovery of pathological symptoms; or

(iv) Alleviating the symptoms mediated by the pathological symptoms.

The term "polymorph(s) of the invention" refers to at least form I, form II or hydrated form of the compound of formula (XIX), a mixture of two or more crystalline forms of the compound of formula (XIX), or a mixture of at least form I or hydrated form of the compound of formula (XIX) and a non-HCl salt amorphous form of the compound of formula (XIX).

The present invention provides methods for inhibiting the abnormal growth of cells, including transformed cells, by administering an effective amount of a polymorph of the invention. Abnormal growth of a cell means that cell growth is not dependent on normal regulatory mechanisms (e.g., loss of contact inhibition). This includes inhibiting tumor growth both directly by causing growth arrest, terminal differentiation and/or apoptosis of cancer cells, and indirectly by inhibiting tumor angiogenesis.

The present invention also provides methods of inhibiting tumor growth by administering an effective amount of a polymorph of the present invention to a subject in need thereof, such as a mammal (more particularly a human). In particular, the present invention provides methods for inhibiting tumor growth by administering an effective amount of the polymorphic forms of the present invention. Examples of tumors that can be inhibited include, but are not limited to, lung cancer (e.g., adenocarcinoma and including non-small cell lung cancer), pancreatic cancer (e.g., pancreatic cancer such as exocrine pancreatic cancer), colon cancer (e.g., colorectal adenocarcinoma such as colon adenocarcinoma and colon adenoma), prostate cancer including advanced disease, hematopoietic tumors of the lymphatic system (e.g., acute lymphocytic leukemia, B-cell lymphoma, Burkitt's lymphoma), Hodgkin's disease and non-Hodgkin's disease, myelogenous leukemia (e.g., Acute Myelogenous Leukemia (AML)), thyroid follicular cancer, myelodysplastic syndrome (MDS), tumors of mesenchymal origin (e.g., fibrosarcoma, rhabdomyosarcoma), melanoma, teratocarcinoma, neuroblastoma, glioma, benign tumors of the skin (e.g., keratoacanthoma), breast cancer (e.g., advanced breast cancer), renal cancer, ovarian cancer, Bladder cancer and epithelial cancer.

The present invention further provides a pharmaceutical composition comprising at least form I, form II or a hydrated form of the compound of formula (XIX), a mixture of two or more crystalline forms of the compound of formula (XIX), or a mixture of at least form I or a hydrated form of the compound of formula (XIX) and a non-HCl salt amorphous form of the compound of formula (XIX), and a pharmaceutically acceptable excipient. In one embodiment, the pharmaceutical composition comprises a crystalline form selected from form I, form II and a hydrated form (either alone or in a mixture of any of the foregoing).

The pharmaceutical compositions may be prepared as medicaments for oral, parenteral (including subcutaneous, intramuscular and intravenous), rectal, transdermal, buccal or intranasal administration. Forms suitable for oral administration include powders, granules, pellets, tablets, compressed or coated pills, dragees, sachets, hard or gelatin capsules, syrups and suspensions. Suitable parenteral administration forms include aqueous or non-aqueous solutions or emulsions, while suitable administration forms for rectal administration include suppositories with hydrophilic or hydrophobic carriers. For topical administration, the present invention provides suitable transdermal delivery systems known in the art, and for intranasal delivery, suitable aerosol delivery systems known in the art. The most preferred route of the invention is oral, although the most suitable administration in any given case will depend on the nature and severity of the symptoms to be treated.

The dosages may be conveniently presented in unit dosage form and prepared by any of the methods well-known in the art of pharmacy. In addition, the dosage form may be presented over the course of a day in one, two, three or four or more sub-doses, for administration at suitable intervals. The unit dosage used is preferably from about 1 mg to about 1000 mg of the base equivalent of the compound of formula (XIX), more preferably from about 5 to about 400 mg.

The pharmaceutical compositions of the invention include form I, form II or a hydrated form of the compound of formula (XIX). The pharmaceutical composition may comprise only a single form of form I, form II or hydrated form of the compound of formula (XIX), or a mixture of forms of the compound of formula (XIX), with or without the non-HCl salt amorphous form of the compound of formula (XIX). The pharmaceutical composition may comprise one or more excipients or adjuvants in addition to one or more active ingredients. The choice of excipients and the amounts used can be readily determined by the pharmaceutical formulator according to experience and consideration of standard manufacturing procedures and reference procedures in the art.

Examples of suitable excipients are acacia, magnesium oxide, magnesium carbonate, potassium phosphate, lactose, glucose or starch, in particular corn starch. Suitable oily vehicles or solvents are vegetable or animal oils, for example sunflower oil or cod liver oil. Suitable solvents for the aqueous or alcoholic solution are water, ethanol, sugar solutions or mixtures thereof. Polyethylene glycol and polypropylene glycol may also be used as additional adjuvants for other forms of administration.

For subcutaneous or intravenous administration, the polymorph of the invention is added, if desired, to a solution, suspension or emulsion together with the usual substances, such as solubilizers, emulsifiers or other auxiliaries. The polymorph of the invention can also be lyophilized and the lyophilizate obtained used, for example, for the manufacture of injection or infusion preparations. Suitable solvents are, for example, water, physiological saline solution or alcohols such as ethanol, propanol, glycerol, in addition sugar solutions such as glucose or mannitol solutions, or also mixtures of the various solvents mentioned.

Pharmaceutical compositions suitable for administration in aerosol or spray form are, for example, solutions, suspensions or emulsions of the polymorph of the invention in a pharmaceutically acceptable solvent such as ethanol or water or a mixture of such solvents. The formulations may, if desired, additionally contain other pharmaceutical auxiliaries, such as surfactants, emulsifiers and stabilizers, and also propellants. Such preparations usually contain the active compound in a concentration of about 0.1 to 50%, in particular about 0.3 to 3% by weight.

It will be appreciated that in addition to the ingredients particularly mentioned above, the pharmaceutical compositions of the present invention may include other agents conventional in the art having regard to the type of formulation in question, for example pharmaceutical compositions suitable for oral administration may include flavouring or taste masking agents.

As a further aspect of the invention, combinations of form I, form II or a hydrated form of the compound of formula (XIX), a mixture of two or more crystalline forms of the compound of formula (XIX), or a mixture of at least form I or a hydrated form of the compound of formula (XIX) and a non-HCl salt amorphous form of the compound of formula (XIX) with a further anti-cancer agent, in particular for use as a medicament, more in particular for the treatment of cancer or a related disease, are also envisaged.

For the treatment of the above symptoms, the polymorph of the invention can advantageously be combined with one or more pharmaceutical agents, more particularly with other anticancer agents. Examples of anti-cancer agents include, but are not limited to:

-platinum coordination compounds, such as cisplatin (cissplatin), carboplatin (carboplatin) or oxaliplatin (oxalyplatin);

-taxoids (taxanes), such as paclitaxel (paclitaxel) or docetaxel (docetaxel);

topoisomerases i (topoisomerases i) inhibitors, for example camptothecin (camptothecin) compounds such as irinotecan (irinotecan) or topotecan (topotecan);

inhibitors of topoisomerase II such as anti-tumour epipodophyllotoxins (epipodophyllotoxins) or podophyllotoxin (podophyllotoxin) derivatives, such as etoposide (etoposide) or teniposide (teniposide);

anti-tumour vinca alkaloids (vinca alkaloids) such as vinblastine (vinblastine), vincristine (vinchristine) or vinorelbine (vinorelbine);

antineoplastic nucleoside derivatives such as 5-fluorouracil (5-fluorouracil), folinin (leucovorin), gemcitabine (gemcitabine) or capecitabine (capecitabine);

alkylating agents such as nitrogen mustards or nitrosoureas, for example cyclophosphamide (cyclophosphamide), chlorambucil (chlorambucil), carmustine (carmustine), thiotepa (thiotepa), mevastatin (phalan) or lomustine (lomustine);

antineoplastic anthracycline derivatives, such as daunorubicin (daunorubicin), doxorubicin (doxorubicin), doxil, idarubicin (idarubicin) or mitoxantrone (mitoxantrone);

molecules targeting IGF-1 receptors, such as picropodophilin (picropodophilin);

-a tetracacin (tetracaine) derivative such as tetracacin a (tetracarcin a);

glucocorticoids such as prednisone (prednisone);

antibodies such as trastuzumab (trastuzumab) (HER2 antibody), rituximab (rituximab) (CD20 antibody), gemtuzumab (gemtuzamab), cetuximab (cetuximab), pertuzumab (pertuzumab), or bevacizumab (bevacizumab);

-estrogen receptor antagonists or selective estrogen receptor modulators, such as tamoxifen (tamoxifen), fulvestrant (fulvestrant), toremifene (toremifene), droloxifene (droloxifene), valdecoxix (faslodex) or raloxifene (raloxifene);

aromatase inhibitors such as exemestane (exemestane), anastrozole (anastrozole), letrozole ((letrozole) and vorozole (vorozole);

differentiating agents such as retinoids, vitamin D or retinoic acid and Retinoic Acid Metabolic Blockers (RAMBA), such as isotretinoin (accutane);

DNA methyltransferase inhibitors, such as azacytidine (azacytidine) or decitabine (decitabine);

antifolates such as pemetrexed disodium (premetrexed disodium);

antibiotics such as actinomycin D (antinomycin D), bleomycin (bleomycin), mithramycin C (mitomycin C), dactinomycin (dactinomycin), carminomycin (carminomycin) or daunomycin (daunomycin);

antimetabolites such as clofarabine (chlorofarabine), aminopterin (aminopterin), cytarabine (cytosine arabinoside) or methotrexate (methotrexate);

apoptosis inducers and antiangiogenic agents, for example Bcl-2 inhibitors such as YC 137, BH 312, ABT 737, gossypol (gossypol), HA 14-1, TW 37 or decanoic acid;

tubulin binding agents, such as combretastatin (combestin), colchicines (colchicines) or nocodazole (nocodazole);

kinase inhibitors, such as, for example, flopiridol (flavoperidol), imatinib mesylate (imatinib), erlotinib (erlotinib) or gefitinib (gefitinib);

farnesyl transferase (farnesyltransferase) inhibitors such as tipifarnib;

-Histone Deacetylase (HDAC) inhibitors, such as sodium butyrate, suberoylanilide hydroxyacyl acid (SAHA), depsipeptide (depsipeptide) (FR 901228), NVP-LAQ824, R306465, JNJ-26481585 or trichostatin a (trichostatin a);

-inhibitors of the ubiquitin-proteasome pathway, such as PS-341, mln.41 or bortezomib (bortezomib);

-Yondelis；

telomerase inhibitors, such as tylostatin (telomestatin);

matrix metalloproteinase inhibitors, such as batimastat (batimastat), marimastat (marimastat), prinostat (prinostat) or mestat (metastat).

The components of the combination of the invention, i.e. the other pharmaceutical agents and the polymorph of the invention, may be formulated in a variety of pharmaceutical forms for administration purposes, as regards their useful pharmacological properties. Such components may be formulated separately into individual pharmaceutical compositions or as a single pharmaceutical composition containing both components.

One embodiment of the present invention thus also relates to pharmaceutical compositions comprising other agents and polymorphs of the present invention, together with one or more pharmaceutical carriers.

The invention further relates to the use of a combination of the invention for the manufacture of a pharmaceutical composition for inhibiting the growth of tumor cells.

The invention further relates to products containing the polymorph of the invention as a first active ingredient and an anti-cancer agent as a second ingredient, as a combined preparation for simultaneous, separate or sequential use in the treatment of a patient suffering from cancer.

The other agents and polymorphs of the present invention may be administered simultaneously (e.g. in separate or single compositions) or sequentially in either order. In the latter case, both compounds are administered over a period of time in an amount and manner sufficient to ensure that a beneficial or synergistic effect is achieved. It will be appreciated that the preferred method and order of administration and the individual dosages and schedules of the components of the combination will depend on the particular other agent and polymorph desired to be administered, its route of administration, the particular tumor desired to be treated and the particular host desired to be treated. The optimal method and order of administration, as well as the dosage and regimen, can be readily determined by one of skill in the art using available methods and in light of the data presented herein.

The platinum coordination compound is advantageously present in an amount of 1 to 500 milligrams per square meter of body surface area (mg/m)²) Is administered, e.g. 50 to 400mg/m²Particularly about 75mg/m for cisplatin per treatment course²In a dose of about 300mg/m with carboplatin²。

The taxoid compound is advantageously present in an amount of 50 to 400 milligrams per square meter of body surface area (mg/m)²) Is administered, e.g. 75 to 250mg/m²In particular, paclitaxel is administered at a dose of about 175 to 250mg/m per treatment course²And about 75 to 150mg/m of docetaxel²。

The camptothecin compound is advantageously present in an amount of 0.1 to 400 milligrams per square meter of body surface area (mg/m)²) Is administered, e.g., in a dose of 1 to 300mg/m²In particular, about 100 to 350mg/m per treatment course of irinotecan²And about 1 to 2mg/m for topotecan²。

The antitumor podophyllotoxin derivative is advantageously present in an amount of 30 to 300 milligrams per square meter of body surface area (mg/m)²) Is administered, e.g. 50 to 250mg/m²In particular to etoposide per treatment courseAbout 35 to 100mg/m²And about 50 to 250mg/m of p-teniposide²。

The antitumor alkaloid is advantageously present in an amount of 2 to 30 milligrams per square meter of body surface area (mg/m)²) In particular to about 3 to 12mg/m of vinblastine per treatment course²About 1 to 2mg/m of vincristine²And about 10 to 30mg/m of vinorelbine²The dosage of (a).

The antitumor nucleoside derivative is advantageously present in an amount of 200 to 2500 milligrams per square meter of body surface area (mg/m)²) Is administered, e.g. 700 to 1500mg/m²In particular 200 to 500mg/m per course of treatment for 5-FU²The dose of (a) to gemcitabine, about 800 to 1200mg/m²In a dosage of about 1000 to 2500mg/m for capecitabine²。

Alkylating agents such as nitrogen mustards or nitrosoureas are advantageously used in amounts of 100 to 500 milligrams per square meter of body surface area (mg/m)²) Is administered, e.g. 120 to 200mg/m²In particular, about 100 to 500mg/m per treatment course of cyclophosphamide²In a dosage of about 0.1 to 0.2mg/kg for chlorambucil and about 150 to 200mg/m for carmustine²At a dosage of about 100 to 150mg/m for lomustine²The dosage of (a).

The antitumor anthracycline derivative is advantageously present in an amount of 10 to 75 milligrams per square meter of body surface area (mg/m)²) Is administered, e.g., 15 to 60mg/m²In particular, about 40 to 75mg/m of doxorubicin per course of treatment²In a dose of about 25 to 45mg/m for daunorubicin²And about 10 to 15mg/m of idarubicin²The dosage of (a).

The antiestrogen agent is advantageously administered in a dosage of about 1 to 100mg per day, depending on the particular agent and the condition being treated. Tamoxifen is advantageously administered orally at a dose of 5 to 50mg, preferably 10 to 20 mg twice daily, for a sufficient time for treatment to reach and maintain therapeutic efficacy. Toremifene (toremifene) is advantageously administered orally at a dose of about 60mg once a day for a sufficient time to achieve and maintain therapeutic efficacy. Anastrozole is advantageously administered once daily in a dose of about 1 mg. Droloxifene is advantageously administered orally once daily in a dose of about 20-100 mg. Raloxifene is advantageously administered orally once daily in a dose of about 60 mg. Exemestane is advantageously administered orally once daily in a dose of about 25 mg.

The antibody is advantageously present in an amount of about 1 to 5 milligrams per square meter of body surface area (mg/m)²) Or if different, as known in the art. Trastuzumab is advantageously present in an amount of 1 to 5 milligrams per square meter of body surface area (mg/m)²) Is administered, in particular, 2 to 4mg/m per course of treatment²。

These doses may be administered, for example, once, twice or more per course of treatment, which may be repeated, for example, every 7, 14, 21 or 28 days.

The polymorph of the invention may conveniently be stored in packaging materials which protect it from mechanical, environmental, biological or chemical damage or degradation. Conditioning the drug substance can be achieved by using a moisture impermeable packaging material, such as a closed airtight bag. Conditioning pharmaceutical products, such as tablets, capsules, can be achieved using, for example, aluminum blisters.

Experimental part

The following examples are intended to illustrate the invention and are not intended to limit the scope of the invention.

Example 1: preparation of intermediate (I)

a) 4-Piperidinemethylamine (2.6 mol) and ethyl acetate (5.2 l) were placed in an inert reactor (20 l) and warmed to 45 ℃. 4-nitrobenzaldehyde (2.7 moles) was added and the reaction mixture was stirred at 45 ℃ for 2 hours. The reaction was cooled to 0 ℃ and diisopropylethylamine (6.6 moles) was added to give solution 1.

b) Ethyl 2- (methylthio) -5-pyrimidinecarboxylate (2.7 mol) and ethyl acetate (2.6 l) were placed in an inert reactor and cooled to 0 ℃. A solution of m-chloroperoxybenzoic acid (1.2 moles) in ethyl acetate (2.6 liters) was added over a period of 1 hour at a temperature of 0 ℃ to 5 ℃. The reaction mixture was stirred at 0 ℃ for 30 minutes to give solution 2.

c) Solution 2 was added to solution 1 over a period of 1 hour at a temperature of 0 ℃ to 5 ℃. The reaction mixture was left at room temperature overnight. The mixture was acidified to pH2 with 640 ml of concentrated hydrochloric acid in 10 l of water. The aqueous layer was collected and washed with 1 liter of ethyl acetate. The water layer was collected and 1 liter of dichloromethane was added. The mixture was basified with 450 ml 50% sodium hydroxide to pH 10. The reaction mixture was stirred at room temperature for 30 minutes. The organic layer was collected to give fraction 1. The aqueous layer was further extracted with 2 liters of dichloromethane and the organic layer was collected to give fraction 2. Fractions 1 and 2 were combined and dichloromethane was evaporated, yielding 511.25 g (1.93 mol) of intermediate (I) (74% yield).

Example 2: preparation of intermediate (XIII)

a) Intermediate (I) (0.97 mol) and toluene (4.5 l) were charged to an inert reactor (20 l). 1-methyl-1H-indole-3-carbaldehyde (0.97 mol) was added to the reaction mixture at room temperature. The reaction mixture was warmed to reflux temperature, refluxed overnight and cooled to room temperature. Methanol-degenerate ethanol (1.5 l) was added to give solution 3.

b) Sodium tetrahydroborate (55.2 g) and toluene (1.5 l) were charged to an inert reactor (20 l). The mixture was brought to 10 ℃ with constant stirring. Solution 3 was added to the mixture over a period of 1 hour at a temperature of #10 ℃. The mixture was stirred at a temperature of #10 ℃ for 1 hour. The reaction mixture was allowed to reach room temperature. Acetone (8.79 mol) was added over 30 minutes. The reaction mixture was stirred for 4 hours. Water (5.1 l) was added dropwise to the reaction mixture over a period of 15 minutes. The reaction mixture was stirred at room temperature for 1 hour. The aqueous layer was discarded and the organic layer was washed twice with 300 g of sodium bicarbonate in 4.1 l of water. The organic layer was filtered over magnesium sulfate and evaporated to give fraction 3 (after evaporation, 397 g of residue).

c) Ethanol (5 l) degenerated with 2% methyl ethyl ketone was added to fraction 3 at room temperature. Concentrated acetone (5 l) and 0.5 l water were added at room temperature and the mixture was subsequently warmed to 50 ℃. A mixture of fumaric acid (0.97 mol), ethanol (1.4 l) degenerated with 2% methyl ethyl ketone, acetone (1.4 l) and 140 ml of water was prepared to give solution 4. Solution 4 was added to the reaction mixture over a period of 2 hours at a temperature of 50 ℃. The reaction mixture was stirred at 50 ℃ for 2 hours, cooled 4 hours to room temperature and stirred at room temperature overnight. The precipitate was collected and subsequently washed with 1.4 l ethanol degenerated with 2% methyl ethyl ketone, 1.4 l concentrated acetone and 140 ml water. The precipitate was dried at 50 ℃ overnight to give 371 g (0.7 mol) of intermediate (XIII) (73% yield).

Example 3: preparation of intermediate (XVII)

A four-necked flask (2L) was charged with intermediate (XIII) (100 g; 191.0 mmol). Water (3L/mol-pure limiting reagent; 573.0 ml) and 2-methyltetrahydrofuran (2.2L/mol-pure limiting reagent; 420.2 ml) were added. After stirring, 50% sodium hydroxide (2.5 mol/mol-pure-limiting reagent; 477.5 mmol; 25.13 ml) was added. The reaction mixture was stirred at room temperature for another 40-60 minutes, after which the reaction was allowed to settle (settle). The upper organic layer was collected and washed with water (2 l/mol-pure-limiting reagent; 382.0 ml). Water (1.5L/mol-pure-limiting reagent; 286.5 ml) and sodium hydroxide (3 mol/mol-pure-limiting reagent; 573.0 mmol; 45.84 g) were added to the organic layer. The reaction mixture was warmed to 80 ℃ and stirred for 16 hours. The reaction mixture was cooled to room temperature and the lower aqueous layer was collected. Isopropanol (90 ml; 1.177 mol) was added and the mixture was cooled to 10 ℃ in an ice bath. The reaction mixture was acidified to pH1(pH 13.8: dark green solution; pH7.5 dark pink; pH 4.7: pink solution, white precipitate) with concentrated hydrochloric acid (5M/M-pure-restriction reagent; 954.9M; 100.9 g). The reaction mixture was stirred at 10 ℃ for 4 hours. The white precipitate was filtered, washed 4 times with water and dried under vacuum at 40 ℃ to give 84 g of intermediate (XVII) (yield: 97%).

Example 4: preparation of intermediate (XVIII)

a) A four-necked flask (1L) was charged with 0.093 mol of intermediate (XVII) and 220 mL of ethyl acetate was added. The reaction mixture was stirred and 5 ml of water were added to obtain solution 5. A250 ml flask was charged with a solution of 0.122 mole of N- (ethylcarbonylimino) -N, N-dimethyl-1, 3-propanediamine, monohydrochloride (EDC) in 130 ml of ethanol and the reaction mixture was stirred to give solution 6. O- (tetrahydro-2H-pyran-2-yl) -hydroxylamine (0.123 mol) was added to solution 5 and the addition funnel was washed with 26 ml of ethyl acetate. Immediately thereafter, 200 ml of solution 6 were added to the reaction mixture containing solution 5 over a period of 1 hour and 30 minutes (when 90% of solution 6 was added, the reaction mixture became homogeneous and the desired product crystallized out). The reaction mixture was stirred at room temperature for 5 hours. The precipitate was filtered off and washed with 55 ml of ethyl acetate and dried under vacuum at 50 ℃ for 16 hours to give 35.7 g (0.07 mol) of intermediate (XVIII) (yield: 71%).

b) A four-necked flask (1 liter) was charged with 0.073 mol of intermediate (XVIII) under a nitrogen atmosphere. N, N dimethylacetamide (377 ml) and 377 ml methyl isobutyl ketone were added and the mixture was warmed to 70 ℃. The reaction mixture was stirred at 70 ℃ for 5 hours, then cooled to 25 ℃ over a period of 1 hour, and then stirred at 25 ℃ for an additional hour. The precipitate was filtered and then washed with 94 ml of a mixture of N, N dimethylacetamide and methyl isobutyl ketone, then with 150 ml of a slurry of methyl isobutyl ketone, then with 150 ml of a displacement wash of methyl isobutyl ketone. The precipitate was dried under vacuum at 50 ℃ for 2 days to give 33.4 g of purified intermediate (XVIII) (yield: 89%).

Example 5: preparation of JNJ-26481585HCl salt crystalline form I

a) An inert four-necked flask (0.5 liter) was charged with 0.03 mole of purified intermediate (XVIII). Ethanol (300 ml) was added (typical water content 0.07% (w/w)). The reaction mixture was stirred and warmed to 57-60 ℃. Intermediate (XVIII) JNJ-26481585.HCl (30 mg) form I was seeded with 30 mg. Concentrated hydrochloric acid (0.05 mol%) was added to the reaction mixture at 57 ℃ and the reaction mixture was stirred for 16 hours. The precipitate was filtered at 50 ℃ and washed 3 times with 20 ml of ethanol to give 10 g of JNJ-26481585HCl salt in crystalline form I.

b) An inert four-necked flask (50 ml) was charged with 2.6 g of the crystalline form I of JNJ-26481585HCl salt obtained in step a). Ethanol (20 ml) was added. The reaction mixture was stirred under nitrogen and in the dark and warmed to 50 ℃. The reaction mixture was stirred at 50 ℃ for 12 hours, cooled to 40 ℃ over a period of 1 hour and filtered. The precipitate was washed once with 20 ml ethanol and twice with 20 ml acetone. The product was then dried under vacuum at 50 ℃ for 16 h, yielding 2 g (80%) of purified JNJ-26481585HCl salt crystalline form I.

Example 6: transformation of mixtures of polymorphs I and II using a slurry procedure

a) Preparation of the slurry

About 25 mg of form I and about 25 mg of form II were weighed into a vial. About 0.2 ml ethanol was added and the vial was closed. Three vials were prepared and each vial was stored at different temperatures, 4 ℃ (refrigerator), 40 ℃ and 70 ℃ for 4 days.

The process was repeated for a slurry of ethanol/water (90/10, vol/vol%) and water. The slurries were stored at different temperatures for 4 days and 7 days. After storage, the vial was opened and the sample dried by spreading several milligrams of the slurry on filter paper.

b) Analytical technique (powder XRD)

All the obtained fractions were analyzed using powder XRD.

X-ray powder diffraction (XRPD) analysis was carried out in a Philips X' PertPRO MPD diffractometer PW3050/60 with generator PW 3040. The instrument was equipped with a Cu LFF X-ray tube PW 3373/00. The compound was spread on a zero background sample carrier.

Parameters of the instrument

Generator voltage: 45kV

Generator amperage: 40mA

Geometry: Bragg-Brentano

Platform: a rotator platform.

Measurement conditions

Scanning mode: continuous

Scanning range: 3 to 50 DEG 2 theta

Step size: 0.01675 degree/step

Counting time: 29.85 seconds per step

Rotator rotation time: 1 second

Type of radiation: CuKa

Wavelength of radiation:

incident light path diffraction light path

Program divergence slit: 15mm long anti-scatter-cover: +

Soller slit: 0.04rad Soller slit: 0.04rad

Electron beam mask: 15mm Ni Filter: +

Anti-scattering slit: 1 ° detector: x' Celerator

An electron beam knife: +

c) Results

The results obtained in the slurry conversion study after storage in ethanol for 4 days and 7 days are collected in table a below.

Slurry time and temperature	Ethanol after 4 days	Ethanol after 7 days
			Refrigerator at 4 deg.C 40 deg.C 70 deg.C	Solvate + mixture of form I + form II: (*) Form I	Solvate + mixture of form I form II

The XRD pattern of the solvated form is comparable to that of the hydrate.

The results obtained in the slurry conversion study after 4 days and 7 days of storage in ethanol/water (90/10, v/v%) are collected in table B below.

Slurry time and temperature	Ethanol/water (90/10, vol/vol%)	Ethanol/water (90/10, vol/vol%)
			Refrigerator at 4 deg.C 40 deg.C 70 deg.C	Mixture of hydrate + form I + mixture of form II ((II))*) Mixtures of hydrate + form I	Mixture of hydrate + form IMixture form I + micro hydrate

(^*) The solvent in the slurry was completely evaporated. After four days of storage, 0.2 ml of solvent was added to the mixture.

The results obtained in the slurry conversion study after storage in water for 4 days are collected in table C below.

Slurry time and temperature	Water after 4 days
		40 ℃ in a 4 ℃ refrigerator	Hydrate

70℃	Hydrate of calcium and magnesium

These hydrated samples (e.g., aqueous slurries) were stored with 0.1 ml of ethanol at various temperatures, 40 ℃, 50 ℃ and 70 ℃ for 3 days.

The hydrated samples remained hydrated after 3 days of storage at 40 ℃ and 50 ℃.

The hydrated samples completely liquefied (oil) after 3 days of storage at 70 ℃.

Example 7: stability of form I

a) Compound information

The chemical formula is shown as the figure:

hydrochloride (1: 1)

Chemical name: n-hydroxy-2- [4- [ [ [ (1-methyl-1H-indol-3-yl) methyl ] amino ] methyl ] -1-piperidinyl ] -5-pyrimidinecarboxamide hydrochloride

The molecular formula is as follows: c₂₁H₂₆N₆O₂.HCl

Molecular weight: 430.94

b) Adsorption/desorption study

Adsorption and desorption of water at 25 ℃ under different relative humidities was analyzed by 17 mg of form I.

The change in weight as a function of relative humidity was recorded.

The results are shown in FIG. 4.

The form I fraction absorbs at most 0.6% water at high relative humidity, which shows no hygroscopic behaviour and remains crystalline during the test.

c) Solubility in water

The water solubility of form I was measured in solvents of different pH. Excess solute was equilibrated with solvent at 20 ℃ for 24 hours. After removal of the insoluble compounds, the concentration of the solution was determined using UV spectroscopy.

The solubilities are listed in table D below:

solvent(s)	Solubility (mg/ml solution)
		Form I
Water 0.01N HCl0.001N HCl buffer pH2 (citric acid/NaOH/HCl) buffer pH4 (citric acid/HCl/NaOH) buffer pH6 (citric acid/NaOH) buffer pH8 (boric acid/HCl/NaOH) buffer pH10 (boric acid/KCl/NaOH)	1.4(pH4.5)1.4(pH2.0)1.5(pH2.9)0.95(pH2.0)1.2(pH3.9)1.5(pH6.0)1.3(pH7.8)1.3(pH9.8)

d) Stability of crystallography

The stability of the crystal structure of form I was investigated after storing the compounds at Room Temperature (RT), at < 5%, 56% and 75% Relative Humidity (RH), 50 ℃ and 40 ℃/75% RH for six weeks under open conditions.

Samples were analyzed by Thermogravimetry (TGA), Differential Scanning Calorimetry (DSC), X-ray powder diffraction (XRPD), and infrared spectroscopy (IR).

The test results are shown in table E below.

-Ref: same as the reference

Cryst.: crystal

Form I melts down and therefore no heat of fusion is reported.

Form I is crystallographically stable.

No change was observed after storage under different conditions.

The IR spectra, XRD patterns and DSC curves remained the same before and after storage.

e) Chemical stability

Form I was stored under different open conditions for 1, 4 and 8 weeks. These conditions were 40 ℃/75% RH, 50 ℃, RT/< 5% RH, RT/56% RH, RT/75% RH and 0.3da ICH light.

After storage the compounds were analyzed by HPLC and visual inspection.

The test results are shown in table F below.

Form I showed sensitivity to light due to the sum of the increase in impurities after storage under 0.3da ICH light conditions.

Example 8: stability of form II

a) Compound information

The chemical formula is shown as the figure:

hydrochloride (1: 1)

Chemical name: n-hydroxy-2- [4- [ [ [ (1-methyl-1H-indol-3-yl) methyl ] amino ] methyl ] -1-piperidinyl ] -5-pyrimidinecarboxamide hydrochloride

The molecular formula is as follows: c₂₁H₂₆N₆O₂.HCl

Molecular weight: 430.94

b) Adsorption/desorption study

Adsorption and desorption of water at 25 ℃ under different relative humidities was analyzed by about 24 mg of form II.

The change in weight as a function of relative humidity was recorded.

The results are shown in FIG. 9.

Form II weight loss was recorded as 1.67% during the initial drying step. The resulting dried product is hygroscopic. It absorbs up to 9.6% water at high relative humidity.

During the desorption cycle, the product was completely dry and remained crystalline during the experiment.

c) Stability of crystallography

The stability of the form II crystal structure was studied after storing the compounds at Room Temperature (RT) under < 5%, 56% and 75% Relative Humidity (RH), 50 ℃ and 40 ℃/75% RH for six weeks under open conditions.

Samples were analyzed by Thermogravimetry (TGA), Differential Scanning Calorimetry (DSC), X-ray powder diffraction (XRPD), and infrared spectroscopy (IR).

The test results are shown in table G below.

-Ref: same as the reference

Cryst.: crystal

Form II is melt decomposed and therefore no heat of fusion is reported.

The additional endothermic signal in the DSC curve is due to solvent evaporation.

Form II is crystallographically unstable.

After storage under different humidity conditions, changes were observed.

The starting material differed from the IR spectrum and XRD pattern after storage at RT/56% RH, RT/75% RH and 40 ℃/75% RH.

The change after storage at RT/56% RH, RT/75% RH and 40 ℃/75% RH is due to water absorption.

d) Chemical stability

Form II was stored under different open conditions for 1, 4 and 8 weeks. These conditions were 40 ℃/75% RH, 50 ℃, RT/< 5% RH, RT/56% RH, RT/75% RH and 0.3da ICH light.

After storage the compounds were analyzed by HPLC and visual inspection.

The results of the tests are shown in table H below.

The chemical stability study of R425754 resulted from the following observations:

r425754 was shown to be sensitive to light due to the sum of the increase in impurities after storage under 0.3da ICH light conditions.

A color change from white to or orange-brown was also observed after 0.3da ICH light storage and from white to pink after storage under RT/56% RH, RT/75% RH and 40 ℃/75% RH humidity conditions and at elevated temperatures of 50 ℃.

Claims (4)

1. A compound of the formula (XI),

or an addition salt thereof.

2. A process for preparing a compound of claim 1 comprising reacting an intermediate of formula (IX) with an intermediate of formula (X) in the presence of a suitable solvent

3. Use of a compound of claim 1 in a process for the preparation of a compound of formula (XVIII), said process comprising

a) Reacting an intermediate of formula (VIII) with an intermediate of formula (XI) in the presence of a suitable solvent,

b) reacting the intermediate of formula (I) with the intermediate of formula (II) in a suitable solvent, followed by reduction and salt formation to give an intermediate of formula (XIII),

c) converting the intermediate of formula (XIII) to an intermediate of formula (XVII) by base neutralization, basic hydrolysis and acidification with hydrochloric acid, and

d) reacting an intermediate of formula (XVII) with O- (tetrahydro-2H-pyran-2-yl) -hydroxylamine in the presence of a suitable coupling agent

4. Use according to claim 3, wherein in the intermediate (XVII) of step d) the amount of water is between 15 and 25% v/v.