WO2017125097A1 - Crystalline forms of (3r)-3-cyclopentyl-3-[4-(7h-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile salts and preparation thereof - Google Patents

Abstract

The present invention relates to crystalline forms of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-1-yl]propanenitrile salts of Formula I, wherein HX represents at least one acid component, preferably hydrochloric, phosphoric acid, fumaric acid and L-tartaric acid. The invention also relates to the processes for the preparation thereof as well as said use thereof in phamaceutically acceptable compositions. Use of said crystalline forms of ruxolitinib and manufactured salts in the preparation of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-1-yl]propanenitrile in the free form or in the form of any phamaceutically acceptable salt thereof is also part of this invention.

Description

Crystalline forms of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile salts and preparation thereof

Field of the Invention The present invention relates to crystalline forms of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile salts of Formula I,

H-X

(I)

wherein HX represents at least one acid component, preferably hydrochloric, phosphoric acid, fumaric acid and L-tartaric acid. The invention also relates to the processes for the preparation thereof as well as said use thereof in phamaceutically acceptable compositions. Use of said crystalline forms of ruxolitinib and manufactured salts in the preparation of (3R)-3-cyclopentyl-3-[4-(7H- pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile in the free form or in the form of any phamaceutically acceptable salt thereof is also part of this invention.

Background Art

(3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile compound which is also known as ruxolitinib (CAS no.: 941678-49-5) has a selective inhibitor activity on the Janus Associated Kinase 1 (JAKl) and Janus Associated Kinase 2 (JAK2) enzymes. It is a drug indicated for the treatment of intermediate or high-risk myelofibrosis which is a type of bone marrow cancer. The enzymes Janus Associated Kinase 1 (JAKl) and Janus Associated Kinase 2 (JAK2) are non-receptor tyrosine kinases that mediate the signals via the JAK-STAT pathway. Cytokines play important roles in the control of the cell growth and the immune response. More specifically, Janus Associated Kinases are phosphorylate activated cytokine receptors recruiting STAT transcription factors which modulate gene transcription.

WO2007070514 describes protein kinase inhibitors with valuable pharmacological effect in the treatment of related diseases. One example of the compounds disclosed is (3 )-3-cyclopentyl-3-[4- (7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile. Preparation of the base is also described.

Salts of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile prepared with maleic acid, sulphuric acid and phosphoric acid are disclosed in WO2008157208. Following WO2010083283 discloses the process for the preparation of ruxolitinib phosphate with good yield and optical purity.

Many pharmaceutical solid compounds can exist in various crystalline forms regarded as polymorphs and hydrates/solvates having different crystal structures and hence different physico-chemical properties including melting point, solubility, dissolution rate and finally, bioavailability. In order to distinguish the distinct solid phases of a compound several solid state analytical techniques can be used, e.g. X-Ray Powder Diffraction, solid state NMR and Raman spectroscopy, thermoanalytical methods.

Discovery of new solid phases (polymorphs, solvates and hydrates) of an active pharmaceutical compound offers the opportunity to select the appropriate modification having desirable physico- chemical properties and processability and improve the characteristics of the pharmaceutical product. For this reason there is an explicit need for new solid forms (polymorphs, solvates, hydrates) of ruxolitinib and salts thereof escpecially in the crystalline form.

Disclosure of the Invention

The object of the present invention is to provide novel crystalline salts comprising (3R)-3-cyclopentyl- 3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile of Formula I and at least one acid component (HX) suitable for oral administration which meet the pharmaceutical requirements

(I)

wherein HX represents at least one acid component, preferably hydrochloric acid, phosphoric acid, fumaric acid, L-tartaric acid. In some embodiments of this invention, the solid forms are characterized by a variety of solid state analytical data, including for example X-ray powder diffraction pattern (XRPD) and differential scanning calorimetry (DSC) curve.

Provided is the Crystal modification 5 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile hydrochloric acid salt of Formula I, wherein X represents CI, having an X- ray powder diffraction pattern comprising characteristic peaks at about 3.7; 8.4; 14.3; 20.9 and 25.2 ± 0.2° 2-theta measured by CuKa radiation. In some embodiments the Crystal modification 5 is characterised by differential scanning calorimetry curve having a melting process with T_peak=90.9°C. In some embodiments the Crystal modification 5 is characterised by the thermal gravimetric curve having a 8.95% weigth loss in the range of 25°C to 80°C. It should be understood that relative intensities can vary depending on a number of factors, including sample preparation, mounting, and the instrument and analytical procedure and settings used to obtain the spectrum.

Provided is a process for the preparation of the Crystal modification 5, wherein the ruxolitinib hydrochloric acid salt is suspended in methanol-water mixture, appropriately ranging from 0 to 40%v/v of methanol in water and the suspension is stirred for about 72 hours. In some embodiments the process for the preparation of the Crystal modification 2 further comprises the steps of: a/ suspending ruxolitinib hydrochloric acid salt in water or methanol-water mixture, conveniently ranging from 0 to 40%v/v of methanol in water, conveniently at room temperature; b/ stirring the suspension of step a/ at room temperature for about 72 hours; c/ isolating the Crystal modification 5 of ruxolitinib hydrochloric acid salt and d/ optionally, drying of the product of step c/ under the laboratory conditions until the constant weight of the product is reached.

Provided is the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile phosphoric acid salt of Formula I, wherein X represents phosphate, having an X-ray powder diffraction pattern comprising characteristic peaks at about 3.9; 14.5; 15.9; 20.2 and 24.9 ± 0.2° 2-theta measured by CuKct radiation. In some embodiments the Crystal modification 1 is characterised by differential scanning calorimetry curve having a melting process with Τ_οη5ε1=141.3^ and T_Pea_k=191.5⁰C. In some embodiments the Crystal modification 1 is characterised by the thermal gravimetric curve having a 0.6% weigth loss in the range of 25°C to 190'C.

It should be understood that relative intensities can vary depending on a number of factors, including sample preparation and mounting, and the instrument and analytical procedure and settings used to obtain the spectrum. Another subject of present invention is a process for the preparation of the Crystal modification 1 wherein ruxolitinib free base is dissolved in a polar aprotic solvent and phosphoric acid is then added, preferably the polar aprotic solvent is at the temperature of 50°C, preferably followed by heating of the system to a temperature 50°C followed by the addition of the counterion, then kept at the temperature of 50°C for aditional 1 hour and finally cooled back to a temperature of 0-5°C. The polar aprotic solvent is selected from the group consisting of methyl-acetate , ethyl-acetate, butyl-acetate acetonitrile, acetone, methyl ethyl ketone or a mixture thereof, preferably the polar aprotic solvent is ethyl-acetate, more preferably it is ethyl-acetate at the temperature of 50°C.

In some embodiments the process for the preparation of the Crystal modification 1 further comprises the steps of: a/ dissolution of ruxolitinib free base in a polar aprotic solvent selected from the group consisting of: methyl-acetate , ethyl-acetate, butyl-acetate, acetonitrile, acetone, methyl ethyl ketone or a mixture thereof, more preferably in ethyl-acetate at the temperature of 50°C; b/ drop-wise addition of the phosphoric acid 85% aqueous solution; c/ stirring the solution of the step b/ at 50°C for aditional 1 hour; d/ cooling the solution of the step c/ to a temperature of 0-5°C while precipitation occurred; e/ keeping the suspension of the step d/ for 16 hours at a temperature of 0- 5^eC ; f/ isolating the ruxolitinib phosphoric acid salt in Crystal modification 1 and g/ optionally, drying of the product of step f/ under the laboratory conditions until the constant weight of the product is reached. Provided is the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile fumaric acid salt of Formula I, wherein X represents fumarate, having an X-ray powder diffraction pattern comprising characteristic peaks at about 5.6; 13.1; 18.1; 22.6; 23.5 and 25.5 ± 0.2° 2-theta measured by CuKct radiation. In some embodiments the Crystal modification 1 is characterised by differential scanning calorimetry curve having a melting process with Tpea_k=169.3^<>C. In some embodiments the Crystal modification 1 is characterised by the thermal gravimetric curve having a 0.6% weigth loss in the range of 25°C to 150°C.

It should be understood that relative intensities can vary depending on a number of factors, including sample preparation and mounting, and the instrument and analytical procedure and settings used to obtain the spectrum.

Another subject of present invention is a process for the preparation of the Crystal modification 1 wherein ruxolitinib free base is dissolved in a polar protic solvent and the solution of fumaric acid in a polar protic solvent is then added, preferably the polar protic solvent is at the temperature of 50°C, preferably followed by heating of the system to a temperature 50°C followed by the addition of the counterion, then kept at the temperature of 50°C for aditional 1 hour and finally cooled back to a temperature of 0-5°C. The polar protic solvent is selected from the group consisting of methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or a mixture thereof, preferably the polar protic solvent is ethanol, more preferably it is ethanol at the temperature of 50°C.

In some embodiments the process for the preparation of the Crystal modification 1 further comprises the steps of: a/ dissolution of ruxolitinib free base in a polar protic solvent selected from the group consisting of: methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or a mixture thereof, more preferably the suitable polar protic solvent is ethanol, more preferably it is ethanol at the temperature of 50°C; b/ drop-wise addition of the fumaric acid solution; c/ stirring the solution of the step b/ at 50°C for aditional 1 hour; d/ cooling the solution of the step c/ to a temperature of 0-5°C while precipitation occurred; e/ keeping the suspension of the step d/ for 16 hours at a temperature of 0-5°C ; f/ isolating the ruxolitinib fumaric acid salt in Crystal modification 1 and g/ optionally, drying of the product of step f/ under the laboratory conditions until the constant weight of the product is reached.

Provided is the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt of Formula I, wherein X represents L-tartrate, having an X-ray powder diffraction pattern comprising characteristic peaks at about 4.1; 8.2; 16.2; 20.9 and 25.5 ± 0.2° 2-theta measured by CuKa radiation. In some embodiments the Crystal modification 1 is characterised by differential scanning calorimetry curve having a melting process with T_onset=39. C and

In some embodiments the Crystal modification 1 is characterised by the thermal gravimetric curve having a 1.64% weigth loss in the range of 25°C to 110°C.

It should be understood that relative intensities can vary depending on a number of factors, including sample preparation and mounting, and the instrument and analytical procedure and settings used to obtain the spectrum.

Another subject of present invention is a process for the preparation of the Crystal modification 1 wherein ruxolitinib free base is dissolved in a polar protic solvent and the solution of L-tartaric acid in a polar protic solvent is then added, preferably the polar protic solvent is at the temperature of 50°C, preferably followed by heating of the system to a temperature 50°C followed by the addition of the counterion, then kept at the temperature of 50°C for aditional 1 hour and finally cooled back to a temperature of 0-5°C. The polar protic solvent is selected from the group consisting of methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or a mixture thereof, preferably the polar protic solvent is ethanol, more preferably it is ethanol at the temperature of 50°C.

In some embodiments the process for the preparation of the Crystal modification 1 further comprises the steps of: a/ dissolution of ruxolitinib free base in a polar protic solvent selected from the group consisting of: methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or a mixture thereof, more preferably the suitable polar protic solvent is ethanol, more preferably it is ethanol at the temperature of 50°C; b/ drop-wise addition of the L-tartaric acid acid solution; c/ stirring the solution of the step b/ at 50°C for aditional 1 hour; d/ cooling the solution of the step c/ to a temperature of 0- 5°C while precipitation occurred; e/ keeping the suspension of the step d/ for 16 hours at a temperature of 0-5°C ; f/ isolating the ruxolitinib L-tartarric acid salt in Crystal modification 1 and g/ optionally, drying of the product of step f/ under the laboratory conditions until the constant weight of the product is reached.

Provided is the Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt of Formula I, wherein X represents L-tartrate, having an X-ray powder diffraction pattern comprising characteristic peaks at about 6.3; 9.1; 13.3; 16.3; 18.7; 19.4 and 22.8 ± 0.2° 2-theta measured by CuKoc radiation. In some embodiments the Crystal modification 1 is characterised by differential scanning calorimetry curve having a melting process with

In some embodiments the Crystal modification 1 is characterised by the thermal gravimetric curve having a 3.42% weigth loss in the range of 25°C to 97°C.

It should be understood that relative intensities can vary depending on a number of factors, including sample preparation and mounting, and the instrument and analytical procedure and settings used to obtain the spectrum. Another subject of present invention is a process for the preparation of the Crystal modification 2 wherein ruxolitinib free base is dissolved in a polar aprotic solvent and the solution of L-tartaric acid is then added, preferably the polar aprotic solvent is at the temperature of 50°C, preferably followed by heating of the system to a temperature 50°C followed by the addition of the counterion, then kept at the temperature of 50°C for aditional 1 hour and finally cooled back to a temperature of 0-5"C. The polar aprotic solvent is selected from the group consisting of methyl-acetate , ethyl-acetate, butyl- acetate, acetonitrile, acetone, methyl ethyl ketone or a mixture thereof, preferably the polar aprotic solvent is ethyl-acetate, more preferably it is acetonitrile at the temperature of 50°C.

In some embodiments the process for the preparation of the Crystal modification 2 further comprises the steps of: a/ dissolution of ruxolitinib free base in a polar aprotic solvent selected from the group consisting of: methyl-acetate , ethyl-acetate, butyl-acetate acetonitrile, acetone, methyl ethyl ketone or a mixture thereof, more preferably in acetonitrile, more preferably in acetonitrile at the temperature of 50°C; b/ drop-wise addition of the L-tartaric acid acid solution; c/ stirring the solution Of the step b/ at 50°C for aditional 1 hour; d/ cooling the solution of the step c/ to a temperature of 0- 5°C while precipitation occurred; e/ keeping the suspension of the step d/ for 16 hours at a temperature of 0-5°C ; f/ isolating the ruxolitinib L-tartarric acid salt in Crystal modification 1 and g/ optionally, drying of the product of step f/ under the laboratory conditions until the constant weight of the product is reached.

Brief description of the Drawings the figures depict the following spectra, patterns and curves the various solid phases prepared according to the present invention.

Figure 1 is an XRPD pattern of the Crystal modification 5 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile hydrochloric acid salt; Figure 2 is an FTIR spectra of the Crystal modification 5 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- ^] yrimidin-4-yl)pyrazol-l-yl]propanenitrile hydrochloric acid salt;

Figure 3 is a Raman spectra of the Crystal modification 5 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile hydrochloric acid salt;

Figure 4 is a DSC curve of the Crystal modification 5 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile hydrochloric acid salt; Figure 5 is a TGA curve of the Crystal modification 5 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile hydrochloric acid salt;

Figure 6 is an XRPD pattern of the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile phosphoric acid salt;

Figure 7 is an FTIR spectra of the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile phosphoric acid salt;

Figure 8 is a Raman spectra of the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile phosphoric acid salt;

Figure 9 is a DSC curve of the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile phosphoric acid salt;

Figure 10 is a TGA curve of the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile phosphoric acid salt;

Figure 11 is a ¹H-NMR spectra of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile and phosphoric acid salt prepared according to Example 2;

Figure 12 is an XRPD pattern of the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile fumaric acid salt;

Figure 13 is am FTIR spectra of the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile fumaric acid salt;

Figure 14 is a Raman spectra of the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile fumaric acid salt;

Figure 15 is a DSC curve of the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile fumaric acid salt;

Figure 16 is a TGA curve of the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile fumaric acid salt;

Figure 17 is a ^-NMR spectra of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile and fumaric acid salt prepared according to Example 4;

Figure 18 is an XRPD pattern of the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt;

Figure 19 is an FTIR spectra of the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt; Figure 20 is a Raman spectra of the Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt;

Figure 21 is a DSC curve of the Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt; Figure 22 is a TGA curve of the Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt;

Figure 23 is a ¹H-NMR spectra of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile and L-tartaric acid salt prepared according to Example 5;

Figure 24 is an XRPD pattern of the Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt;

Figure 25 is an FTIR spectra of the Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt;

Figure 26 is a Raman spectra of the Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt; Figure 27 is a DSC curve of the Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt;

Figure 28 is a TGA curve of the Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt;

Figure 29 is a 1H-NMR spectra of the (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol- l-yl]propanenitrile and L-tartaric acid salt prepared according to Example 6

Detailed description of the Invention

The following description is presented to enable a person of ordinary skill in the art to make and use the various embodiments. Description of specific devices, techniques, and applications are provided only as examples. Various modifications to the examples described herein will be readily apparent to those of ordinary skill in the art, and the general principles described herein may be applied to other examples and applications without departing from the spirit and scope of the various embodiments. Therefore, the various embodiments are not intended to be limited to the examples described herein and shown, but are to be accorded the scope consistent with the claims. The aim of the present invention is to provide novel crystalline forms of (3R)-3-cyclopentyl-3-[4-(7H- pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile salts of Formula I,

(I) wherein HX represents at least one acid component, preferably hydrochloric acid, phosphoric acid, fumaric acid and L-tartaric acid, with advantegous properties for pharmaceutical use regarding the physico-chemical properties and can be produced in a reproducible manner even in industrial scale. The invention also relates to the processes for the preparation thereof as well as said use thereof in phamaceutically acceptable compositions. Use of said crystalline forms of ruxolitinib and manufactured salts in the preparation of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile in the free form or in the form of any phamaceutically acceptable salt thereof is also part of this invention.

Variations in the crystal structure of ruxolitinib salts may affect the dissolution rate (which may affect bioavailability etc.), manufacturability (e.g., ease of handling, ability to consistently prepare doses of ; i different strength) and stability (e.g., thermal stability, shelf life, etc.) of a pharmaceutical drug u product, particularly when formulated in a solid oral dosage form (e.g., in a form of a tablet). The therapeutic use and manifacturing of ruxolitinib involves the development of a new solid form of ruxolitinib salts that is more bioavailable and stable.

The term„form" of ruxolitinib, as used in this document, is synonymous to terms„solid state form, solid phase variant" of ruxolitinib and includes crystalline modificatiohs, hydrates and solvates of ruxolitinib. The term„crystal modification" of ruxolitinib, as used in this document, is synonymous to commonly used expressions„ polymorphic form" or„crystalline form" of ruxolitinib.

The use of the term "about" includes and describes the value or parameter per se. For example, "about x" includes and describes "x" per se. In some embodiments, the term "about" when used in association with a measurement, or used to modify a value, a unit, a constant, or a range of values, refers to variations of +/- 20 percent, preferably +/- 10 percent and even more preferably +/- 5 percent.

The term "substantially" or "substantially free/pure" with respect to a particular solid form of a compound means that the polymorphic form contains about less than 30 percent, about less than 20 percent, about less than 15 percent, about less than 10 percent, about less than 5 percent, or about less than 1 percent by weight of impurities. In other embodiments, "substantially" or "substantially free/pure" refers to a substance free of impurities. Impurities may, for example, include by-products or left over reagents from chemical reactions, contaminants, degradation products, other polymorphic forms, water and solvents. It has now been surprisingly found that the above-mentioned crystalline salts of (3R)-3-cyclopentyl-3- [4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l-yl] with at least one acid component HX, wherein HX is preferably hydrochloric acid, phosphoric acid, fumaric acid and L-tartaric acid can be prepared and have not been described in the literature yet and no solid state analytical data (X-Ray Powder Diffraction patterns, Single-Crystal X-Ray Diffraction data etc.) serving to characterize the crystalline phases have been provided.

The Crystal modification 5 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile hydrochloric acid salt according to the invention has the characteristic XRPD pattern as shown in Figure 1. XRPD pattern was recorded on an X-Ray Powder Diffractometer (X'PERT PRO PD PANalytical). The Crystal modification 5 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin- 4-yl)pyrazol-l-yl]propanenitrile hydrochloric acid salt exhibits the following diffraction peaks in XRPD pattern, see Table 1 below:

Pos.

[°2Th.] d [A] Rel. Int. [%]

3.68 23.979 100.0

7.50 11.782 26.2 8.38 10.543 82.0

8.93 9.894 23.1

9.81 9.005 35.0

10.21 8.659 12.3

14.33 6.178 42.5

14.68 6.029 18.7

15.15 5.843 32.0

16.65 5.321 20.3

16.95 5.226 22.9

17.33 5.113 10.4

17.77 4.988 17.6

18.77 4.723 30.8

19.48 4.554 19.0

20.93 4.242 53.8

21.80 4.078 6.5

22.22 3.998 8.1

22.81 3.896 22.2

23.38 3.802 23.4

23.71 3.749 31.5

24.04 3.700 16.7

24.47 3.635 12.1

25.19 3.533 59.8

25.63 3.472 24.4

26.10 3.412 40.2

26.80 3.324 25.7

28.12 3.171 16.3

29.04 3.072 18.9

Table 1

The Crystal modification 5 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile hydrochloric acid salt can be characterized by FTIR and Raman spectroscopy. Figure 2 shows the FTIR spectrum (Nicolet Thermo 6700c ) comprising characteristic peaks at 3395, 3094, 2954, 2868, 2247, 1619, 1587, 1347, 816 and 742 cm ¹ wavenumbers. The Crystal modification 5 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile hydrochloric acid salt is characterised by a Raman spectrum (Bruker RFS 100/S) comprising characteristic peaks at 3115, 3068, 2964, 2951, 2872, 2247, 1624, 1549, 1315 and 593 cm^'1 wavenumbers, shown in Figure 3.

The Crystal modification 5 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile hydrochloric acid salt can be further described by thermal analytical methods. Figure 4 shows the DSC (Mettler-Toledo 822e DSC) and Figure 5 shows the TGA (NETZSCH TG 209 thermogravimetric analyser) curves measured in the range of 25°C to 350°C and 25°C to 350°C, respectively. The Crystal modification 5 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile hydrochloric acid salt shows a 8.95% weigth loss in the range of 25°C to 80°C. The DSC measurement gives a melting process with T_peak=90.9°C.

One subject of the invention is process for preparation of the Crystal modification 5. In this process, ruxolitinib hydrochloric acid salt is dissolved in a suitable organic solvent at room temperature. The suitable organic solvent is preferably a polar protic solvent or this organic solvent-water mixture, more preferably the polar protic solvent is selected from the group consisting of: methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or an organic solvent-water mixture thereof, more preferably in methanol-water mixture, even more preferably methanol-water mixture ranging from 0 to 40%v/v of water in methanol at room temperature.

The Crystal modification 5 of ruxolitinib hydrochloric acid salt can be prepared by a process comprising the steps of: a/ suspending ruxolitinib hydrochloric acid salt in in water or methanol-water mixture, preferably angirrg from 0 to 40%v/v of methanol in water at room temperature; b/ stirring the suspension of step a/ at room temperature for 72 hours; c/ isolating the Crystal modification 5 of ruxolitinib hydrochloric acid salt; d/ optionally, drying of the product of step c/ under the laboratory conditions until the constant weight of the product is reached.

i The suitable organic solvent is preferably a polar protic solvent, more preferably the polar prptic solvent or an organic solvent-water mixture thereof is selected from the group consisting of: methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or a mixture thereof, more preferably in methanol-water mixture, even more preferably methanol-water mixture ranging from 0 to 40%v/v of water in methanol at room temperature. The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile phosphoric acid salt according to the invention has the characteristic XRPD pattern as shown in Figure 6. XRPD pattern was recorded on an X-Ray Powder Diffractometer (X'PERT PRO MPD PANalytical). The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrroio[2,3-d]pyrimidin- 4-yl)pyrazol-l-yl]propanenitriie phosphoric acid salt exhibits the following diffraction peaks in XRPD pattern, see Table 2 below:

Pos.

[°2Th.] d [A] Rel. Int. [%]

3.95 22.370 100.0

4.79 18.430 36.8

7.52 11.751 1.6

7.90 11.120 1.1

8.27 10.682 1.9

9.69 9.122 1.9

12.05 7.338 1.4

12.46 7.099 1.5

12.92 6.849 4.9

13.81 6.410 2.8

14.09 6.281 2.5

14.49 6.110 13.7

14.75 6.002 5.1

15.89 5.575 17.0

16.46 5.381 2.5

16.68 5.312 3.5

17.33 5.113 4.9

17.64 5.024 3.8

18.70 4.743 8.7

19.63 4.519 11.3

20.16 4.402 13.9

20.80 4.268 8.1

21.68 4.097 11.0 22.71 3.912 4.5

23.13 3.843 5.6

23.74 3.745 4.9

24.86 3.578 12.4

25.21 3.529 10.7

26.29 3.387 9.9

Table 2

The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yljpropanenitrile phosphoric acid salt can be characterized by FTIR and Raman spectroscopy. Figure 7 shows the FTIR spectrum (Nicolet Thermo 6700c ) comprising characteristic peaks at 3111, 2949, 2871, 2251, 1621, 1599, 1435, 1078, 815 and 732 cm ¹ wavenumbers. The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile phosphoric acid salt is characterised by a Raman spectrum (Bruker RFS 100/S) comprising characteristic peaks at 3146, 3126, 2967, 2931, 2873, 2252, 1624, 1496, 1306 and 818 cm^"1 wavenumbers, shown in Figure 8. The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile phosphoric acid salt can be further described by thermal analytical methods. Figure 9 shows the DSC (Mettler-Toledo 822e DSC) and Figure 10 shows the TGA (NETZSCH TG 209 thermogravimetric analyser) curves measured in the range of 25°C to 350°C and 25°C to 350°C, respectively. The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile phosphoric acid salt shows a 0.6% weigth loss in the range of 25°C to 190°C. The DSC measurement gives a melting process with

and T_peak=191.5°C.

In one of the aspects of the invention, process for preparation of the Crystal modification 1 is provided. In this process, ruxolitinib free base is dissolved in a suitable organic solvent by heating of the system to a temperature 50°C. The phosphoric acid is added to the solution. The suitable organic solvent is preferably a polar aprotic solvent, more preferably the polar aprotic solvent thereof is selected from the group consisting of: methyl-acetate , ethyl-acetate, butyl-acetate acetonitrile, acetone, methyl ethyl ketone or a mixture thereof, preferably the polar aprotic solvent is ethyl- acetate, more preferably it is ethyl-acetate at the temperature of 50°C.

The process of preparation of the Crystal modification 1 of ruxolitinib phosphoric acid salt thus comprises the steps of: a/ dissolution of ruxolitinib free base in a suitable organic solvent; b/ drop-wise addition of the phosphoric acid 85% aqueous solution while precipitation occurred; c/ stirring the suspension of the step b/ at 50"C for aditional 1 hour; d/ cooling the suspension of the step c/ to a temperature of 0-5°C; e/ keeping the suspension of the step d/ for 16 hours at a temperature of 0-5"C ; f/ isolating the ruxolitinib phosphoric acid salt in Crystal modification 1; gl optionally, drying of the product of step f/ under the laboratory conditions until the constant weight of the product is reached.

The suitable organic solvent is preferably a polar aprotic solvent, more preferably the polar aprotic solvent thereof is selected from the group consisting of: methyl-acetate , ethyl-acetate, butyl-acetate, , acetonitrile, acetone, methyl ethyl ketone or a mixture thereof, preferably the polar aprotic solvent is ethyl-acetate, more preferably it is ethyl-acetate at the temperature of 50°C.

Another process for the preparation of Crystal modification 1 of ruxolitinib phosphoric acid salt comprises the steps of: a/ suspending ruxolitinib phosphoric acid salt in ethyl-acetate at room temperature; b/ stirring the suspension of step a/ at room temperature for 72 hours; c/ isolating the Crystal modification 1 of ruxolitinib phosphoric acid salt;

. d/ optionally, drying of the product of step c/ under the laboratory conditions until the constant weight of the product is reached.

The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- . yllpropanenitrile fumaric acid salt according to the invention has the characteristic XRPD pattern as shpwn in Figure 12. XRPD pattern was recorded on an X-Ray Powder Diffractometer (X'PERT PRO MPD PANalytical). The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin- 4-yl)pyrazol-l-yl]propanenitrile fumaric acid salt exhibits the following diffraction peaks in XRPD ^Γ;/' pattern, see Table 3, below: Pos. Rel. Int.

[°2Th.] d [A] [%]

5.63 15.677 100.0

6.48 13.631 82.6

10.23 8.643 11.3

10.48 8.437 15.0

13.05 6.781 24.7

14.88 5.949 16.5

15.23 5.815 12.1

15.49 5.714 22.2

16.26 5.448 26.1

16.65 5.320 9.8

17.88 4.958 29.0

18.11 4.894 48.2

19.35 4.584 19.5

19.89 4.461 32.6

20.40 4.350 6.3

21.00 4.227 5.9

22.07 4.025 27.3

22.66 3.921 46.3

23.10 3.848 14.5

23.55 3.775 45.7

23.87 3.725 14.2

24.30 3.660 17.0

24.57 3.620 11.3

24.87 3.577 8.3

25.54 3.484 46.6

26.16 3.404 9.4

26.52 3.358 18.5

28.54 3.125 23.5

Table 3

The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yllpropanenitrile fumaric acid salt can be characterized by FTIR and Raman spectroscopy. Figure 13 shows the FTI spectrum (Nicolet Thermo 6700c ) comprising characteristic peaks at 3201, 3132, 2957, 2266, 1696, 1592, 1346, 831, 737 and 608 cm ¹ wavenumbers. The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile fumaric acid salt is characterised by a Raman spectrum (Bruker RFS 100/S) comprising characteristic peaks at 3150, 3131, 3074, 2940, 2876, 2266, 1702, 1594, 1350 and 829 cm^"1 wavenumbers, shown in Figure 14.

The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile fumaric acid salt can be further described by thermal analytical methods. Figure 15 shows the DSC (Mettler-Toledo 822e DSC) and Figure 16 shows the TGA (NETZSCH TG 209 thermogravimetric analyser) curves measured in the range of 25°C to 350°C and 25°C to 350°C, respectively. The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile fumaric acid salt shows a 0.6% weigth loss in the range of 25°C to 150°C. The DSC measurement gives a melting process with T_peak=169.3°C.

In one of the aspects of the invention, process for preparation of the Crystal modification 1 is provided. In this process, ruxolitinib free base is dissolved in a suitable organic solvent by heating of the system to a temperature 50°C. The fumaric acid is added to the solution. The suitable organic solvent is preferably a polar protic solvent selected from the group consisting of: methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or a mixture thereof, more preferably in ethanol, even more preferably ethanol at the temperature of 50°C.

The process of preparation of the Crystal modification 1 of ruxolitinib fumaric acid salt thus comprises the steps of: a/ dissolution of ruxolitinib free base in a suitable organic solvent;

b/ drop-wise addition of the solution of fumaric acid solution in a suitable organic solvent while precipitation occurred; c/ stirring the suspension of the step b/ at 50°C for aditional 1 hour; d/ cooling the suspension of the step c/ to a temperature of 0-5°C;

e/ keeping the suspension of the step d/ for 16 hours at a temperature of 0-5°C ; f/ isolating the ruxolitinib fumaric acid salt in Crystal modification 1;

gl optionally, drying of the product of step f/ under the laboratory conditions until the constant weight of the product is reached. The suitable organic solvent is preferably a polar protic solvent selected from the group consisting of: methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or a mixture thereof, more preferably in ethanol, even more preferably ethanol at the temperature of 50°C.

Another process for the preparation of the Crystal modification 1 of ruxoiitinib fumaric acid salt comprises the steps of: a/ suspending ruxoiitinib fumaric acid salt in ethanol at room temperature; b/ stirring the suspension of step a/ at room temperature for 1 week;

c/ isolating the Crystal modification 1 of ruxoiitinib fumaric acid salt; d/ optionally, drying of the product of step c/ under the laboratory conditions until the constant weight of the product is reached.

i- The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yTjpropanenitrile L-tartaric acid salt according to the invention has the characteristic XRPD pattern as

, shp.wn in; Figure 18. XRPD pattern was recorded on an X-Ray Powder Diffractometer (X'PERT PRO MPD PANalytical). The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin- 4-yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt exhibits the following diffraction peaks in XRPD pattern, see Table 4, below:

Pos. Rel. Int.

[°2Th.] d [A] [%}

4.099 21.5389 100

8.147 10.84423 20.31

11.329 7.80446 3.17

11.715 7.54802 2.44

14.096 6.27776 12.68

16.214 5.46214 20.01

17.28 5.1276 7.74

17.64 5.02318 5.91

18.712 4.73824 14.9

19.352 4.58294 11.51

20.873 4.25234 14.23 21.672 4.09746 6.26

22.705 3.91321 9.18

23.53 3.77794 6.29

24.687 3.6033 11.87

25.521 3.48748 12.22

26.13 3.40762 10.46

27.585 3.23108 8.05

Table 4

The Crystal modification 1 of (3 )-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile L-tartaric acid salt can be characterized by FTIR and Raman spectroscopy. Figure 19 shows the FTIR spectrum (Nicolet Thermo 6700c ) comprising characteristic peaks at 3390, 3178, 3124, 2951, 2867, 2257, 1625, 1339, 1074 and 763 cm^"1 wavenumbers. The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt is characterised by a Raman spectrum (Bruker RFS 100/S) comprising characteristic peaks at 3122, 2961, 2926, 2872, 2259, 1628, 1356, 1241, 776 and 591 cm ¹ wavenumbers, shown in Figure 20.

The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile L-tartaric acid salt can be further described by thermal analytical methods. Figure 21 shows the DSC (Mettler-Toledo 822e DSC) and Figure 22 shows the TGA (NETZSCH TG 209 thermogravimetric analyser) curves measured in the range of 25°C to 350°C and 25°C to 350°C, respectively. The Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt shows a 1.64% weigth loss in the range of 25°C to 110°C. The DSC measurement gives a melting process with

116.4°C.

In one of the aspects of the invention, process for preparation of the Crystal modification 1 is provided. In this process, ruxoiitinib free base is dissolved in a suitable organic solvent by heating of the system to a temperature 50°C. The L-tartaric acid is added to the solution. The suitable organic solvent is preferably a polar protic solvent, more preferably the polar protic solvent is selected from the group consisting of: methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or a mixture thereof, even more preferably the suitable organic solvent is ethanol, even more preferably the suitable organic solvent is ethanol at the temperature of 50°C. The Crystal modification 1 of ruxolitinib L-tartaric acid salt can be prepared by a process comprising the steps of: a/ dissolution of ruxolitinib free base in a suitable organic solvent; b/ drop-wise addition of the solution of L-tartaric acid solution in a suitable organic solvent; 5 c/ stirring the solution of the step b/ at 50°C for aditional 1 hour; d/ cooling the solution of the step c/ to room temperature; e/ keeping the solution of the step d/ for 16 hours at room temperature while precipitation occurred; f/ isolating the ruxolitinib L-tartaric acid salt in Crystal modification 1; g/ optionally, drying of the product of step f/ under the laboratory conditions until the constant 10 weight of the product is reached.

The suitable organic solvent is preferably a polar protic solvent, more preferably the polar protic solvent is selected from the group consisting of: methanol, ethanol, 1-propanol, 2-propanol, 1- butanol, 2-butanol or a mixture thereof, even more preferably the suitable organic solvent is ethanol, even more preferably the suitable organic solvent is ethanol at the temperature of 50°C.

15

The Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile L-tartaric acid salt according to the invention has the characteristic XRPD pattern as shown in Figure 24. XRPD pattern was recorded on an X-Ray Powder Diffractometer (X'PERT PRO , MPD PANalytical). The Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin- ! 20 4-y|)pyrazol-l-yl]propanenitrile L-tartaric acid salt exhibits the following diffraction peaks in XRPD pattern, see Table 5, below:

Pos. Rel. Int.

[°2Th.] d [A] [%]

6.34 13.940 100.0

8.12 10.879 8.1

9.06 9.749 34.9

9.64 9.168 5.2

10.89 8.118 12.0

11.70 7.560 4.0 13.27 6.665 52.3

14.18 6.239 6.3

14.52 6.096 8.9

15.78 5.613 6.9

16.28 5.440 34.0

16.76 5.284 11.8

16.95 5.227 10.4

17.86 4.963 27.6

18.20 4.872 26.6

18.70 4.741 26.4

19.00 4.667 16.5

19.39 4.573 30.9

19.82 4.476 13.7

21.32 4.164 4.5

21.92 4.052 6.2

22.76 3.904 13.2

23.27 3.820 6.7

23.89 3.722 8.6

24.60 3.615 8.6

25.06 3.550 11.8

25.33 3.513 14.3

25.72 3.461 14.9

26.53 3.357 9.4

27.16 3.281 7.2

28.75 3.103 6.9

29.47 3.028 5.0

30.11 2.966 4.5

Table 5

The Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile L-tartaric acid salt can be characterized by FTIR and Raman spectroscopy. Figure 25 shows the FTIR spectrum (Nicolet Thermo 6700c ) comprising characteristic peaks at 3192, 3116, 2948, 2864, 2252, 1721, 1596, 1346, 1266 and 816 cm^"1 wavenumbers. The Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt is characterised by a Raman spectrum (Bruker RFS 100/S) comprising characteristic peaks at 3148, 2956, 2933, 2869, 2253, 1619, 1582, 1350, 1144 and 819 cm^"1 wavenumbers, shown in Figure 26.

The Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- yl]propanenitrile L-tartaric acid salt can be further described by thermal analytical methods. Figure 27 shows the DSC (Mettler-Toledo 822e DSC) and Figure 28 shows the TGA (NETZSCH TG 209 thermogravimetric analyser) curves measured in the range of 25°C to 350°C and 25°C to 350°C, respectively. The Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt shows a 3.42% weigth loss in the range of 25°C to 97°C. The DSC measurement gives a melting process with

In one of the aspects of the invention, process for preparation of the Crystal modification 2 is provided. In this process, ruxolitinib free base is suspended in a suitable organic solvent by heating of the system to a temperature 50°C. The L-tartaric acid is added to the solution. The suitable organic solvent is preferably a polar aprotic solvent, more preferably the polar aprotic solvent is selected from the group consisting of: acetone, acetonitrile, methyl ethyl ketone, ethyl acetate, butyl acetate, tetrahydrofuran or a mixture thereof, even more preferably the suitable organic solvent is acetonitrile, even more preferably the suitable organic solvent is acetonitrile at the temperature of 50°C.

The Crystal modification 2 of ruxolitinib L-tartaric acid salt can be prepared by a process comprising the steps of: a/ suspending of ruxolitinib free base in a suitable organic solvent; b/ drop-wise addition of the solution of L-tartaric acid solution in a suitable organic solvent;

c/ stirring the solution of the step b/ at 50°C for aditional 1 hour while precipitation occurred; d/ cooling the suspension of the step c/ to room temperature; e/ keeping the suspension of the step d/ for 16 hours at room temperature; f/ isolating the ruxolitinib L-tartaric acid salt in Crystal modification 2;

gl optionally, drying of the product of step f/ under the laboratory conditions until the constant weight of the product is reached.

The suitable organic solvent is preferably a polar aprotic solvent, more preferably the polar aprotic solvent is selected from the group consisting of: acetone, acetonitrile, methyl ethyl ketone, ethyl acetate, butyl acetate, tetrahydrofuran or a mixture thereof, even more preferably the suitable organic solvent is acetonitrile, even more preferably the suitable organic solvent is acetonitrile at the temperature of 50°C.

The term„room temperature" is defined as a temperature between 15°C and 29°C for the purpose of this document; preferably it is between 20-23°C. The term„drying under the laboratory conditions", as used in this patent application, means drying at room temperature and relative humidity 20-60%.

Analysis - XRPD (X-Ray Powder Diffractometry)

Diffractograms were obtained with laboratory X'PERT PRO MPD PANalytical diffractometer, used radiation CuKa (λ = 1.542 A).

Generator settings: excitation voltage 45 kV

- anodic current 40 mA

Scan description: - scan type - gonio

measurement range 2 - 40° 2Θ

- step size 0.01° 2Θ

step time: 0.5 s

Samples were measured as received on Si plate (zero background holder). Incident beam optics: programmable divergence slits (irradiated length 10 mm). 10 mm mask. 1/49 anti-scatter fixed slit, 0.02 rad Soller slits.

Diffracted beam optics: X'Celerator detector, scanning mode, active length 2.122S. 0.02 rad Soller slits, anti-scatter slit 5.0 mm Ni filter.

Analysis - FTIR (Fourier-Transformed Infra-Red) spectroscopy

FTIR spectra were recorded by Nicolet Thermo 6700 spectrometer.

General settings:

Number of sample scans: 45

Number of background scans: 45 Resolution: 4.000

Sample gain: 4.0

Optical velocity: 0.6329

Aperture: 100.00

Analysis - Raman spectroscopy

FTIR spectra were recorded by FT-Raman Bruker RFS 100/S Spectrometer General settings:

Excitation source: Nd-YAG laser (1064 nm) Applied spectral domain: 4000-200 cm^"1

Applied laser power: 250 mW

Detector: liquid nitrogen cooled Ge-diode detector (D418-T) Resolution: 4 cm^"1 Number of accumulations: 128 Scattering geometry: 180° (back scattering)

Aperture: 3.5 mm

Analysis - DSC (Differential Scanning Calorimetry)

DSC measurements were performed using a ettler-Toledo 822e DSC. Samples were placed into standard aluminum pans (40 μί) sealed with a pierced lid. The sample cell was heated under a nitrogen purge at a rate of 10°C/min from 25°C up to a final temperature of 300°C with 50 mL/min nitrogen purge. The temperatures specified in relation to DSC analyses are the temperatures of the peak maxima (T_peak) and onset temperature (T_onset) of peaks for the crystalline form. The enthalpy is given in J/g. The weight sample was about 2.5-3 mg. Analysis - TGA (ThermoGravimetric Analysis)

TGA analyses were performed using a NETZSCH TG 209 thermogravimetric analyser (NETZSCH- Geratebau GmbH, Germany).

Each sample was placed in an aluminum sample pan and inserted into the TG furnace. The furnace was heated under nitrogen purge at a rate of 10°C/min from 25°C up to a final temperature of 300°C.

The weight sample was about 5-15 mg.

Examples

The following examples are intended to further illustrate the present invention without limiting its scope.

Example 1

Preparation of Crystal modification 5 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile hydrochloric acid salt acid salt 200 mg (0.516 mmol) of ruxolitinib hydrochloric acid salt was suspended in 0.2 mL of 90-10 v/v% methanol-water mixture at room temperature.

The suspension was kept in closed vessel at room temperature applying a continuous stirring for 72 hours.

The solid obtained was collected by filtration and dried by vacuum suction at laboratory condition. XRPD pattern was measured (Figure 1) and showed that the compound is in a crystalline state that was designated as Crystal modification 5 of ruxolitinib hydrochloric acid salt.

Example 2

Preparation of Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile phosphoric acid salt

2 g (6.528 mmol) of ruxolitinib free base was suspended in 10 mL of ethyl-acetate by heating to 50°C applying a continuous stirring.

0.4 mL of phosphoric acid (6.528 mmol; 85% aqueous solution) was drop-wise added to the suspension of ruxolitinib at 50°C, while continuously stirred. The suspension was stirred at this temperature for 1 hour then placed onto ice to cool. The solid obtained was filtered off and dried at laboratory condition.

Product: 2.506 g (6.19 mmol) off-white crystalline solid

Yield: 95%

5 HPLC: 99.8%

XRPD pattern was measured (Figure 6) and showed that the compound is in a crystalline state that was designated as Crystal modification 1 of ruxolitinib phosphoric acid salt.

¹H-NMR spectrum was measured (Figure 11) and showed that the compound confirms the structure with an API : phosphoric acid stoichiometry of 1:1.

Example 3

Preparation of Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile phosphoric acid salt

^ 200 mg (0.495 mmol) of ruxolitinib phosphoric acid salt was suspended in 0.5 mL of ethyl-acetate at 15 room temperature.

The suspension was kept in closed vessel at room temperature applying a continuous stirring for 1 week.

The solid obtained was collected by filtration and dried by vacuum suction at laboratory condition.

Q XRPD pattern was measured (Figure 6) and showed that the compound is in a crystalline state that 20 was designated as Crystal modification 1 of ruxolitinib phosphoric acid salt.

Example 4

Preparation of Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile fumaric acid salt

25 2 g (6.528 mmol) of ruxolitinib free base was dissolved in 5 mL of ethanol at 50°C applying a continuous stirring.

The solution of 0.8 g of fumaric acid dissolved in 2 mL ethanol was drop-wise added to the solution of ruxolitinib at 50°C, while continuously stirred. Precipitation occurred. The suspension was further stirred at 50°C for additional 1 hour and then cooled back to room temperature. The suspension was further stirred at laboratory conditions overnight.

The solid precipitated is collected by filtration and dried at laboratory condition.

Product: 1.96 g (5.378 mmol) off-white crystalline solid Yield: 89%

HPLC: 97.7%

XRPD pattern was measured (Figure 12) and showed that the compound is in a crystalline state that was designated as Crystal modification 1 of ruxolitinib fumaric acid salt.

¹H-NMR spectrum was measured (Figure 17) and showed that the compound confirms the structure with an API : fumaric acid stoichiometry of 2:1

Example 5

Preparation of Crystal modification 1 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt 2 g (6.528 mmol) of ruxolitinib is dissolved in 8 mL of ethanol at 50°C applying a continuous stirring.

The solution of 1.0 g L-tartaric acid (6.666 mmol) dissolved in 2 mL ethanol is drop-wise added to the solution of ruxolitinib at 50°C, while continuously stirred.

The solution is further stirred at 50°C for additional 1 hour, while precipitation occurred.

The suspension formed was cooled back to room temperature and stirred overnight at that temperature.

The solid precipitated is collected by filtration and dried at laboratory condition Product: 2.14 g (4.7 mmol) off-white crystalline solid

Yield: 72% HPLC: 99.1% XRPD pattern was measured (Figure 18) and showed that the compound is in a crystalline state that was designated as Crystal modification 1 of ruxolitinib L-tartaric acid salt.

¹H-NMR spectrum was measured (Figure 23) and showed that the compound confirms the structure with an API: L-tartaric acid stoichiometry of 1:1 Example 6

Preparation of Crystal modification 2 of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4- yl)pyrazol-l-yl]propanenitrile L-tartaric acid salt

1 g (3.264 mmol) of ruxolitinib is suspended in 8 mL of acetonitrile at 50°C applying a continuous stirring.

The solution of 0.54 g of L-tartaric acid (3.6 mmol)in 2 mL ethyl-acetate is drop-wise added to the suspension of ruxolitinib at 50°C, while continuously stirred, forming a clear solution.

The solution is further stirred at 50°C for additional 1 hour, while precipitation occurred.

The suspension formed was cooled back to room temperature and stirred overnight at that temperature.

The solid precipitated is collected by filtration and dried at laboratory condition Product: 1.18 g (2.587 mmol) off-white crystalline solid Yield: 79% HPLC: 99.4% XRPD pattern was measured (Figure 24) and showed that the compound is in a crystalline state that was designated as Crystal modification 5 of ruxolitinib L-tartaric acid salt.

¹H-NMR spectrum was measured (Figure 29) and showed that the compound confirms the structure with an API: L-tartaric acid stoichiometry of 1:1.5

Claims

1. A salt comprising (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-l- -yl]propanenitrile of Formula I and at least one acid component (HX), characterized in that the acid addition salt is in a crystalline form.

H-X

2. The crystalline salt in accordance with claim 1, characterized in that the acid component (HX) is a hydrochloric acid, phosphoric acid, fumaric acid or L-tartaric acid.

3. A Crystal modification 5 of the crystalline salt in accordance with claims 1 and 2, wherein the acid component (HX) is a hydrochloric acid, showing in an X-ray powder diffraction (XRPD) pattern characteristic peaks at about 3.7; 8.4; 14.3; 20.9 and 25.2 ± 0.2° 2-theta measured by CuKa radiation.

4. A process for the preparation of the Crystal modification 5 characterized in claim 3, wherein ruxolitinib hydrochloric acid salt is suspended in a polar protic solvent.

5. The process in accordance with claim 4, characterized in that the polar protic solvent is selected from the group consisting of methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or an organic solvent-water mixture thereof.

6. The process in accordance with claim 5, characterized in that the polar protic solvent is methanol-water mixture, preferably methanol-water mixture ranging from 0 to 40% v/v of methanol in water at room temperature.

7. The process in accordance with claims 4 to 6, characterized in that it comprises the following steps: a/ suspending ruxolitinib hydrochloric acid salt in a a polar protic solvent, preferably the polar protic solvent or an organic solvent-water mixture thereof, selected from the group consisting of: methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or a mixture thereof, more preferably in methanol-water mixture, even more preferably methanol-water mixtures ranging from 0 to 40% v/v of methanol in water at room temperature;

b/ stirring the suspension of step a/ at room temperature for 72 hours;

c/ isolating the Crystal modification 5 of ruxolitinib hydrochloric acid salt.

8. The process in accordance with claim 7, characterized in that it optionally comprises drying of the product of step c/ under the laboratory conditions until the constant weight of the product is reached.

9. A Crystal modification 1 of the crystalline salt in accordance with claims 1 and 2, wherein the acid component (HX) is a phosphoric acid, showing in an X-ray powder diffraction (XRPD) pattern characteristic peaks at about 3.9; 14.5; 15.9; 20.2 and 24.9 ± 0.2° 2-theta measured by CuKct radiation.

10. A process for the preparation of the Crystal modification 1 in accordance with claim 9, characterized in that ruxolitinib free base is dissolved in a polar aprotic solvent and phosphoric acid is then added.

11. The process in accordance with claim 10, characterized in that the polar aprotic solvent is selected from the group comprising methyl-acetate, ethyl-acetate, butyl-acetate, acetonitrile, acetone, methyl ethyl ketone or a mixture thereof.

12. The process in accordance with claim 11, characterized in that the polar aprotic solvent is ethyl-acetate at the temperature of 50°C.

13. The process in accordance with claims 10 to 12, comprising the following steps: a/ dissolution of ruxolitinib free base in a polar aprotic solvent, wherein the polar aprotic solvent is selected from the group consisting of: methyl-acetate , ethyl-acetate, butyl-acetate, acetonitrile, acetone, methyl ethyl ketone or a mixture thereof, preferably the polar aprotic solvent is ethyl-acetate, more preferably it is ethyl-acetate at the temperature of 50°C;

b/ drop-wise addition of the phosphoric acid 85% aqueous solution while precipitation occurred; c/ stirring the suspension of step b/ at 50°C for additional 1 hour;

d/ cooling the suspension of step c/ to a temperature of 0-5°C;

e/ keeping the suspension of step d/ for 16 hours at a temperature of 0-5°C ;

f/ isolating the ruxolitinib phosphoric acid salt in Crystal modification 1.

14. The process in accordance with claim 13, characterized in that it optionally comprises the step of drying the product of step f/ under the laboratory conditions until the constant weight of the product is reached.

15. A Crystal modification 1 of the crystalline salt in accordance with claims 1 and 2, wherein the acid component (HX) is a fumaric acid, showing in an X-ray powder diffraction (XRPD) pattern characteristic peaks at about 5.6; 13.1; 18.1; 22.6; 23.5 and 25.5 + 0.2° 2-theta measured by CuKa radiation.

16. A process for the preparation of the Crystal modification 1 in accordance with claim 15, characterized in that ruxolitinib free base is dissolved in a polar protic solvent and fumaric acid is then added.

17. The process in accordance with claim 16, characterized in that the polar protic solvent is selected from the group consisting of methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or a mixture thereof.

18. The process in accordance with claim 17, characterized in that the polar protic solvent is ethanol, preferably ethanol at the temperature of 50°C.

19. The process in accordance with claims 15 to 17, comprising the following steps: a/ dissolution of ruxolitinib free base in a polar protic solvent preferably selected from the group consisting of: methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or a mixture thereof, preferably in ethanol, more preferably ethanol at the temperature of 50°C; b/ drop-wise addition of the solution of fumaric acid solution in a suitable organic solvent while precipitation occurred;

c/ stirring the suspension of step b/ at 50°C for additional 1 hour;

d/ cooling the suspension of step c/ to a temperature of 0-5°C;

e/ keeping the suspension of step d/ for 16 hours at a temperature of 0-5°C ;

f/ isolating the ruxolitinib fumaric acid salt in Crystal modification 1.

20. The process in accordance with claim 19, characterized in that it optionally comprises the step of drying the product of step f/ under the laboratory conditions until the constant weight of the product is reached.

21. A Crystal modification 1 of the crystalline salt in accordance with claims 1 and 2, wherein the acid component (HX) is a L-tartaric acid, showing in an X-ray powder diffraction (XRPD) pattern characteristic peaks at about 4.1; 8.2; 16.2; 20.9 and 25.5 + 0.2° 2-theta measured by CuKa radiation.

22. A process for the preparation of the Crystal modification 1 in accordance with claim 21, characterized in that ruxolitinib free base is dissolved in a polar protic solvent and L-tartaric acid is then added.

23. The process in accordance with claim 22, characterized in that the polar protic solvent is selected from the group consisting of methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or a mixture thereof.

24. The process in accordance with claim 23, characterized in that the polar protic solvent is ethanol, preferably it is ethanol at the temperature of 50°C.

25. The process in accordance with claims 22 to 24, comprising the following steps: a/ dissolution of ruxolitinib free base preferably in a polar protic solvent that is selected from the group consisting of: methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol or a mixture thereof, more preferably the suitable organic solvent is ethanol, even more preferably it is ethanol at the temperature of 50°C;

b/ drop-wise addition of the solution of L-tartaric acid solution in a suitable organic solvent; c/ stirring the solution of step b/ at 50°C for additional 1 hour;

d/ cooling the solution of step c/ to room temperature;

e/ keeping the solution of step d/ for 16 hours at room temperature while precipitation occurred;

f/ isolating the ruxolitinib L-tartaric acid salt in Crystal modification 1.

26. The process in accordance with claim 25, characterized in that it optionally comprises the step of drying the product of step f/ under the laboratory conditions until the constant weight of the product is reached.

27. A Crystal modification 2 of the crystalline salt in accordance with claims 1 and 2, wherein the acid component (HX) is a L-tartaric acid, showing in an X-ray powder diffraction pattern characteristic peaks at about 6.3; 9.1; 13.3; 16.3; 18.7; 19.4 and 22.8 ± 0.2° 2-theta measured by CuKa radiation.

28. A process for the preparation of the Crystal modification 2 in accordance with claim 27, characterized in that ruxolitinib free base is dissolved in a polar aprotic solvent and L-tartaric acid is then added.

29. The process in accordance with claim 28, characterized in that the polar aprotic solvent is selected from the group consisting of acetone, acetonitrile, methyl ethyl ketone, ethyl acetate, butyl acetate, tetrahydrofuran or a mixture thereof.

30. The process in accordance with claim 29, characterized in that the polar aprotic solvent is acetonitrile, preferably it is acetonitrile at the temperature of 50°C.

31. The process in accordance with claims 28 to 30, comprising the following steps: a/ suspending of ruxolitinib free base in a polar aprotic solvent, preferably the polar aprotic solvent is selected from the group consisting of: acetone, acetonitrile, methyl ethyl ketone; ethyl acetate, butyl acetate, tetrahydrofuran or a mixture thereof, preferably the suitable organic solvent is acetonitrile, more preferably it is acetonitrile at the temperature of 50°C; b/ drop-wise addition of the solution of L-tartaric acid solution in a suitable organic solvent; c/ stirring the solution of step b/ at 50°C for additional 1 hour while precipitation occurred; d/ cooling the suspension of step c/ to room temperature;

e/ keeping the suspension of step d/ for 16 hours at room temperature;

f/ isolating the ruxolitinib L-tartaric acid salt in Crystal modification 2.

32. The process in accordance with claim 31, characterized in that it optionally comprises the step of drying the product of step f/ under the laboratory conditions until the constant weight of the product is reached.

33. A pharmaceutical composition comprising a salt of (3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)pyrazol-l-yl]propanenitrile of Formula I and at least one acid component (HX), characterized in that the acid addition salt is in a crystalline form.

(I)

34. The pharmaceutical composition in accordance with claim 33, characterized in that the acid component (HX) is hydrochloric acid, phosphoric acid, fumaric acid or L-tartaric acid.

35. The pharmaceutical composition in accordance with claims 33 and 34, characterized in that it further comprises one or more pharmaceutically acceptable carriers or excipients.

36. The pharmaceutical composition in accordance with claims 33 to 35, characterized in that it is in a pharmaceutical form suitable for oral administration.

37. The pharmaceutical composition in accordance with claims 33 to 36, characterized in that. the pharmaceutical composition is in a form of a tablet.

38. The crystalline salt in accordance with any of claims 1 to 3, 9, 15, 21 and 27 for use in treating of a disease in a patient wherein said disease is associated with JAK activity.

39. The crystalline salt in accordance with any of claims 1 to 3, 9, 15, 21 and 27 for use in the treatment of cancer.

40. The crystalline salt in accordance with claim 39, characterized in that the cancer is a hematological cancer.

41. The crystalline salt in accordance with any of claims 1 to 3, 9, 15, 21 and 27 for use for treating chronic myelogenous leukemia (C L) in a patient comprising administering to a patient a therapeutically effective amount of the said crystalline salt.

42. The crystalline salt in accordance with any of claims 1 to 3, 9, 15, 21 and 27 for use for treating acute lymphoblastic leukemia (ALL) in a patient comprising administering to a patient therapeutically effective amount of the said crystalline salt.

43. The crystalline salt in accordance with any of claims 1 to 3, 9, 15, 21 and 27 for use for treating chronic myelomonocytic leukemia (CMML) in a patient comprising administering to a patient therapeutically effective amount of the said crystalline salt.