RU2798091C2 - Crystal salts of b-raf-kinase inhibitor - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to crystalline salts of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide, a RAF kinase inhibitor useful in the treatment of cancer and other conditions. The invention relates to a method for obtaining a crystalline monosuccinate salt N-(3-(5-((1-ethylpiperidin-4-yl) (methyl) amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide having a powder X-ray diffraction pattern containing at least five of the following peaks, in units of 2θ: 15.4° ± 0.5°; 16.1° ± 0.5°; 17.2° ± 0.5°; 19.1° ± 0.5°; 19.8° ± 0.5°; 20.0° ± 0.5°; 20.2° ± 0.5°; 20.5° ± 0.5°; 21.5° ± 0.5°; and 21.8° ± 0.5°, including the interaction of N-(3-(5-((1-ethylpiperidin-4-yl) (methyl) amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide with one equivalent of succinic acid in a solvent, where the solvent is selected from C_1-4 alcohol or aqueous C_1-4 alcohol, or from ethyl acetate, where the method includes: heating and stirring N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide with one equivalent of succinic acid in solvent at 30–100°C for 5 min to 4 h with the formation of a suspension, cooling and stirring the suspension to 0–25°C for 5 min to 24 h, filtering the crystalline salt from the suspension, and drying the crystalline salt under reduced pressure at 50°C to give a crystalline salt.

EFFECT: method for obtaining a crystalline monosuccinate salt of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide useful in the treatment of cancer and other diseases associated with RAF kinase activity, which is soluble, stable and non-hygroscopic.

2 cl, 50 dwg, 18 tbl

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No. 62/577,313, filed October 26, 2017, the entire contents of which are incorporated herein by reference.

FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES

The present invention relates to salt forms, and in particular to crystalline salts of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H -pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide (BI 882370), a RAF kinase inhibitor useful in the treatment of cancer and other diseases.

BACKGROUND OF THE INVENTION

Compound N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl )-2,4-difluorophenyl)propane-1-sulfonamide (BI 882370) having the formula I:

I

I

is a RAF kinase inhibitor useful in the treatment of various diseases, including cancer. The compound of formula I, as well as its preparation and use, have been described in patent document WO/2012/104388, which is incorporated herein by reference in its entirety.

The RAS-RAF-MAPK (mitogen-activated protein kinase) signaling pathway plays a critical role in the transmission of proliferation signals generated by transmembrane receptors and cytoplasmic signaling elements to the nucleus. The constitutive activation of this pathway is included in the malignant transformation of some oncogenes. Activating mutations in the RAS family occur in about 15% of cancers, and recent data have shown that B-RAF is mutated in about 7% of cancers (Wellbrock et al., "The RAF proteins take center stage"). ”), Nature Rev. Mol. Cell Biol., 2004, Volume 5, pp. 875-885), identifying it as another important oncogene in this pathway. In mammals, the RAF family of serine/threonine kinases includes three members: A-RAF, B-RAF, and C-RAF. However, activating mutations have so far only been identified in B-RAF, highlighting the importance of this isoform. It appears that B-RAF is the main isoform that binds RAS to MEK, and that C-RAF and A-RAF only signal fine-tuned cellular responses to ERK (Wellbrock et al., Nature Rev. Mol. Cell Biol., 2004, volume 5, pp. 875-885). The most common carcinogenic mutation in B-RAF results in a substitution of valine for glutamic acid at position 600 of the protein (V600E), which significantly increases B-RAF activity, probably because its negative charge mimics a phosphorylation activation loop (Wan et al. , "Mechanism of activation of the RAF-ERK signaling pathway by oncogenic mutations of B-RAF", Cell, 2004, vol. 116, pp. 855-867 ). The largest proportion of B-RAF V600 mutations occurs in malignant melanoma (39%), thyroid cancer (46%), colorectal cancer (10%), bile duct cancer (10%), prostate cancer (4%), ovarian cancer ( 3%), and non-small cell lung cancer (2%), but they also occur at a lower frequency in a variety of other cancers (mutation frequencies according to the COSMIC database (Catalogue Of Somatic Mutations In Cancer; Wellcome Trust Sanger Institute), publication - volume 53, May 15, 2011; http://www.sanger.ac.uk/genetics/CGP/cosmic/). Supported in the literature is the hypothesis that mutated B-RAF ^V600E cancer cells appear to be highly dependent on continuous activation of this pathway—a phenomenon termed "oncogenic addiction"—whereas normal B-RAF ^wt cells use a wider range of signals. This creates an Achilles' heel in that a therapeutic approach can be applied in the treatment of patients with somatically mutated B-RAF ^V600E using oral B-RAF inhibitors.

The key role of B-RAF ^V600E in the distortion of the ERK signaling pathway and thereby tumorigenesis has been demonstrated in several independent experimental approaches, such as overexpression of oncogenically mutated B-RAF in vitro and in vivo (Wan et al., Cell, 2004, vol. 116, p. 855-867; Wellbrock et al., Cancer Res., 2004, vol. 64: pp. 2338-2342), in vitro siRNA knockdown (Karasarides et al., Oncogene, "V599EB-RAF is an oncogene in melanocytes"(" V599EB-RAF as an oncogene in melanocytes"), 2004, vol. 23, pp. 6292-6298), or in inducible short RNA xenograft models, where a dominant-negative B-RAF signaling pathway was found highly associated with in vivo carcinogenicity ( Hoeflich et al., "Oncogenic BRAF is required for tumor growth and maintenance in melanoma models", Cancer Res., 2006, vol. 66, pp. 999-1006) .

Treatment of melanoma with B-RAF ^V600E mutation or colon cancer cells is to promote phenotypic B-RAF inhibition (eg, decrease phospho-MEK and phospho-ER levels, decrease cyclin D expression, and stimulate p27 protein expression). Thus, these cells are blocked in the G1 phase of the cell cycle and do not proliferate.

Clinical evidence of mechanism and proof of concept were obtained in the treatment of cancer in patients with B-RAF mutation ^V600E melanoma using Zelboraf®, a B-RAF inhibitor (PLX-4032, vemurafenib, from Plexxikon/Daiichi Sankyo/Roche. Bollag et al. ., "Clinical efficacy of a RAF inhibitor needs broad target blockade in BRAF-mutant melanoma", Nature, 2010, vol. 467(7315), p. 596-599 Flaherty et al., New Engl. J. Med., "Inhibition of Mutated, Activated BRAF in Metastatic Melanoma", 2010, vol. 363, p. 809- 819; Chapman et al., "Improved Survival with Vemurafenib in Melanoma with BRAF V600E Mutation", New Engl. J. Med., 2011, vol. 364: p. 2507 -2516 Favorable treatment rates were observed in both Phase I and Phase III clinical trials. Melanoma patients with B-RAF ^V600K mutation have also been reported to respond to therapy (Rubinstein et al., "Incidence of the V600K mutation among melanoma patients with BRAF mutations, and potential therapeutic response to the specific BRAF inhibitor PLX4032", ( "Incidence with V600K codon mutation among melanoma patients with BRAF mutations, and potential therapeutic response to a specific BRAF inhibitor PLX4032", J. Transl. Med., 2010, vol. 8, p. 67).

The most common B-RAF mutation is the substitution of valine for glutamate at amino acid residue 600, with more than 90% of all B-RAF mutations (Wellbrock et al., Nature Rev. Mol. Cell Biol., 2004, volume 5, pp. 875-885), the second most common mutation is valine to lysine, other mutations have been found at this position with less frequency (Wellbrock et al., Nature Rev. Mol. Cell Biol., 2004, vol. 5, p. 875 -885, and mutation rates according to the COSMIC database (Catalogue Of Somatic Mutations In Cancer; Wellcome Trust Sanger Institute) publication - volume 53, May 15, 2011; http://www.sanger.ac.uk/genetics/CGP/cosmic/ ). Additional mutations have been found, for example, in the glycine-rich loop (Wellbrock et al., Nature Rev. Mol. Cell Biol., 2004, vol. 5, pp. 875-885). It appears that not all of these rare mutations result in direct activation of B-RAF (Wan et al., "Mechanism of activation of the RAF-ERK signaling pathway by oncogenic mutations of B-RAF"). signaling pathway by oncogenic B-RAF mutations”), Cell, 2004, volume 116, pp. 855-867).

The compound of formula I is a potent and selective RAF inhibitor that binds to the DFG-out (inactive) B-RAF kinase conformation. The compound inhibits the proliferation of human B-RAF-mutated melanoma cells with 100-fold higher activity (1-10 nmol/L) than vemurafenib, while no effect on wild-type cells was seen at 1000 nmol/L. The compound oral solution was efficacious in mouse models of B-RAF-mutated melanoma and colorectal cancer, and at 25 mg/kg twice daily showed superior efficacy compared to vemurafenib, dabrafenib, or trametinib. The compound was also active in A375 melanoma mice that were resistant to vemurafenib, particularly when administered in combination with trametinib. The compound treated mouse did not show any weight loss or clinical symptoms of intolerance, and no pathological changes were observed in several major organs examined, including the skin. In addition, in an experimental study in rats (up to 60 mg/kg daily for 2 weeks), the compound showed no toxicity in terms of clinical biochemistry, hematology, pathology and toxicogenomics. These results are described in Waizenegger et al., Mol. Cancer Ther., 2016, Volume 15 (No. 3); pp. 354-365, the entire content of which is incorporated into this description by reference.

It may be advantageous to use a form of the drug having excellent stability or other desirable formulation properties, such as one or more salts or crystalline forms of the drug, for preparation, purification and formulation of the drug. The formation of salts of basic or acidic drugs can sometimes lead to drug forms that have favorable properties such as solubility, non-hygroscopicity, crystallinity, and other physical properties that are favorable for drug formulation. On the other hand, identifying a suitable salt or other crystalline form that is suitable for drug formulation is difficult because there are many variables in terms of salt or crystalline form formation. These include the existence of a variety of possible acids and bases that could be used as a counterion, a variety of stoichiometric ratios that might be possible for a given basic or acidic drug to be coupled with an acidic or basic counterion, a wide variety of solvents and solvent systems (including including solvent combinations) that can potentially be used in attempts to form salts or crystal forms, and a variety of conditions (such as temperature, or heating or cooling conditions) under which salts or crystal forms can be formed. All of these variable factors can affect the properties of the salts or crystal forms that might be obtained. Salts or solid forms may also have a variety of properties that make them unsuitable for drug development and formulation, such as the absence of crystallinity (amorphous forms), the presence or formation of multiple crystalline forms, which may be interconvertible and/or have different properties ( polymorphism), lack of solubility in water, hygroscopicity or stickiness of the solid. In addition, the formation of salts and crystalline forms and their properties are generally highly unpredictable.

Accordingly, the crystalline salt forms of the compound of formula I provided herein help to meet the continuing need for the development of a RAF kinase inhibitor for the treatment of serious diseases.

SUMMARY OF THE INVENTION

The present invention provides N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monosuccinate salt -1-yl)-2,4-difluorophenyl)propan-1-sulfonamide and specific crystalline forms of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-( pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

In some embodiments, the crystalline salt is substantially anhydrous.

In some embodiments, the crystalline salt is substantially unsolvated.

In some embodiments, the crystalline salt is N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[ 3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

In some embodiments, the crystalline salt has a powder X-ray diffraction pattern containing at least one of the following peaks, in units of 2θ: 15.4±0.5°; 20.0±0.5°; and 21.8±0.5°. In some embodiments, peak at 15.4 ±0.5°; 20.0±0.5°; 21.8±0.5° is the peak with the highest relative intensity in the X-ray powder diffraction pattern.

In some embodiments, the crystalline salt has an x-ray powder diffraction pattern containing at least one of the following peaks, in 2θ units: 15.4±0.2°; 20.0±0.2°; and 21.8±0.2°. In some embodiments, peak at 15.4 ±0.2°; 20.0±0.2°; or 21.8±0.2° is the peak with the highest relative intensity in the X-ray powder diffraction pattern.

In some embodiments, the crystalline salt has a powder X-ray diffraction pattern containing at least one peak, in units of 2θ, at 15.4±0.5°. In some embodiments, the peak at 15.4±0.5° is the peak with the highest relative intensity in the powder x-ray diffraction pattern.

In some embodiments, the crystalline salt has a powder X-ray diffraction pattern containing at least one peak, in units of 2θ, at 15.4±0.2°. In some embodiments, the peak at 15.4±0.2° is the peak with the highest relative intensity in the powder x-ray diffraction pattern.

In some embodiments, the crystalline salt has a powder X-ray diffraction pattern containing the following peaks, in 2θ units: 15.4±0.5°; 20.0±0.5°; and 21.8±0.5°. In some embodiments, peak at 15.4 ±0.5°; 20.0±0.5°; 21.8±0.5° is the peak with the highest relative intensity in the X-ray powder diffraction pattern.

In some embodiments, the crystalline salt has a powder X-ray diffraction pattern containing the following peaks, in 2θ units: 15.4±0.2°; 20.0±0.2°; and 21.8±0.2°. In some embodiments, peak at 15.4 ±0.2°; 20.0±0.2°; or 21.8±0.2° is the peak with the highest relative intensity in the X-ray powder diffraction pattern.

In some embodiments, the crystalline salt has an x-ray powder diffraction pattern substantially as shown in FIG. 10 FIG. 26 or FIG. 38.

In some embodiments, the crystalline salt has a differential scanning calorimetry (DSC) thermogram substantially as shown in FIG. 11 FIG. 27 or FIG. 39.

In some embodiments, the crystalline salt has thermogravimetric analysis (TGA) results substantially as shown in FIG. 12 FIG. 28 or FIG. 39.

In some embodiments, the crystalline salt is substantially isolated.

The present invention provides a composition (e.g. a pharmaceutical composition) comprising N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H crystalline monosuccinate salt -pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide, or any embodiments thereof. The composition may include at least one pharmaceutically acceptable carrier.

The present invention provides a dosage form containing crystalline N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3, 2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide, or any embodiments thereof, or a composition containing a crystalline salt, or any embodiments thereof. The dosage form may be in the form of a tablet.

The present invention provides a process for the preparation of crystalline N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2- b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide. The method includes the interaction of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1 -yl)-2,4-difluorophenyl)propane-1-sulfonamide with one equivalent of succinic acid. The method may include crystallizing or recrystallizing the salt from a C _1-4 alcohol, an aqueous C _1-4 alcohol, or ethyl acetate. The method may include crystallizing or recrystallizing the salt from ethanol, isopropanol, aqueous ethanol or aqueous isopropanol, or from ethyl acetate.

The present invention provides a method of treating a disease in a patient in which the disease is associated with abnormal RAF kinase expression or activity, comprising administering to the patient a therapeutically effective amount of N-(3-(5-((1-ethylpiperidin-4-yl)( methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide, or any embodiments thereof, or a composition comprising N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b ]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide. The present invention also provides N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b ]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide, or any embodiment thereof, for use in the treatment of a disease associated with abnormal RAF kinase expression or activity. Also presented is the use of crystalline monosuccinate salt N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide, or any embodiment thereof, for the treatment of a disease associated with abnormal RAF kinase expression or activity. Also presented is the use of crystalline monosuccinate salt N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide, or any embodiments thereof, for the preparation of a medicament for the treatment of a disease associated with abnormal RAF kinase expression or activity.

In some embodiments, the disease is associated with abnormal expression or activity of B-RAF kinase. In some embodiments, the B-RAF kinase is a mutated B-RAF kinase. In some embodiments, the B-RAF kinase is a V600E mutated B-RAF kinase. In some embodiments, the disease is selected from cancer, infectious diseases, inflammation, and autoimmune diseases.

In addition, the present invention provides a method of treating cancer comprising administering to a patient a therapeutically effective amount of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5- yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide, or any embodiment thereof, or a composition comprising crystalline N-(3- (5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4- difluorophenyl)propane-1-sulfonamide. The present invention also provides N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b ]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide, or any embodiment thereof, for use in the treatment of cancer. Also provided is the use of crystalline N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide, or any embodiment thereof, for the treatment of cancer. Also provided is the use of crystalline N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide, or any embodiments thereof, to obtain a medicament for the treatment of cancer.

In some embodiments, the cancer is breast, prostate, colon, endometrial, brain, bladder, skin, uterine, ovarian, lung, pancreatic, kidney, stomach, or blood cancer. In some embodiments, the cancer is malignant melanoma, thyroid cancer, colorectal cancer, bile duct cancer, prostate cancer, ovarian cancer, or non-small cell lung cancer.

In some embodiments, cancer is associated with RAF kinase expression or activity. In some embodiments, the cancer is associated with the expression or activity of B-RAF kinase. In some embodiments, the B-RAF kinase is a mutated B-RAF kinase. In some embodiments, the B-RAF kinase is a V600E mutated B-RAF kinase. In some embodiments, the cancer is melanoma. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is thyroid cancer. In some embodiments, the cancer expresses a mutated B-RAF kinase. In some embodiments, the cancer expresses a V600E mutated B-RAF kinase.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an X-ray powder diffraction pattern (XRPD) of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3, 2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 2A is an image of a multiwell plate showing N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2- b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide obtained with various acids and solvents and isolated by crystallization in a suspension experiment.

FIG. 2B is a series of images of XRPD scans taken from each of the wells of the multiwell plate of FIG. 2A showing the crystallinity of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine salts -1-yl)-2,4-difluorophenyl) propane-1-sulfonamide obtained by forming a suspension with various acids and solvents.

FIG. 3A is an image of an evaporation plate showing N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2- b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide prepared with various acids and solvents and isolated by crystallization in an evaporation experiment.

FIG. 3B is a series of images of XRPD scans taken from each of the wells of the multiwell plate of FIG. 3A showing the crystallinity of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine salts -1-yl)-2,4-difluorophenyl) propane-1-sulfonamide obtained by evaporation with various acids and solvents.

FIG. 4 is a series of images of XRPD scans performed on samples of various crystalline salts of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H- pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 5 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide with the sample from well E2 (lower graph) compared to the sample obtained from scaled synthesis (upper graph).

FIG. 6 is a DSC plot of the monohydrochloride salt of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide.

FIG. 7 is a TGA plot of the monohydrochloride salt of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide.

FIG. 8 is a sorption-desorption plot for N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide showing weight gain and loss when the relative humidity ranged from 0-100%.

FIG. 9 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide before (lower graph) and after (upper graph) the sorption-desorption experiment.

FIG. 10 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide with the sample from well F10 (lower graph) compared to the sample obtained from scaled synthesis (upper graph).

FIG. 11 is a DSC plot of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide.

FIG. 12 is a TGA plot of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide.

FIG. 13 is a sorption-desorption plot for N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide showing weight gain and loss when the relative humidity ranged from 0-100%.

FIG. 14 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide before (lower graph) and after (upper graph) the sorption-desorption experiment, showing that no significant change in the structure occurred.

FIG. 15 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide.

FIG. 16 is a DSC plot of the monohydrochloride salt of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide. The sample showed an endotherm with the beginning of the change at about 313°C. As a rule, the samples had an endotherm with the beginning of the change in the range of 312-322°C, corresponding to the melting point. Therefore, the melting point was measured at a value of about 317±5°C. Some samples had an additional endothermic event at about 250° C. corresponding to the loss of the solvent.

FIG. 17 is a TGA plot of the monohydrochloride salt of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide. The sample exhibited a loss on drying of about 1.7-2.5% up to about 250°C, corresponding to the release of water and solvent. The water content was measured at about 0.9% (Karl Fischer method).

FIG. 18 is a plot of the sorption-desorption isotherm for N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b ]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide showing weight gain and loss when the relative humidity ranged from 0-90%.

FIG. 19 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide before (lower graph) and after (upper graph) the sorption-desorption experiment, showing that no significant change in the structure occurred.

FIG. 20 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide after grinding in a shaking mill for 10 minutes at 30 rpm.

FIG. 21 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide after pressing at 2000 N for 1 second to form a tablet (5 mm diameter).

FIG. 22 represents ¹ H-NMR spectrum (400 MHz, DMSO-d6) of the monohydrochloride salt of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl )-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 23 is a fragment of the ¹ H-NMR spectrum (400 MHz, DMSO-d6) in the aliphatic region (δ 0.5-6.0) of the monohydrochloride salt of N-(3-(5-((1-ethylpiperidin-4-yl) (methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 24 is a fragment of the ¹ H-NMR spectrum (400 MHz, DMSO-d6) in the aromatic region (δ 6.0-11.5) of the monohydrochloride salt of N-(3-(5-((1-ethylpiperidin-4-yl) (methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 25 is a graph showing the dissolution rate of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2] monohydrochloride salt -b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide over a period of 0-60 minutes in an aqueous medium at pH values from 1.0 to 7.4.

FIG. 26 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide.

FIG. 27 is a DSC plot of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide.

FIG. 28 is a TGA plot of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide.

FIG. 29 is a graph of the sorption-desorption isotherm for N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b ]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide showing weight gain and loss when the relative humidity ranged from 0-90%.

FIG. 30 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide before (lower graph) and after (upper graph) the sorption-desorption experiment, showing that no significant change in the structure occurred.

FIG. 31 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide after grinding in a shaking mill for 10 minutes at 30 rpm.

FIG. 32 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide after pressing at 2000 N for 1 second to form a tablet (5 mm diameter).

FIG. 33 represents ¹ H-NMR spectrum (400 MHz, DMSO-d6) of the monosuccinate salt of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl )-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 34 is a fragment of the ¹ H-NMR spectrum (400 MHz, DMSO-d6) in the aliphatic region (δ 0.5-6.0) of the monosuccinate salt of N-(3-(5-((1-ethylpiperidin-4-yl) (methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 35 is a fragment of the ¹ H-NMR spectrum (400 MHz, DMSO-d6) in the aromatic region (δ 6.0-10.0) of the monosuccinate salt of N-(3-(5-((1-ethylpiperidin-4-yl) (methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 36 is a graph showing the dissolution rate of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2] monohydrochloride salt -b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide for a period of 0-60 minutes in an aqueous medium at pH values from 1.0 to 7.4.

FIG. 37 is a plot of plasma concentration versus time values after oral administration to rats of (A) N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl) monohydrochloride salt )-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide at 40 mg/kg; (B) N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monohydrochloride salt -1-yl)-2,4-difluorophenyl) propane-1-sulfonamide at 80 mg/kg; (C) N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monosuccinate salt -1-yl)-2,4-difluorophenyl) propane-1-sulfonamide at 40 mg/kg; (D) N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monosuccinate salt -1-yl)-2,4-difluorophenyl) propane-1-sulfonamide at 80 mg/kg.

FIG. 38 is an XRPD plot of two batches of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3, monosuccinate salt Form A, 2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 39 is a pair of plots of TGA and DSC analysis of Form A of the monosuccinate salt N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H- pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 40 represents ¹ H-NMR spectrum of (DMSO-d6) N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl) monosuccinate salt Form A -1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 41 represents ¹ H-NMR spectrum (deuterated methanol) of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-, monosuccinate salt Form A 1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 42 is a pair of XRPD plots of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2] monosuccinate salt Form A -b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide before and after heating to 170° C. and cooling to room temperature.

FIG. 43 is a pair of TGA plots of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2] monosuccinate salt Form A -b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide before and after heating to 170° C. and cooling to room temperature.

FIG. 44 is a pair of DSC plots of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2] monosuccinate salt Form A -b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide before and after heating to 170° C. and cooling to room temperature.

FIG. 45 represents ¹ H-NMR spectrum of (DMSO-d6) N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl) monosuccinate salt Form A -1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide before and after heating to 170° C. and cooling to room temperature.

FIG. 46 is a series of XRPD plots showing that Form A of the N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H- pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide does not change during the equilibrium dissolution test.

FIG. 47 is a series of XRPD plots showing the stability of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[] monosuccinate salt Form A 3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide after storage under various conditions.

FIG. 48 is a series of DVS isotherm plots showing the low hygroscopicity of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H monosuccinate salt Form A -pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 49 is a pair of XRPD plots showing that N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo monosuccinate salt Form A [3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide does not change shape after DVS.

FIG. 50 is a pair of polarized light microscopy images of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo monosuccinate salt Form A [3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

DETAILED DESCRIPTION OF THE INVENTION

I. Definitions

Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the field of technology to which this invention pertains.

For the terms "for example" and "such as" and their grammatical equivalents, the expression "and without limitation" is understood accordingly, unless expressly stated otherwise.

As used here, the term "about" means "approximately" (eg, plus or minus about 10% of the specified value).

At various points in this specification, certain features may be disclosed in groups or ranges. Such a representation is specifically intended to include each and every individual subcombination of members of such groups and ranges. For example, the term "C _1-4 -alkyl" specifically means the individual disclosure of (without limitation) methyl, ethyl, C ₃ -alkyl and C ₄ -alkyl groups.

The terms "individual", "subject", or "patient", used interchangeably, refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, and most preferably humans.

The term "treatment" or "therapy" means one or more of (1) prevention of a disease; for example, preventing a disease, condition, or disorder in an individual who may be predisposed to the disease, condition, or disorder but is not yet experiencing or showing signs of pathology or symptoms of the disease; (2) disease suppression; for example, suppression of a disease, condition, or disorder in an individual who is experiencing or exhibiting signs of pathology or symptoms of the disease, condition, or disorder (ie, arresting or slowing the further development of the pathology and/or symptomatology); and (3) reducing the intensity of the disease; for example, improvement in a disease, condition, or disorder in an individual who is experiencing or exhibiting signs of pathology or symptoms of the disease, condition, or disorder (i.e., reversal of the pathology and/or symptomatology), such as a reduction in the severity of the disease.

The expression "therapeutically effective amount" means the amount of an active salt or crystalline form of a pharmaceutical preparation that elicits a biological or medical response that is elicited in a tissue, system, animal, individual, or human by a researcher, veterinarian, medical professional, or other medical practitioner.

Certain features of the invention, which, for the sake of clarity, are described in the context of separate embodiments, may also be presented in combination in a single embodiment. On the contrary, various features of the invention, which, for brevity, are described in the context of a single embodiment, may also be presented individually or in any suitable subcombination.

Abbreviations

The following abbreviations and symbols may be used in the present invention: Ac (acetyl); aq. (water); Boc (tert-butoxycarbonyl); Bu (butyl); °C (degrees Celsius); c (concentration); conc. (concentrated); d (day(s)); DCM (dichloromethane); DEA (diethylamine); DIPEA (N-ethyl-N, N-diisopropylamine (Hünig's base)); DMF (N,N-dimethylformamide); DMSO (dimethyl sulfoxide); DSC (differential scanning calorimetry); DVS (dynamic vapor sorption); EDTA (ethylenediaminetetraacetic acid); EGTA (ethylene glycol tetraacetic acid); eq. (equivalent(s)); ESI (spray ionization in an electric field, electrospray); Et (ethyl); Et ₂ O (diethyl ether); EtOAc (ethyl acetate); EtOH (ethanol); g (gram(s)); h (hour(s)); HPLC (high performance liquid chromatography); HCl (hydrochloric acid); i (iso-); iPrOH (isopropanol); L (litre(s)); LC (liquid chromatography); M (molar) ; mg (milligram(s)) Me (methyl); MeCN (acetonitrile); MeOH (methanol); min. (minutes); mL (milliliter); mM (millimolar); MPLC (medium pressure liquid chromatography); MS ( mass spectrometry); NP (normal phase); Ph (phenyl); Pr (propyl); Py (pyridine); rac (racemic); Rf (retention factor); RH (relative humidity); RP (opposite phase); rps ( revolutions per second); rt (ambient temperature); tBu (tert-butyl); TEA (triethylamine); temp. (temperature); tert (tertiary); Tf (triflate); TFA (trifluoroacetic acid); TGA (thermogravimetric analysis ); THF (tetrahydrofuran); TLC (thin layer chromatography); tRet. (retention time (HPLC)); UV (ultraviolet), XRPD (powder x-ray diffractometry) Other conventional abbreviations may also apply here.

II. Crystal salts

The present invention relates, inter alia, to salt forms, and in particular to crystalline salt forms of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5 -yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide (BI 882370) having formula I:

I

I

which are useful, for example, in the preparation of solid dosage forms of the above compound for the treatment of various diseases, including cancer.

Different salt and crystalline forms of the same substance may have different bulk characteristics related to, for example, hygroscopicity, solubility, stability, and the like. Forms with high melting points may have good thermodynamic stability, which may be beneficial in extending the shelf life of drug formulations containing a particular salt or crystalline form. Forms with lower melting points may be less thermodynamically stable, but are advantageous in that they have increased aqueous solubility, which results in increased drug bioavailability. Forms that have low hygroscopicity may be desirable due to their stability under conditions of heat and humidity and resistance to degradation during long-term storage. Anhydrous forms may be desirable because they can be uniformly prepared without problems with variation in weight or composition due to variation in solvent or water content. On the other hand, hydrated or solvated forms may sometimes be preferred if they are less hygroscopic and exhibit improved moisture stability under storage conditions.

As used herein, "crystal form" is intended to mean a specific configuration of the crystal lattice of a crystalline substance (which may include the salts described herein). Different crystalline forms of the same substance may have different crystal lattices (for example, unit cells), which have different physical properties that are characteristic of each of the crystalline forms. In some situations, different lattice configurations have different water or solvent contents. Different crystal lattices can be identified by solid state characterization methods such as X-ray powder diffraction (XRPD). Other characterization methods such as differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), dynamic vapor sorption (DVS); Solid-state NMR spectroscopy, and the like, further aid in the identification of crystal form as well as assist in determining stability and solvent/water content.

Crystalline forms of the material may include both solvated (eg, hydrated) and non-solvated (eg, anhydrous) forms. The hydrated form is a crystalline form that includes water in the crystal lattice. Hydrated forms may be stoichiometric hydrates where water is present in the lattice at a specific water-molecular ratio, such as for hemihydrates, monohydrates, dihydrates, etc. Hydrated forms can also be non-stoichiometric, where the water content is variable and dependent on external conditions such as humidity.

Crystalline forms can be characterized by X-ray powder diffraction (XRPD). The pattern of XRPD reflections (peaks, diffraction maxima) is usually considered as a characteristic feature of the structure of a particular crystalline form. It is well known that the relative intensities of the XRPD peaks can vary widely depending on, among other things, the sample preparation method, crystal fineness, filters, sample mounting technique, and the particular instrument used. In some cases, new peaks may be observed, or existing peaks may disappear, depending on the instrument type or settings (eg, whether a Ni filter is applied or not). As used herein, the term "peak" refers to a reflex having a relative height/intensity of at least about 4% of the height/intensity of the maximum peak. Moreover, instrument variation and other factors can affect 2-theta values. Thus, peak assignments such as those described herein may vary by plus or minus about 0.2° (2-theta), and the term "substantially" as used herein in the context of XRPD is intended to include the above variations.

Similarly, temperature readings in connection with differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), or other thermal experiments may vary by about ±4°C depending on the instrument, specific settings, sample acquisition conditions, etc. d. For example, with respect to DSC, it is known that the observed temperatures will depend on the rate of temperature change, as well as the sample preparation method and the specific instrument used. Thus, the values described here in relation to DSC thermograms may vary, as indicated above, by ±4°C. Accordingly, a crystal form described herein having a DSC thermogram "essentially" as shown in any of the Figures is understood to include such variation.

The salts described herein may be isolated in various crystalline forms, which may include crystalline forms that are anhydrous, hydrated, unsolvated, or solvated. Exemplary hydrates include hemihydrates, monohydrates, dihydrates, and the like. In some embodiments, the crystalline forms are anhydrous and unsolvated. By "anhydrous" is meant that the crystalline form of the compound of formula I essentially does not contain bound water in the crystal lattice structure, that is, the compound does not form a crystalline hydrate.

In some embodiments, the salts and crystalline forms of the invention may be substantially isolated. By "substantially isolated" is meant that the particular salt or crystalline form of the compound is at least partially free of impurities. For example, in some embodiments, the salt or crystalline form of the invention comprises less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 15%, less less than about 10%, less than about 5%, less than about 2.5%, less than about 1%, or less than about 0.5% impurities. Typically, impurities include any contaminant that is not essentially isolated salt or crystalline form, including, for example, other salts or other crystalline forms, and other substances.

In some embodiments, the salt or crystalline form is substantially free of other crystalline forms. The expression "essentially does not contain other crystalline forms" means that a particular crystalline form includes more than about 80%, more than about 90%, more than about 95%, more than about 98%, more than about 99% , or more than about 99.5% by weight of a particular crystalline form.

The present invention provides crystalline N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide, and specific crystalline forms of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3 monosuccinate salt -(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

The term "monosuccinate salt" means that the components of succinic acid and base N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo [3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide present in the salt are in an acid:base ratio of about 1:1, for example, with a ratio ranging from about 0.8:1 to about 1.2:1, about 0.9:1 to about 1.1:1, about 1:1.2 to about 1:0.8, or about 1:1 .1 to about 1:0.9, for example, in a ratio of about 0.8:1, about 0.9:1, about 1:1, or about 1.1:1, or about 1:0.8, about 1:0.9, about 1:1, about 1:1.1 or about 1:1.2.

As detailed further below, N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2 -b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide has unanticipated properties such as improved solubility, improved intrinsic dissolution rate, and improved pharmacokinetic characteristics compared to the free base and other N-salt forms. (3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2 ,4-difluorophenyl) propane-1-sulfonamide.

In some embodiments, the crystalline salt is substantially anhydrous. By "substantially anhydrous" is meant that the crystalline salt contains less than a stoichiometrically equivalent amount of water and does not contain water as part of the salt's crystalline structure. In some embodiments, water, if present, is present in an amount of about 5% or less, about 4% or less, about 3% or less, about 2% or less, about 1% or less, about 0.5% or less, about 0.2% or less, or about 0.1% or less, by weight of the crystalline salt. The use of the term "substantially anhydrous" does not exclude the presence of trace amounts of water.

In some embodiments, the crystalline salt is substantially unsolvated. By "substantially unsolvated" is meant that the crystalline salt contains less than a stoichiometrically equivalent amount of solvent and does not contain solvent molecules as part of the salt's crystalline structure. In some embodiments, the solvent, if present, is present in an amount of about 5% or less, about 4% or less, about 3% or less, about 2% or less, about 1% or less, about 0.5% or less, about 0.2% or less, or about 0.1% or less, by weight of the crystalline salt. The use of the term "essentially unsolvated" does not exclude the presence of trace amounts of solvent.

In some embodiments, the crystalline salt is substantially free of solvents other than water. By "substantially free" is meant that the crystalline salt contains less than a stoichiometrically equivalent amount of a solvent other than water and does not contain a molecule of a solvent other than water as part of the crystalline structure of the salt. In some embodiments, a solvent other than water, if present, is present in an amount of about 5% or less, about 4% or less, about 3% or less, about 2% or less, about 1% or less, about 0.5 % or less, about 0.2% or less, or about 0.1% or less by weight of the crystalline salt. The use of the term "substantially free of solvents other than water" does not preclude the presence of trace amounts of such solvents.

In some embodiments, the crystalline salt is N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[ 3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide (as further described in more detail below).

The crystalline form of the salt is identified by unique characteristics with respect to, for example, X-ray powder diffractometry (XRPD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and dynamic vapor sorption (DVS).

In some embodiments, N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2- b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide is characterized by an XRPD pattern essentially as shown in FIG. 10 FIG. 26 or FIG. 38.

In some embodiments, N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2- b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide was characterized by an XRPD pattern essentially as shown in Table 9.

In some embodiments, the crystalline salt has a powder X-ray diffraction pattern containing at least one of the following peaks, in units of 2θ: 15.4±0.5°; 20.0±0.5°; and 21.8±0.5°. In some embodiments, peak at 15.4 ±0.5°; 20.0±0.5°; 21.8±0.5° is the peak with the highest relative intensity in the X-ray powder diffraction pattern.

In some embodiments, the crystalline salt has an x-ray powder diffraction pattern containing at least one of the following peaks, in 2θ units: 15.4±0.2°; 20.0±0.2°; and 21.8±0.2°. In some embodiments, peak at 15.4 ±0.2°; 20.0±0.2°; or 21.8±0.2°, represents the peak with the highest relative intensity in the X-ray powder diffraction pattern.

In some embodiments, the crystalline salt has a powder X-ray diffraction pattern containing at least one peak, in units of 2θ, at 15.4±0.5°. In some embodiments, the peak at 15.4±0.5° is the peak with the highest relative intensity in the powder x-ray diffraction pattern.

In some embodiments, the crystalline salt has a powder X-ray diffraction pattern containing at least one peak, in units of 2θ, at 15.4±0.2°. In some embodiments, the peak at 15.4±0.2° is the peak with the highest relative intensity in the powder x-ray diffraction pattern.

In some embodiments, the crystalline salt has a powder X-ray diffraction pattern containing the following peaks, in 2θ units: 15.4±0.5°; 20.0±0.5°; and 21.8±0.5°. In some embodiments, peak at 15.4 ±0.5°; 20.0±0.5°; or 21.8±0.5°, represents the peak with the highest relative intensity in the powder x-ray diffraction pattern.

In some embodiments, the crystalline salt has a powder X-ray diffraction pattern containing the following peaks, in 2θ units: 15.4±0.2°; 20.0±0.2°; and 21.8±0.2°. In some embodiments, peak at 15.4 ±0.2°; 20.0±0.2°; or 21.8±0.2° is the peak with the highest relative intensity in the X-ray powder diffraction pattern.

In some embodiments, the crystalline salt has an X-ray powder diffraction pattern containing at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all of next peaks, 2θ units: 15.4 ±0.5°; 16.1±0.5°; 17.2±0.5°; 19.1±0.5°; 19.8±0.5°; 20.0±0.5°; 20.2±0.5°; 20.5±0.5°; 21.5±0.5°; and 21.8±0.5°. In some embodiments, the powder x-ray diffraction pattern contains a peak of 15.4±0.5°; 20.0±0.5°; or 21.8 ± 0.5° as the peak with the highest relative intensity in the X-ray powder diffraction pattern.

In some embodiments, the powder x-ray diffraction pattern contains a peak at 15.4±0.5° as the peak with the highest relative intensity in the powder x-ray diffraction pattern.

In some embodiments, the crystalline salt has an X-ray powder diffraction pattern containing at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all of next peaks, 2θ units: 15.4 ±0.2°; 16.1±0.2°; 17.2±0.25°; 19.1±0.2°; 19.8±0.2°; 20.0±0.2°; 20.2±0.2°; 20.5±0.2°; 21.5±0.2°; and 21.8±0.2°. In some embodiments, the powder x-ray diffraction pattern contains a peak of 15.4±0.2°; 20.0±0.2°; or 21.8 ± 0.2° as the peak with the highest relative intensity in the X-ray powder diffraction pattern. In some embodiments, the powder x-ray diffraction pattern contains a peak at 15.4±0.2° as the peak with the highest relative intensity in the powder x-ray diffraction pattern.

In some embodiments, the powder x-ray diffraction pattern contains at least two of the peaks at 15.4±0.5°; 20.0±0.5°; or 21.8±0.5°, as the peaks with the highest relative intensity in the X-ray powder diffraction pattern.

In some embodiments, the powder x-ray diffraction pattern contains at least three of the peaks at 15.4±0.5°; 20.0±0.5°; or 21.8±0.5°, as the peaks with the highest relative intensity in the X-ray powder diffraction pattern.

In some embodiments, the powder x-ray diffraction pattern contains at least two of the peaks at 15.4±0.2°; 20.0±0.2°; or 21.8 ± 0.2°, as the two peaks with the highest relative intensity in the X-ray powder diffraction pattern.

In some embodiments, the powder x-ray diffraction pattern contains at least peaks at 15.4±0.2°; 20.0±0.2°; or 21.8 ± 0.2°, as the three peaks with the highest relative intensity in the X-ray powder diffraction pattern.

In some embodiments, the crystalline salt has a differential scanning calorimetry (DSC) thermogram substantially as shown in FIG. 11 FIG. 27 or FIG. 39.

In some embodiments, the crystalline salt has a thermogravimetric analysis (TGA) result substantially as shown in FIG. 12 FIG. 28 or FIG. 39.

In some embodiments, the crystalline salt is substantially isolated. In some embodiments, the crystalline salt is at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% of Form A monosuccinate salt N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1- yl)-2,4-difluorophenyl)propane-1-sulfonamide

Monosuccinate salt N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1- yl)-2,4-difluorophenyl)propane-1-sulfonamide can be obtained by the reaction of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl )-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide with an appropriate amount, typically about one equivalent, of succinic acid.

The reaction can be carried out in a suitable solvent. The reaction can be carried out by dissolving N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine -1-yl)-2,4-difluorophenyl)propane-1-sulfonamide and adding succinic acid, which can also be dissolved in acid. If necessary, heating of solutions of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2- b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide and/or succinic acid to dissolve the compound. Heating may be carried out to a temperature above room temperature, for example, about 30°C, about 40°C, about 50°C, about 60°C, about 70°C, about 80°C, about 90°C, or about 100 °C. The reaction may be carried out over a period of time, such as about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 40 minutes, about 50 minutes, about 1 hour, about 2 hours, about 3 hours, or about 4 hours. After heating, the solution may be cooled, for example, to a temperature at or below room temperature, for example, about 25°C, about 20°C, about 15°C, about 10°C, about 5°C, or about 0°C. . After such cooling, the reaction mixture may be held for an additional period of time at a lower temperature, for example, for about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 40 minutes, about 50 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 8 hours, about 16 hours, or about 24 hours.

During the steps to form the salt or its crystalline form, the solution or suspension in which the reaction is carried out may be agitated, eg stirred.

Suitable solvents for the formation of the N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine salt -1-yl)-2,4-difluorophenyl)propane-1-sulfonamide with succinic acid, including Form A salts, include C _1-4 alcohol, aqueous C _1-4 alcohol, or ethyl acetate. Solvents may include methanol, ethanol, isopropanol, aqueous methanol, aqueous ethanol, aqueous isopropanol, or ethyl acetate.

Crystalline salt N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1- yl)-2,4-difluorophenyl)propane-1-sulfonamide with succinic acid, and in particular Form A, can be formed directly from the reaction of N-(3-(5-((1-ethylpiperidin-4-yl) (methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide with succinic acid under proper conditions . Alternatively, the crystalline salt, or Form A thereof, may be prepared by crystallizing the salt and/or recrystallizing the salt from a suitable solvent.

A suitable salt crystallization or salt recrystallization procedure involves dissolving or suspending the salt in a suitable solvent. Can be heated above room temperature, for example, about 30°C, about 40°C, about 50°C, about 60°C, about 70°C, about 80°C, about 90°C, or about 100°C . Heating may be carried out for a period of time, such as about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 40 minutes, about 50 minutes, about 1 hour, about 2 hours, about 3 hours, or about 4 hours. After heating, the solution may be cooled, for example, to a temperature at or below room temperature, for example, about 25°C, about 20°C, about 15°C, about 10°C, about 5°C, or about 0°C. . After such cooling, the reaction mixture may be held for an additional period of time at a lower temperature, for example, for about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 40 minutes, about 50 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 8 hours, about 16 hours, or about 24 hours. During the execution of the steps of crystallization or recrystallization of the salt or its crystalline form, the solution or suspension in which the reaction is carried out may be agitated, for example mixed.

III. Methods of application

The salts and crystalline forms of the present invention are B-RAF kinase inhibitors and are therefore useful in the treatment of pathological disorders (diseases) in which the RAS-RAF-MAPK signaling pathway is activated, in particular cell proliferative diseases such as cancer. Salts and crystalline forms can inhibit cell proliferation, in particular by suppressing access to the DNA synthesis phase. Treated cells are inhibited in the G1 phase of the cell cycle. Therefore, salts and crystalline forms are useful in the treatment of diseases characterized by excessive or abnormal cell proliferation.

Pathological diseases that can be treated with salts and crystalline forms described in the present invention include diseases associated with abnormal expression or activity of RAF kinase. The disease can be cured by administering to the patient, in need of such treatment, a therapeutically effective amount of a crystalline salt, or any of its embodiments, as described here. In some embodiments, the disease is associated with the expression or activity of a mutated B-RAF kinase. In some embodiments, the disease is associated with the expression or activity of a V600E mutated B-RAF kinase. In other embodiments, the disease may be associated with the expression or activity of a mutated B-RAF kinase selected from the following mutants: R461I, I462S, G463E, G463V, G465A, G465E, G465V, G468A, G468E, N580S, E585K, D593V, F594L, G595R, L596V, T598I, V599D, V599E, V599K, V599R, V600K, and A727V.

Pathological diseases that can be treated with the salts and crystalline forms described in the present invention include cancer, infectious diseases, inflammation, and autoimmune diseases.

Pathological diseases that can be treated with the salts and crystalline forms described in the present invention include oncological diseases. Oncological diseases may include neoplasms, and also oncological diseases that are not accompanied by the formation of tumors, such as hematological cancers.

In some embodiments, the cancer is breast cancer, prostate cancer, colon cancer, endometrial cancer, brain cancer, bladder cancer, skin cancer, uterine cancer, ovarian cancer, lung cancer, pancreatic cancer, kidney cancer, stomach cancer, or blood cancer. In some embodiments, the blood cancer is acute myeloid leukemia, chronic myeloid leukemia, B-cell lymphoma, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma, hairy cell leukemia, mantle cell lymphoma, Burkitt's lymphoma, small cell lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma , extranodal marginal zone B cell lymphoma, diffuse large B cell lymphoma of the activated B cell subtype (ABC), or diffuse large B cell lymphoma of the germinal center B cell subtype (GCB). In some embodiments, non-Hodgkin's lymphoma (NHL) is selected from recurrent NHL, resistant NHL, and recurrent follicular NHL.

In some embodiments, the cancer is breast, prostate, colon, endometrial, brain, bladder, skin, uterine, ovarian, lung, pancreatic, kidney, stomach, or blood cancer.

In some embodiments, the cancer is malignant melanoma, thyroid cancer, colorectal cancer, bile duct cancer, prostate cancer, ovarian cancer, or non-small cell lung cancer.

In some embodiments, cancer is associated with RAF kinase expression or activity.

In some embodiments, the cancer is associated with the expression or activity of B-RAF kinase.

In some embodiments, the cancer is associated with the expression or activity of a mutated B-RAF kinase.

In some embodiments, the cancer is associated with the expression or activity of a V600E mutated B-RAF kinase.

In some embodiments, the cancer is associated with the expression or activity of a V600E mutated B-RAF kinase selected from the following mutants: R461I, I462S, G463E, G463V, G465A, G465E, G465V, G468A, G468E, N580S, E585K, D593V, F594L, G595R, L596V, T598I, V599D, V599E, V599K, V599R, V600K, and A727V.

In some embodiments, the cancer is melanoma.

In some embodiments, the cancer is colorectal cancer.

In some embodiments, the cancer is colon cancer.

In some embodiments, the cancer is thyroid cancer.

In some embodiments, the cancer expresses a mutated B-RAF kinase. In some embodiments, the cancer expresses one or more of the following B-RAF kinase mutants: R461I, I462S, G463E, G463V, G465A, G465E, G465V, G468A, G468E, N580S, E585K, D593V, F594L, G595R, L596V, T598I, V599D, V599E, V599K, V599R, V600K, and A727V.

The following cancers can be treated with the compounds according to the invention, without being limited to these: brain tumors such as acoustic neuroma, astrocytomas such as piloid astrocytomas, fibrillar astrocytoma, protoplasmic astrocytoma, gemistocytic astrocytoma, anaplastic astrocytoma and glioblastoma, brain lymphomas, brain metastases, pituitary tract tumor such as HGH (human growth hormone) producing prolactinoma and ACTH (adrenocorticotropic hormone) producing tumor, craniopharyngiomas, medulloblastomas, meningiomas, and oligodendrogliomas; tumors of the nerves (neoplasms), such as, for example, tumors of the autonomic nervous system, such as sympathogonioma, ganglioneuroma, paraganglioma (pheochromocytoma, chromaffinoma), and carotid glomus tumor, tumors of the peripheral nervous system, such as amputation neuroma, neurofibroma, neurinoma (neurilemma, schwannoma) and malignant schwannoma, as well as tumors of the central nervous system, such as brain tumors and bone marrow tumors; intestinal cancer, such as, for example, rectal carcinoma, colon carcinoma, colorectal carcinoma, anal cancer, colon cancer, tumors of the small intestine and duodenum; eyelid tumors such as basalioma or basal cell carcinoma; pancreatic cancer or pancreatic carcinoma; bladder cancer or bladder carcinoma; lung cancer (bronchial carcinoma), such as, for example, small cell bronchial carcinomas (oat cell carcinomas), and non-small cell bronchial carcinomas (NSCLCs) such as squamous epithelial carcinomas, adenocarcinomas, and large cell bronchial carcinomas; breast cancer such as, for example, breast carcinoma such as intraductal carcinoma, mucoid carcinoma, invasive lobular carcinoma, tubular carcinoma, adenoid cystic carcinoma, and papillary carcinoma; non-Hodgkin's lymphoma (NHL), such as, for example, Burkitt's lymphoma, non-Hodgkin's lymphomas (NHL) with low malignant potential, and mycosis fungoides; uterine cancer, or endometrial cancer, or endometrial carcinoma, or endometrial cancer; CUP-syndrome (cancer without an identified primary focus); ovarian cancer or ovarian carcinoma such as mucosal cancer, endometrial cancer or serous cancer; gallbladder cancer; bile duct cancer such as, for example, Klatskin's tumor; testicular cancer, such as seminomas and nonseminomas, for example; lymphoma (lymphosarcoma), such as malignant lymphoma, Hodgkin's disease, non-Hodgkin's lymphomas (NHL), such as chronic lymphocytic leukemia, hairy cell leukemia, immunocytoma, plasmacytoma (multiple myeloma), immunoblastoma, Burkitt's lymphoma, T-cell mycosis fungoides, large cell anaplastic lymphoblastoma and lymphoblastoma; cancer of the larynx, such as, for example, tumors of the vocal cords, supraglottal, glottal and subglottal tumors; bone cancer, such as, for example, osteochondroma, chondroma, chondroblastoma, chondromyxoid fibroma, osteoma, osteoid osteoma, osteoblastoma, eosinophilic granuloma, giant cell tumor, chondrosarcoma, osteosarcoma, Ewing's sarcoma, reticulosarcoma, plasmacytoma, fibrous dysplasia, juvenile bone cysts, and aneurysmal bone cysts ; tumors of the head and neck, such as those of the lips, tongue, floor of the mouth, oral cavity, gums, palate, salivary glands, throat, nasal cavity, paranasal sinuses, larynx and middle ear, for example; liver cancer, such as, for example, hepatocellular carcinoma or hepatocellular carcinoma (HCC); leukemias, for example, such as acute leukemia, such as acute lymphatic/lymphoblastic leukemia (ALL), acute myeloid leukemia (AML); chronic leukemias such as chronic lymphatic leukemia (CLL), chronic myeloid leukemia (CML); gastric cancer or gastric carcinoma, such as, for example, papillary, tubular and mucinous adenocarcinoma, signet cell carcinoma, adenosquamous carcinoma, small cell carcinoma and undifferentiated cancer; melanomas such as, for example, superficial widespread, nodular, lentigo malignant and acral lentiginous melanoma; kidney cancer, such as, for example, renal cell carcinoma or hypernephroma or Gravitz tumor; esophageal cancer or esophageal carcinoma; penile cancer; prostate cancer; cancer of the throat or carcinomas of the pharynx, such as, for example, carcinomas of the nasopharynx, carcinomas of the oropharynx, and carcinomas of the hypopharynx; retinoblastoma, vaginal cancer, or vaginal carcinoma; squamous epithelial carcinomas, adenocarcinomas, pre-invasive carcinomas, malignant melanomas and sarcomas; thyroid carcinomas, such as, for example, papillary, follicular and medullary thyroid carcinomas, as well as anaplastic carcinomas; spinalioma, epidermoid carcinoma and squamous epithelial cancer of the skin; thymomas, urethral cancer and vulvar cancer.

The new salts and crystalline forms can be used for the prevention, short-term or long-term treatment of the aforementioned diseases, optionally also in combination with radiotherapy or other "state of the art" compounds, such as cytostatics or cytotoxins, cell proliferation inhibitors, anti-angiogenic drugs, steroids or antibodies.

The salts and crystalline forms described herein can also be used to treat non-cancer proliferative diseases. Examples of proliferative diseases that may be treated include, but are not limited to, benign soft tissue tumors, bone tumors, brain and spinal cord tumors, eyelid and orbital tumors, granuloma, lipoma, meningioma, multiple endocrine neoplasia, nasal polyps, pituitary tumors , prolactinoma, benign intracranial hypertension, seborrheic keratoses, gastric polyps, thyroid nodules, pancreatic cystitis neoplasms, hemangiomas, vocal cord nodules, polyps and cysts, Castleman's disease, chronic pilonidal disease, dermatofibroma, pilar cyst, pyogenic granuloma, and syndrome juvenile polyposis.

Other diseases that can be treated include viral infections (eg, Epstein-Barr virus, hepatitis B virus, hepatitis C virus, herpes virus, human immunodeficiency virus, human papillomavirus, Kaposi's sarcoma, adenovirus, poxvirus, and other viruses with episomal DNA ). Therefore, salts and crystalline forms can be used to treat diseases and conditions such as herpes simplex infections and reactivations, herpetic cold sores, zoster infections and reactivations, chickenpox, herpes zoster, human papillomavirus, cervical neoplasia, adenovirus infections, including acute respiratory disease; and poxvirus infections such as vaccinia and smallpox, and African swine fever virus. In one specific embodiment, the salts and crystalline forms are indicated for the treatment of papillomavirus infections of the human skin or cervical epithelium.

Other diseases that may be treated include inflammatory conditions and autoimmune diseases. Examples of autoimmune diseases and inflammatory conditions that can be treated include acute, hyperacute or chronic organ transplant rejection, acute gouty arthritis, acute inflammatory reactions (such as acute respiratory distress syndrome and ischemia/reperfusion injury), Addison's disease, agammaglobulinemia, allergic rhinitis, allergy, alopecia, Alzheimer's disease, appendicitis, atherosclerosis, asthma, osteoarthritis, juvenile arthritis, psoriatic arthritis, rheumatoid arthritis, atopic dermatitis, autoimmune alopecia, autoimmune hemolytic and thrombocytopenic conditions, autoimmune hypopituitarism, autoimmune poly glandular syndrome, Behçet's disease, bullous epidermolysis, cholecystitis, chronic idiopathic thrombocytopenic purpura, chronic obstructive pulmonary disease (COPD), cirrhosis, degenerative joint disease, depression, dermatitis, dermatomyositis, eczema, enteritis, encephalitis, gastritis, glomerulonephritis, giant cell arteritis, Goodpasture's syndrome, Guillain-Barré syndrome, gingivitis, Graves' disease, Hashimoto's thyroiditis, hepatitis, hypophysitis, inflammatory bowel disease (Crohn's disease and ulcerative colitis), pelvic inflammatory disease, irritable bowel syndrome, Kawasaki disease, endotoxin-induced shock, meningitis, multiple sclerosis, myocarditis, myasthenia gravis, mushroom mycosis, myositis, nephritis, osteomyelitis, pancreatitis, Parkinson's disease, pericarditis, pernicious anemia, pulmonitis, primary sclerosing cholangitis of the bile ducts, polyarthritis nodosa, psoriasis, retinitis, scleritis, sclerasierma, scleroderma, sinusitis, Sjögren's disease, sepsis, septic shock, solar burn, systemic lupus erythematosus, tissue flap rejection, thyroiditis, type 1 diabetes, Takayasu's arteritis, urethritis, uveitis, vasculitis, vasculitis including giant cell arteritis, vasculitis involving organs such as glomerulonephritis, vitiligo, Waldenström's macroglobulinemia, and Wegener's granulomatosis.

Additional diseases include bacterial, fungal and/or parasitic infections; skin diseases (eg psoriasis); diseases due to hyperplasia, which are characterized by an increase in the number of cells (for example, fibroblasts, hepatocytes, bone and bone marrow cells, cartilage cells, or smooth muscle cells, or epithelial cells (for example, endometrial hyperplasia); bone diseases and cardiovascular diseases (for example, restenosis and hypertrophy).

The salts and crystalline forms described herein are also useful in protecting proliferating cells (eg, hair, intestinal, blood, and undifferentiated cells) from DNA damage caused by radiation, UV exposure, and/or cytotoxic therapy.

Combination Therapies

The salts and crystalline forms described here can be used alone or in combination with other active substances according to the invention, optionally also in combination with other pharmacologically active substances, such as other chemotherapeutic agents.

For the treatment of cancer and other proliferative diseases, the compounds of the invention may be used in combination with chemotherapeutic agents, or with other antiproliferative drugs. Chemotherapeutic agents that may be administered in combination with the salts and crystalline forms described herein for the treatment of cancer and other proliferative diseases include, but are not limited to, hormones, hormone analogs, and antihormonal drugs (e.g., tamoxifen, toremifene, raloxifene, fulvestrant, megestrol acetate , flutamide, nilutamide, bicalutamide, aminoglutethimide, cyproterone acetate, finasteride, buserelin acetate, fludrocortisone, fluoxymesterone, medroxyprogesterone, octreotide), aromatase inhibitors (eg, anastrozole, letrozole, liarozol, vorozol, exemestane, atamestane), LHRH agonists and antagonist ist LHRH ( e.g. goserelin acetate, luprolide), inhibitors of growth factors (growth factors such as "platelet growth factor (PDGF)", "fibroblast growth factor (FGF)", "vascular endothelial growth factor (VEGF)", "epidermal growth factor (EGF)", "insulin-like growth factors (IGF)", "human epidermal growth factor (HER, e.g. HER2, HER3, HER4)", and "hepatocyte growth factor (HGF)"), for example, inhibitors are anti-growth factor antibodies, anti-growth factor receptor antibodies, and tyrosine kinase inhibitors such as cetuximab, gefitinib, imatinib, lapatinib, and trastuzumab); antimetabolites (eg, antifolates such as methotrexate, raltitrexed, pyrimidine analogs such as 5-fluorouracil, capecitabine and gemcitabine, purine and adenosine analogs such as mercaptopurine, thioguanine, cladribine and pentostatin, cytarabine, fludarabine); antitumor antibiotics (eg anthracyclines such as doxorubicin, daunorubicin, epirubicin and idarubicin, mitomycin-C, bleomycin, dactinomycin, plicamycin, streptozocin); platinum derivatives (eg cisplatin, oxaliplatin, carboplatin); alkylating agents (eg estramustine, meclorethamine, melphalan, chlorambucil, busulfan, dacarbazine, cyclophosphamide, ifosfamide, temozolomide, nitrosoureas such as carmustine and lomustine, thiotepa); antimitotic drugs (eg vinca alkaloids such as vinblastine, vindesine, vinorelbine and vincristine; and taxanes such as paclitaxel, docetaxel); tubulin inhibitors; PARP inhibitors, topoisomerase inhibitors (eg epipodophyllotoxins such as etoposide and etopophos, teniposide, amsacrine, topotecan, irinotecan, mitoxantrone), serine/threonine kinase inhibitors (eg PDK 1 inhibitors, B-Raf inhibitors, mTOR inhibitors, mTORC1 inhibitors , PI3K inhibitors, dual mTOR/PI3K inhibitors, STK 33 inhibitors, AKT inhibitors, PLK 1 inhibitors, CDKs inhibitors, Aurora kinase inhibitors), tyrosine kinase inhibitors (eg, PTK2/FAK inhibitors), inhibitors of protein-protein interactions (eg, IAP , Mcl-1, MDM2/MDMX), MEK inhibitors, ERK inhibitors, IGF-1R inhibitors, ErbB receptor inhibitors, rapamycin analogs (eg, everolimus, temsirolimus, ridaphorolimus, sirolimus) and various chemotherapy drugs such as aminophostin, anagrelide, clodronate, filgrastin, interferon, interferon alfa, leucovorin, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate, and porfimer.

Other drugs that can be administered in combination with the salts and crystalline forms described herein include 2-chlorodeoxyadenosine, 2-fluorodeoxycytidine, 2-methoxyestradiol, 2C4, 3-alethine, 131-I-TM-601, 3CPA, 7-ethyl- 10-hydroxycamptothecin, 16-aza-epophilon B, A 105972, A 204197, abarelix, abiraterone, aldesleukin, alemtuzumab, alitretinoin, allopurinol, allovectin-7, altretamine, alvocidib, amonafide, anastrozole, anthrapyrazole, AG-20 37, AP-5280 , apazicuone, apomin, aranose, arglabin, arsenic trioxide, arzoxifene, asparaginase, atamestane, anthrasentan, auristatin PE, AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab), ARRY 162, ARRY 438162, ARRY-300, AR RY- 142886/AZD-6244 (selumetinib), ARRY-704/AZD-8330, AR-12, AR-42, AS-703988, AXL-1717, AZD-8055, AZD-5363, AZD-6244, ARQ-736, ARQ 680, AS-703026 (Primacertib), Avastin, AZD-2014, Azacitidine, Azaepothilone B, Azonafib, BAY-43-9006, BAY 80-6946, BBR-3464, BBR-3576, Bevacizumab, Bexarotene, BEZ-235, Biricodara Dicitrate BCX-1777 BKM-120 Bleocin (afatinib, tomtovok), BIBF 1120 (vargatef), BI 836845, BI 2536, BI 6727, BI 836845, BI 847325, BI 853520, BIIB-022, bleomycin acid, bleomycin A, bleomycin B, brivanib, bryostatin-1 , bortezomib , brostallicin, bisulfan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol, calusterone, canertinib, canfosfamide, capecitabine, carboplatin, carboxyphthalatoplatin, carmustine, CCI-779, CC-115, CC-223, CEP- 701, CEP-751, CBT-1, cefixime, ceflatonin, ceftriaxone, celecoxib, celmoleukin, cematodin, cetuximab, chlorambucil, CH4987655/RO-4987655, chlortrianisene, cilengitide, cisplatin, CDA-II, CDC-394, CKD-602, CKI-27, cladribine, clofarabine, colchicine, combretastatin A4, COT inhibitors, CHS-828, CH-5132799, CLL-Thera, CMT-3 cryptophycin 52, CTP-37, CTLA-4 monoclonal antibodies, CP-461, CV- 247, cyanomorpholinodoxorubicin, cyclophosphamide, cyclosporine, cyanomorpholinodoxorubicin, D 24851, dacarbazine, dactinomycin, dalteparin sodium, dasatinib, daunorubicin, decitabine, denileukin, denileukin diftitox, deoxorubicin, deoxyrubicin, deoxycoformycin, depsipeptide , deoxyepothilone B, dexamethasone, dexrazoxane, dexrazoxanet, diethylstilbestrol, diftomothecan, didox, DMDC, dolastatin 10, docetaxel, doranidazole, doxorubicin, dromostanolone propionate, DS-7423, E7010, E-6201, eculizumab, edatrexate, edotreotide, efaproxiral, eflornithine, EGFR inhibitors, EKB-569, EKB-509, enzastaurin , elzamitrucin, epirubicin, epothilone B, epratuzumab, ER-86526, erlotinib, estramustine, ET-18-OCH3, ethinylcytidine, ethinylestradiol, etoposide phosphate, etoposide, exatecan, exatecan mesylate, exemestane, exisulind, fentanyl citrate, fenretinide, figit umumab, filgrastin , floxuridine, fludarabine, folic acid, fluorouracil, FOLFOX, FOLFOX4, FOLFIRI, formestane, fotemustine, fulvestrant, galarubicin, gallium maltolate, gefinitib, gemcitabine, gemtuzumab, gemtuzumab ozogamicin, gimatecan, glufosfamide, GCS-IOO , GDC-0623, GDC- 0941 (pictrelisib), GDC-0980, GDC-0032, GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GPX-100, gp100-peptide vaccines, GSK-5126766, GSK-690693, GSK-1120212 (trametinib), GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A, GSK-2334470, GSK-2110183, GSK-2141795, GW2016, goserelin acetate, granisetron, herceptin, hexamethylmelamine, histamine, histrelin acetate, homochar ringtonin, homoharringtonin , hydroxyurea, hydroxyprogesterone caproate, ibandronate, ibritumomab, ibritumomab tiuxetan, idarubicin, idatrexate, idenestrol, IDN-5109, ifosfamide, IGF-1R inhibitors, IMC-1C11, IMC-A12 (cixutumumab), imatinib mesylate, immunol, indisulam, interfer he is alpha -2a, interferon alfa-2b, pegylated interferon alfa-2b, interleukin-2, INK-1117, INK-128, INSM-18, ionafarnib, ipilimumab, iproplatin, irinotecan, irofulven, isohomogalichondrin-B, isoflavone, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088, Conjugated Estrogens, Cachalide F, Ketoconazole, KW-2170, KW-2450, Lapatinib Ditosylate, Leflunomide, Lenalidomide, Lenograstim, Letrozole, Leucovorin, Leuprolide, Leuprolide Acetate, Leuporelin, Levamisole, lexidronam, LGD-1550, linezolid, lobaplatin, lutetium texafirin, lometrexol, lomustine, losoxantrone, LU 223651, lurtothecan, LY-S6AKT1, LY-2780301, mafosfamide, marimastat, mechlorethamine, megestrol acetate, MEK inhibitors, M EK-162, melphalan, меркаптопурин, метотрексат, метоксзален, метилтестостерон, метилпреднизолон, MEDI-573, MEN-10755, MDX-H210, MDX-447, MDX-1379, MGV, мидостаурин, минодронную кислоту, митомицин, митомицин C, митотан, митоксантрон, мивобулин, MK- 2206, MK-0646 (dalotuzumab), MLN518, gadolinium motexafine, MS-209, MS-275, MX6, nandrolone phenylpropionate, nelarabine, neridronate, neratinib, Nexavar, neovastat, nilotinib, nimesulide, nitroglycerin, nofetumomab, nolatrexed, norelin, N -acetylcysteine, N-benzylguanine, oblimersen, omeprazole, oncophage, oncoVEX ^GM-CSF , ormiplatin, orthotaxel, oxaliplatin, anti-OX44 antibodies, OSI-027, OSI-906 (linsitinib), anti-4-1BB antibodies, oxantrazole, estrogen, paclitaxel , pamidronate, panitumumab, patupilone, pegfilgrastim, PCK-3145, pegaspargas, pegfilgrastim, PBI-1402, PBI-05204, PDO325901, PD-1 antibodies, PEG-paxitaxel, albumin-stabilized paxitaxel, PEP-005, PF-05197281 , PF -05212384, PF-04691502, PHT-427, P-04, PKC412, P54, PI-88, Pelitinib, Pemetrexed, Pemetrexed Disodium, Pentostatin, Pentrix, Perifosine, Peryl Alcohol, Pertuzumab, PI3K Inhibitors, PI3K/mTOR Inhibitors, PG -TXL, PG2, PLX-4032/RO-5185426 (vemurafenib), PLX-3603/RO-5212054, PT-100, PWT-33597, PX-866, picoplatin, pipobroman, pivaloyloxymethylbutyrate, pixantrone, phenoxodiol O, PKI166, plevitrexed , plicamycin, polyprenic acid, porfiromycin, prednisone, prednisolone, procarbazine, quinacrine, gynamed, ginupristin, R115777, RAF-265, ramosetron, ranpirnase, rasburicua, RDEA-119/BAY 869766, RDEA-436, rebeccamycin analogs, receptor inhibitors tyrosine kinase ( RTK), revimid, RG-7167, RG-7304, RG-7421, RG-7321, RG 7440, rizoxin, rhu-MAb, rinfabate, risedronate, rituximab, robatumumab, rofecoxib, RO-31-7453, RO-5126766, RO-5068760, RPR 109881A, rubidazon, rubinecan, R-flurbiprofen, RX-0201, S-9788, sabarubicin, SAHA, sargramostim, satraplatin, SB 408075, Se-015/Ve-015, SU5416, SU6668, SDX-101, semustine, seocalcitol, SM-11355, SN-38, SN-4071, SR-27897, SR-31747, SR-13668, SRL-172, sorafenib, spiroplatin, squalamine, streptozocin, suberanilohydroxamic acid, sunitinib, sunitinib maleate, sutent, T 900607, T 138067, TAK-733, TAS-103, tacedinaline, thalaporfin, tamoxifen, Tarceva, tariguitar, tasisulam, taxotere, taxoprexin, tazarotene, tegafur, temozolamide, teniposide, tesmilifen, testosterone, testosterone propionate, tesmilifen, testolactone, tetraplatin , tetrodotoxin, tezacitabine, thalidomide, teralux, terarubicin, thioguanine, thiotepa, thymalfacin, thiazofurin, tipifarnib, tirapazamine, tocladesin, tomudex, topotecan, toremifene, toremofin, tositumomab, trabectedin, TransMID-107, trans-retinoic acid, trastuzumab, tremelimumab, tretinoin, triacetyluridine, thiapine, triciribine, trimetrexate, TLK-286TXD 258, tikerb/tyverb, urocidin, uracil mustard, valrubicin, vatalanib, vinblastine, vincristine, vinflunine, vinorelbine, virulizin, WX-UK1, WX-554, vestibix, vorinostat, XELOX 4054, ZD-0473, ZD-6126, ZD-9331, ZDI839, ZSTK-474, zoledronate and zosuguidar.

The compounds of the invention may also be used in combination with medical therapies such as surgery or radiotherapy, for example with gamma radiation, neutron beam radiotherapy, electron beam radiotherapy, proton beam therapy, brachytherapy, and systemic radioactive isotopes.

For the treatment of autoimmune diseases and inflammatory processes, the compound of the invention may be administered in combination with a corticosteroid such as triamcinolone, dexamethasone, fluocinolone, cortisone, prednisolone or flumetholone.

For the treatment of autoimmune diseases and inflammatory conditions, a compound of the invention may be administered in combination with an immunosuppressant such as fluocinolone acetonide (Retisert®), rimexolone (AL-2178, Vexol, Alcon), or cyclosporine (Restasis®).

For the treatment of autoimmune diseases and inflammatory processes, the compound of the invention may be administered in combination with one or more additional drugs selected from Dehydrex™ (Holles Labs), Cyvamide (Opko), sodium hyaluronate (Vismed, Lantibio/TRB Chemedia), cyclosporine (ST- 603, Sirion Therapeutics), ARG101(T) (testosterone, Argentis), AGR1012(P) (Argentis), sodium ecabeta (Senju-Ista), gefarnate (Santen), 15-(s)-hydroxyeicosantetraenoic acid (15(S) -HETE), cevilemin, doxycycline (ALTY-0501, Alacrity), minocycline, iDestrin™ (NP50301, Nascent Pharmaceuticals), cyclosporine A (Nova22007, Novagali), oxytetracycline (Duramycin, MOLI1901, Lantibio), CF101 (2S, 3S, 4R , 5R)-3,4-dihydroxy-5-[6-[(3-iodophenyl)methylamino]purin-9-yl]-N-methyl-oxolane-2-carbamyl, Can-Fite Biopharma), voclosporin (LX212 or LX214, Lux Biosciences), ARG103 (Agentis), RX-10045 (synthetic analog of resolvin, Resolvyx), DYN15 (Dyanmis Therapeutics), rivoglitazone (DE011, Daiichi Sanko), TB4 (RegeneRx), OPH-01 (Ophtalmis Monaco), PCS101 (Pericor Science), REV1-31 (Evolutec), Lacritin (Senju), rebamipide (Otsuka-Novartis), OT-551 (Othera), PAI-2 (University of Pennsylvania and Temple University), pilocarpine, tacrolimus, pimecrolimus (AMS981 , Novartis), loteprednol etabonate, rituximab, tetrasodium diquafazole (INS365, Inspire), KLS-0611 (Kissei Pharmaceuticals), dehydroepiandrosterone, anakinra, efalizumab, sodium mycophenolate, etanercept (Embrel®), hydroxychloroquine, NGX267 (TorreyPines Therapeutics), or thali domida .

In some embodiments, a compound of the invention may be administered in combination with one or more drugs selected from antibiotics, antivirals, antifungals, anesthetics, anti-inflammatory agents, including steroidal and non-steroidal anti-inflammatory drugs, and antiallergic agents. Examples of suitable medicaments include aminoglycosides such as amikacin, gentamicin, tobramycin, streptomycin, netilmicin and kanamycin; fluoroquinolones such as ciprofloxacin, norfloxacin, ofloxacin, trovafloxacin, lomefloxacin, levofloxacin and enoxacin; naphthyridine; sulfonamides; polymyxin; chloramphenicol; neomycin; paramomycin; colistimethate; bacitracin; vancomycin; tetracyclines; rifampin and its derivatives ("rifampins"); cycloserine; beta lactams; cephalosporins; amphotericins; fluconazole; flucytosine; natamycin; miconazole; ketoconazole; corticosteroids; diclofenac; flurbiprofen; ketorolac; Suprofen; cromolyn; lodoxamide; levocabastine; naphazoline; antazolin; pheniramine; or an azalide antibiotic.

One or more additional pharmaceutical preparations may be administered to the patient simultaneously or sequentially.

IV. Preparation, dosage forms and administration

When used as pharmaceuticals, the salts and crystalline forms described herein may be administered in the form of pharmaceutical compositions. These compositions may be formulated in a manner well known in pharmaceutical technology, and may be administered in a variety of ways, depending on whether local or systemic treatment is desired and on the area being treated. Administration can be topical (including transdermal, epidermal, ocular, and mucosal, including intranasal, vaginal, and rectal administration), pulmonary (e.g., inhalation or inhalation of powders or aerosols, including by nebulizer), oral, or parenteral. Parenteral administration includes intravenous, intra-arterial, subcutaneous, intraperitoneal, intramuscular administration, or by injection or infusion; or intracranial, eg intrathecal or intraventricular, administration. Parenteral administration may be in the form of a single bolus dose, or may be, for example, a continuous perfusion infusion. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, aerosols, solutions and powders. Conventional pharmaceutical carriers, aqueous, powder or oil based, thickeners and the like may be necessary or desirable. The content of the pharmaceutically active compound should be in the range of 0.1 to 90 wt.%, preferably 0.5 to 50 wt.%, of the composition as a whole, that is, in an amount that is sufficient to achieve the following dosage range.

This invention also includes pharmaceutical compositions which contain, as the active ingredient, the salts and crystalline forms described herein, in combination with one or more pharmaceutically acceptable carriers (excipients). In some embodiments, the composition is suitable for topical use. In preparing the compositions of the invention, the active ingredient is usually mixed with an excipient, diluted with an excipient, or enclosed within such a carrier in the form of, for example, a capsule, small pouch, paper, or other container. When an excipient serves as a diluent, it may be a solid, semi-solid, or liquid material that acts as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions may be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (both solid and liquid), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders.

In formulating the active compound in salt or crystalline form, as described herein, may be ground into particles of the appropriate size before being combined with other ingredients. If the salt or crystalline form is substantially insoluble, it can be ground to a particle size of less than 200 mesh. If the salt or crystalline form is substantially water soluble, the particle size can be adjusted by milling to provide a substantially uniform distribution in the composition, eg about 40 mesh.

The salts and crystalline forms described herein can be milled using known milling techniques, such as wet milling, to obtain particle sizes suitable for tablet formation and other types of compositions. Finely divided (eg, to nanoparticles) preparations of the salts and crystalline forms described herein can be obtained by methods known in the art, for example, see International Patent Application No. WO 2002/000196.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum arabic, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. The compositions may further include: lubricants such as talc, magnesium stearate and mineral oil; wetting agents; emulsifiers and suspending agents; preservatives such as methyl and propyl hydroxybenzoates; sweeteners; and flavor additives. The salts and crystalline forms described herein may be formulated to provide rapid, sustained or delayed release of the active ingredient after administration to a patient using procedures known in the art.

Suitable tablets can be obtained, for example, by mixing the active substance(s) with known excipients, for example, inert diluents such as calcium carbonate, calcium phosphate or lactose, disintegrants such as corn starch or alginic acid, binders , such as starch or gelatin, lubricants such as magnesium stearate or talc, and/or delayed release agents such as carboxymethyl cellulose, cellulose acetate phthalate, or polyvinyl acetate. Tablets may also include multiple layers.

The compositions may be formulated in unit dosage form, with each dosage form containing from about 5 to about 1000 mg (1 g), more typically from about 100 mg to about 500 mg, of the active ingredient. The term "unit dosage form" means physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined amount of active material calculated to produce the desired therapeutic effect in association with a suitable pharmaceutical excipient.

In some embodiments, the compositions according to the invention contain from about 5 mg to about 50 mg of the active ingredient. One of ordinary skill in the art will appreciate that compounds or compositions containing from about 5 mg to about 10 mg, from about 10 mg to about 15 mg, from about 15 mg to about 20 mg, from about 20 mg to about 25 mg, about 25 mg to about 30 mg, about 30 mg to about 35 mg, about 35 mg to about 40 mg, about 40 mg to about 45 mg, or about 45 mg to about 50 mg active ingredient. The doses described may be a given amount of a particular salt or crystal form, or an amount of a particular salt or crystal form that produces a given dose of the free base compound (namely, N-(3-(5-((1-ethylpiperidine-4- yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propan-1-sulfonamide).

In some embodiments, the compositions according to the invention contain from about 50 mg to about 500 mg of the active ingredient. One of ordinary skill in the art will appreciate that compounds or compositions containing from about 50 mg to about 100 mg, from about 100 mg to about 150 mg, from about 150 mg to about 200 mg, from about 200 mg to about 250 mg, about 250 mg to about 300 mg, about 350 mg to about 400 mg, or about 450 mg to about 500 mg of the active ingredient. The doses described may be a given amount of a particular salt or crystal form, or an amount of a particular salt or crystal form that produces a given dose of the free base compound (namely, N-(3-(5-((1-ethylpiperidine-4- yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propan-1-sulfonamide).

In some embodiments, compositions according to the invention contain from about 500 mg to about 1000 mg of the active ingredient. One of ordinary skill in the art will appreciate that compounds or compositions containing from about 500 mg to about 550 mg, from about 550 mg to about 600 mg, from about 600 mg to about 650 mg, from about 650 mg to about 700 mg, about 700 mg to about 750 mg, about 750 mg to about 800 mg, about 800 mg to about 850 mg, about 850 mg to about 900 mg, about 900 mg to about 950 mg, or from about 950 mg to about 1000 mg of the active ingredient. The doses described may be a given amount of a particular salt or crystal form, or an amount of a particular salt or crystal form that produces a given dose of the free base compound (namely, N-(3-(5-((1-ethylpiperidine-4- yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propan-1-sulfonamide).

The predetermined doses may be for one dose per day, or, if necessary, for several doses per day, for example, twice, thrice, or four times a day.

The active compound may be effective over a wide dosage range and is generally administered in a pharmaceutically effective amount. However, it will be understood that the actual dose administered will generally be determined by the attending physician, according to relevant circumstances, including the condition being treated, the route of administration chosen, the actual compound administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.

To obtain solid compositions such as tablets, the main active ingredient is mixed with a pharmaceutical excipient to form a solid preformulated composition containing a homogeneous mixture of salt or crystalline form, as described here. When these pre-formulated compositions are described as uniform, the active ingredient is usually uniformly dispersed throughout the composition so that the composition can be easily subdivided into equally effective unit dosage forms such as tablets, pills and capsules. This solid preformulated composition is then divided into unit dosage forms of the type described above, containing, for example, from about 0.1 to about 1000 mg of the active ingredient.

Tablets or pills containing the salts and crystalline forms described herein may be coated or otherwise compounded to provide a dosage form that provides the benefit of a delayed release. Coated tablets can be obtained by coating the cores, formed similarly to tablets, with substances commonly used for coating tablets, for example collidone or shellac, gum arabic, talc, titanium dioxide or sugar. To provide delayed release or prevent incompatibilities, the core may also be composed of multiple layers. Likewise, the coating on a tablet may consist of several layers to achieve delayed release, possibly using the above mentioned tablet excipients.

For example, a tablet or pill may include an inner dose component and an outer dose component, the latter in the form of a shell on top of the former. The two components may be separated by an enteric layer which serves to resist disintegration in the stomach and allows the inner component to pass unaffected to the duodenum, or whose release is delayed. A variety of materials can be used for such enteric layers or coatings, and such materials include certain polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

Liquid forms into which the salts, crystalline forms and compositions of the invention may be incorporated for oral or injectable use include aqueous solutions, suitable flavored syrups, aqueous or oily suspensions, and flavored emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and similar pharmaceutical carriers. Although liquid compositions in which the salt is dissolved will not normally contain the salt in crystalline form, the salts and crystalline forms described herein will nevertheless be useful in preparing liquid compositions, for example, by dissolving the salt or crystalline form thereof in a suitable medium.

Solutions for injection and infusion can be prepared in the usual way, for example by the addition of isotonic solutions, preservatives such as para-hydroxybenzoates or stabilizers such as alkali metal salts of ethylenediaminetetraacetic acid, optionally using emulsifiers and/or dispersants, if water is used as a diluent, for example, organic solvents may optionally be used as solvating additives or to promote dissolution, and transferred into vials or ampoules for injection or bottles for infusion.

Capsules containing one or more active substances or combinations of active substances can be prepared, for example, by mixing the active substances with inert carriers such as lactose or sorbitol and encapsulating them in gelatin capsules.

Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents or mixtures thereof, and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, the compositions are administered into the oral or nasal airways for local or systemic action. The compositions can be sprayed using inert gases. The nebulized solutions may be inhaled directly from a nebulization device, or the nebulization device may be attached to a face mask, a canopy, or a pulsed pressure breathing apparatus. Solution, suspension, or powder compositions may be administered via the mouth or nose from devices that properly deliver the composition.

Compositions for external use may contain one or more conventional carriers. In some embodiments, ointments may contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white petrolatum, and the like. Carrier formulations in creams may be based on water in combination with glycerol and one or many other components such as glycerol monostearate, polyethylene glycol (PEG)-glycerol monostearate and cetyl stearyl alcohol. Gels can be prepared using isopropyl alcohol and water, usually in combination with other ingredients such as glycerol, hydroxyethyl cellulose, and the like. In some embodiments, topical formulations contain at least about 0.1 wt.%, at least about 0.25 wt.%, at least about 0.5 wt.%, at least about 1 wt.% , at least about 2 wt.%, or at least about 5 wt.% of the compounds according to the invention. Formulations for external use may be appropriately packaged in tubes, for example 100 g, which are optionally accompanied by instructions for the treatment of the selected indication, for example psoriasis or other skin disease.

The amount of salt or crystalline form, or composition thereof, administered to a patient will vary depending on what is being administered, the purpose of administration such as prophylaxis or therapy, the condition of the patient, the route of administration, and the like. In therapeutic applications, the compositions may be administered to a patient already in a disease state in an amount sufficient to cure or at least partially eliminate the symptoms of the disease and its complications. Effective doses will depend on the condition of the disease being treated, as well as in the judgment of the attending physician, depending on such factors as the severity of the disease, the age, weight and general condition of the patient, and the like.

The therapeutic dosage of a compound of the present invention may vary according to, for example, the particular application being treated, the route of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of a compound of the invention in a pharmaceutical composition may vary depending on a number of factors including dosage, chemical characteristics (eg hydrophobicity), and route of administration. For example, the compounds of the invention may be provided in an aqueous physiological buffer solution containing from about 0.1% to about 10% (w/v) of the compound for parenteral administration. Some typical dosage ranges are from about 1 μg/kg to about 1 g/kg of body weight per day. In some embodiments, the dosage range is from about 0.01 mg/kg to about 100 mg/kg of body weight per day. Rather, the dose will depend on such variables as the type and extent of the disease or disorder, the general health of the individual patient, the biological efficacy of the compound chosen, the formulation of the excipient, and the route of administration. Effective doses can be extrapolated from dose-response curves derived from in vitro experiments or animal model test systems.

Suitable doses for administration to a human are expected to be in the range of about 50 mg to about 2000 mg, for example, about 100 mg to about 2000 mg, about 200 mg to about 2000 mg, about 400 mg to about 2000 mg, from about 600 mg to about 2000 mg, about 800 mg to about 2000 mg, about 1000 mg to about 2000 mg, about 1200 mg to about 2000 mg, about 1400 mg to about 2000 mg, about 1500 mg to about 2000 mg, about 1600 mg to about 2000 mg, about 1800 mg to about 2000 mg, about 50 mg to about 1500 mg, about 100 mg to about 1500 mg, about 200 mg to about 1500 mg, about 400 mg to about 1500 mg, about 600 mg to about 1500 mg, about 800 mg to about 1500 mg, about 1000 mg to about 1500 mg, about 1200 mg to about 1500 mg, about 1250 mg to about 1500 mg, about 50 mg to about 1000 mg, about 100 mg to about 1000 mg, about 200 mg to about 1000 mg, about 400 mg to about 1000 mg, about 600 mg to about 1000 mg, about 800 mg to about 1000 mg, about 50 mg to about 500 mg, about 100 mg to about 500 mg, about 200 mg to about 500 mg, about 400 mg to about 500 mg, or a dose of about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 600 mg, about 700 mg, about 750 mg, about 800 mg, about 900 mg, about 1000 mg, about 1200 mg, about 1400 mg, about 1500 mg, about 1600 mg, about 1800 mg or about 2000 mg. The proper dose, as given, may be that of the salt itself, producing the amount of compound as the free base (namely, N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-( pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide).

Compositions of the invention may further include one or more additional pharmaceuticals, such as a chemotherapeutic drug, steroid, anti-inflammatory compound, or immunosuppressant, examples of which are listed here below.

The following composition examples illustrate the present invention without limiting its scope:

Examples of pharmaceutical compositions

A)

Pills Per pill Salt or crystalline form of the compound 100 mg Lactose 140 mg Corn starch 240 mg Polyvinylpyrrolidone 15 mg magnesium stearate 5 mg 500 mg

The finely divided active substance, lactose and some cornstarch are mixed together. The mixture is screened, then moistened with a solution of polyvinylpyrrolidone in water, kneaded, wet granulated and dried. The granules, remaining cornstarch and magnesium stearate are screened and blended together. The mixture is compressed to form tablets of the proper shape and size.

IN)

Pills Per pill Salt or crystalline form of the compound 80 mg Lactose 55 mg Corn starch 190 mg Microcrystalline cellulose 35 mg Polyvinylpyrrolidone 15 mg Sodium salt of carboxymethyl starch 23 mg magnesium stearate 2 mg 400 mg

The finely divided active substance, some cornstarch, lactose, microcrystalline cellulose and polyvinylpyrrolidone are mixed together, the mixture is sieved and treated with the rest of the cornstarch and water to form a granulate which is dried and sieved. The sodium carboxymethyl starch and magnesium stearate are added and mixed and the mixture is compressed to form tablets of the proper size.

V. Kits

The present invention also includes pharmaceutical kits useful, for example, for the treatment or prevention of B-RAF related diseases or disorders such as cancer, which include one or more containers containing a pharmaceutical composition containing a therapeutically effective amount of a salt or crystalline form as herein described. In addition, such kits, if desired, may include one or many of the various components of a standard pharmaceutical kit, such as containers with one or more pharmaceutically acceptable carriers, additional containers, etc., as will be readily understood by a person skilled in the art. this area of technology. The kit may also include instructions, both in the form of tabs and in the form of labels, indicating the amount of components to be administered, guidelines for administration, and/or instructions for mixing the components.

The invention will be described in much greater detail by way of specific examples. The following examples are presented for illustrative purposes and are not intended to limit the invention in any way. Those skilled in the art will readily understand the variety of non-critical parameters that can be changed or modified to achieve substantially the same results.

EXAMPLES

General Methods

In the examples below, unless otherwise noted, X-ray powder diffractometry analysis was performed on a STOE STADI powder diffractometer in transmission mode equipped with a position sensitive detector (PSD) and a Cu anode as an X-ray source with monochromatic CuKα1 radiation. (λ=1.54056 Å, 40 kV, 40 mA). The general measurement conditions were as follows:

- starting angle - 3°

- end angle - 40°

- sample - 0.02 deg.

- scanning speed - 10 sec/step.

Differential Scanning Calorimetry (DSC) measurement was performed on a TA Instrument Differential Scanning Calorimetry, model Q2000, using a sample size of 8-10 mg. General experimental conditions were as follows: 25-350°C at 10°C/minute.

Thermogravimetric analysis (TGA) was performed on a TA Instrument Thermogravimetric Analyzer, model Q5000, under the following conditions: temperature rise at a rate of 10°C/minute to 350°C.

Dynamic vapor sorption (DVS) was measured using an IGAsorp Water Sorption Analyzer from Hiden Isochema. Adsorption and desorption isotherms were recorded at 25°C with 10% stepwise RH intervals in the range from 0% relative humidity to 90% relative humidity.

With respect to the HPLC mass spectrometry/UV spectroscopy method, retention time/MS-ESI ⁺ parameters for characterizing exemplary samples according to the invention were obtained using an HPLC-MS (high performance liquid chromatography with mass detector) instrument. Compounds that wash out at the peak of the injection are assigned a retention time t _Ret. =0.00. The methods described below in the Examples are as follows.

Method A HPLC-MS

HPLC: Agilent 1100 Series

MS: Agilent LC/MSD SL

Column: Waters, XBridge™ C18, 5 µm, 2.1 x 50 mm

Eluent: A: H ₂ O (5 mM (NH ₄ ) ₂ CO ₃ , 19 mM NH ₃ )

B: grade acetonitrile for HPLC

Detection: MS: positive and negative mode

Mass range: 120-800 m/z

Flow rate: 1.20 ml/min

Column temperature: room (rt)

Gradient: 0.00 min, 5% B

0.00-1.25 min, 5%→95% B

1.25-2.00 min, 95% B

Method B HPLC-MS

HPLC: Agilent 1100 Series

MS: Agilent LC/MSD SL

Column: Waters, XBridge™ C18, 2.5 µm, 2.1×20 mm

Eluent: A: H ₂ O (0.1% NH ₃ )

B: Grade acetonitrile for HPLC

Detection: MS: positive and negative mode

Mass range: 120-800 m/z

Flow rate: 1.00 ml/min

Column temperature: 60°C

Gradient: 0.00 min 5% B

0.00-2.50 min, 5%→95% B

2.50-2.80 min, 95% B

Method C HPLC-MS

HPLC: Agilent 1100 Series

MS: Agilent LC/MSD SL

Column: Waters, Sunfire™ C18, 5 µm, 2.1 x 50 mm

Eluent: A: H ₂ O (0.2% HCOOH)

B: grade acetonitrile for HPLC (0.2% HCOOH)

Detection: MS: positive and negative mode

Mass range: 120-800 m/z

Flow rate: 1.20 ml/min

Column temperature: rt

Gradient: 0.00 min, 5% B

0.00-1.50 min, 5%→95% B

1.50-2.00 min, 95% B

Equilibrium solubility measurements

Saturated solutions are prepared in multiwell plates by adding the required volume of the selected aqueous medium (typically in the range of 0.25-1.5 ml) to each well that contains a known amount of drug solid (typically in the range of 0.5-5.0 mg). The wells are shaken or mixed for a predetermined period of time (typically in the range of 2-24 hours) and then the contents are filtered using membrane filters (typically 0.45 µm PTFE filters). Absorption on the filter is avoided by releasing the first few drops of the filtrate. The amount of dissolved drug is determined by UV spectroscopy. In addition, the pH value of the aqueous saturated solution is measured using a glass electrode pH meter.

Measurements of intrinsic dissolution rate

N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl)propane-1-sulfonamide was determined in an aqueous medium covering the pH range of 1.1-7.4 using the rotating disk method, which maintains a constant surface area.

The drug substance (5 mg) was pressed with a force of 356.1 Newtons for 60 seconds to form a disk. These discs were mounted in sample holders which were inserted into a miniaturized dissolution tester. The dissolution medium was stirred at a speed of 200 rpm at a temperature of 37°C. Samples were automatically removed every other minute from the dissolution tank and evaluated by UV spectrophotometry.

Preparation of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl )-2,4-difluorophenyl) propane-1-sulfonamide (BI 882370)

Step 1 4-(6-methyl-5-nitropyridin-2-yl)-piperazine-1-carboxylic acid tert-butyl ester (3)

DIPEA (62.82 ml, 0.435 mol) is added to a solution of 6-chloro-3-nitro-2-methylpyridine (1) (50 g, 290 mmol) and N-Boc-piperazine (2) (53.95 g, 290 mmol) in anhydrous MeCN (200 ml) and stirred for 4 hours at 50°C. After completion of the reaction, the reaction mixture was diluted with acetonitrile (MeCN) and water, and stirred for 30 minutes. The precipitated product is isolated by filtration, washed with water and the solid is dried in vacuo.

Step 2 4-[6-((E)-2-dimethylaminovinyl)-5-nitropyridin-2-yl]-piperazine-1-carboxylic acid tert-butyl ester (4)

To a stirred solution of 4-(6-methyl-5-nitropyridin-2-yl)-piperazine-1-carboxylic acid tert-butyl ester (3) (13 g, 40.3 mmol) in DMF (35 ml) was added dimethyl acetal N,N-dimethylformamide (14.47 g, 121 mmol) and stirred under argon for 36 hours at 90°C. An additional 1.5 equivalents of N,N-dimethylformamide dimethylacetal is added and stirred for 12 hours at 90°C. The reaction mixture was poured into water and extracted with dichloromethane (DCM). The combined organic layers are washed with water, dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The residue is used without further purification for the next step.

Step 3 4-(1H-pyrrolo[3,2-b]pyridin-5-yl)piperazine-1-carboxylic acid tert-butyl ester (5)

4-[6-((E)-2-dimethylaminovinyl)-5-nitropyridin-2-yl]-piperazine-1-carboxylic acid tert-butyl ester (36.4 g, 96 mmol) was taken up in MeOH, Pd was added /C (0.56 g, 10%), and the mixture is subjected to hydrogenation in an autoclave at a pressure of 60 psi (0.41 MPa) for 16 hours. The reaction mixture is filtered and concentrated under reduced pressure. The residue was purified by column chromatography under NP MPLC conditions. The product containing fractions of compound (5) (HPLC-MS, method B: t _Ret. =1.55 min; MS (M+H) ⁺ =303) are combined and evaporated in vacuo.

Step 4. N-(3-Amino-2,6-difluorophenyl)acetamide (7)

Compound (6) (55.0 g, 254 mmol) was taken up in MeOH (1.0 L). Add Pd/C (10.0 g, 10%), and the mixture is subjected to hydrogenation in an autoclave at a pressure of 200 psi (1.38 MPa) for 3 hours. The reaction mixture is filtered and concentrated under reduced pressure. The residue is purified under NP-MPLC conditions on silica gel using DCM/MeOH (96:4) as eluent. The product containing the aniline intermediate fractions (HPLC-MS, method B: t _Ret. =0.25 min; MS(MH) ^- =185) was combined and evaporated.

Step 5. N-(2,6-Difluoro-3-propylsulfonamido)phenyl)acetamide (9)

To the aniline intermediate (35.0 g, 188 mmol) in DCM (100 ml) were added pyridine (6.6 ml, 75 mmol) and n-propanesulfonyl chloride (8) (29.5 ml, 263 mmol) and the mixture was stirred at room temperature (rt) for 16 hours. The reaction mixture was diluted with EtOAc (200 mL), washed with H ₂ O and HCl (aq. 1N) and the layers were separated, dried over MgSO ₄ , and evaporated to give the sulfonamide (9), which was used without further purification.

Step 6. N-(3-Amino-2,4-difluorophenyl)propane-1-sulfonamide (10)

Sulfonated aniline (9) (38.0 g, 130 mmol) was taken up in EtOH (250 ml), H ₂ O (200 ml) and concentrated hydrochloric acid (200 ml) were added and heated to 80° C. for 2 hours. The reaction mixture was concentrated under reduced pressure, aqueous NaOH (4N) was added until pH=6 was reached and the mixture was extracted twice with DCM. The combined organic layer was washed with brine, dried over MgSO ₄ , filtered and evaporated to give deacylated aniline (10) (HPLC-MS, method B: t _Ret. =0.22 min; MS(MH) ^- =249) as hydrochloride which was used without further purification.

Step 7. N-(2,4-difluoro-3-iodophenyl)propane-1-sulfonamide (11)

Compound (10) hydrochloride was taken up in DCM and extracted with NaHCO ₃ solution. The organic layer is dried over MgSO ₄ , filtered and evaporated. NaNO ₂ (1.96 g, 28.4 mmol) was added in small portions to the free base (10) (3.55 g, 14.21 mmol) in TFA (80 ml) at 0°C and the mixture was stirred for 30 minutes. KI (23.83 g, 142 mmol) is added and stirring is continued for an additional 15 minutes. The reaction mixture was diluted with ether (Et ₂ O) and stirred for 1 hour. A solution of Na ₂ S ₂ O ₃ (semi-conc.) was added and the mixture was extracted three times with ether (Et ₂ O). The combined organic layer is dried over MgSO ₄ , filtered and concentrated in vacuo. The residue was purified by column chromatography under NP-MPLC conditions. The product containing fractions of compound (11) (HPLC-MS, method A: t _Ret. =1.58 min; MS (MH) ^- =360) are combined and evaporated in vacuo.

Step 8 4-((1-(2,6-difluoro-3-(propylsulfonamido)phenyl)-1H-pyrrolo[3,2-b]pyridin-5-yl)(methyl)amino) tert-butyl ester piperidine-1-carboxylic acid (12)

1H-Pyrrolo[3,2-b]pyridine (5) (10.0 g, 30.27 mmol), sulfonamide (11) (16.4 g, 45.4 mmol), CuI (576 mg, 3.03 mmol), trans-(1R,2R)-N, N'-bismethyl-1,2-cyclohexanediamine (1.91 ml, 12.1 mmol) and Cs ₂ CO ₃ (29.6 g, 90.85 mmol) contribute to anhydrous toluene (3 ml), and the resulting mixture is purged with argon and stirred for 16 hours at 120°C. After adding additional amounts of CuI (576 mg, 3.03 mmol), trans-(1R,2R)-N, N'-bismethyl-1,2-cyclohexanediamine (1.91 ml, 12.1 mmol) and Cs ₂ CO ₃ (20.0 g, 60.0 mmol) the reaction mixture was stirred for an additional 24 hours. The solvent was removed in vacuo, the residue was taken up in DCM and extracted with NaHCO ₃ solution (semi-conc.). The organic layer is dried over MgSO ₄ , filtered, the solvent is removed in vacuo and the residue is purified under NP-MPLC conditions. The product containing fractions (12) (HPLC-MS, method C: t _Ret. =1.62 min; MS (M+H) ⁺ =564) are combined and the solvent is removed in vacuo.

Step 9 4-((1-(2,6-difluoro-3-(propylsulfonamido)phenyl)-3-iodo-1H-pyrrolo[3,2-b]pyridin-5-yl)( methyl)amino)piperidine-1-carboxylic acid (13)

To a solution of sulfonamide (12) (1.078 g, 1.9 mmol) in DMF (4 ml)/THF (100 μl) was added N-iodosuccinimide (NIS) (474 mg, 2.1 mmol) and the mixture was stirred for 1 hours at room temperature. The reaction mixture was diluted with 30 ml DCM and extracted with NaHCO ₃ solution (semi-conc.). The combined organic layer is dried over MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography under RP HPLC conditions. The product containing fractions (13) (HPLC-MS, method B: t _Ret. =2.035 min; MS (M+H) ⁺ =688) was freeze dried.

Step 10 4-((1-(2,6-difluoro-3-(propylsulfonamido)phenyl)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine tert-butyl ester -5-yl)(methyl)amino)piperidine-1-carboxylic acid (15)

Sulfonamide (13) (770 mg, 1.12 mmol), pyrimidin-5-ylboronic acid (14) (194 mg, 1.57 mmol), Pd(dppf)Cl ₂ (82 mg, 0.11 mmol), LiCl (142 mg, 3.35 mmol) and Na ₂ CO ₃ (294 mg, 2.8 mmol) are placed in a mixture of dioxane/H ₂ O (2:1, 12 ml) and the resulting mixture is purged with argon and stirred for 1 hours at 100°C. The reaction mixture was diluted with DCM and extracted with a solution of NaHCO ₃ (semi concentrated). The organic layer was dried over MgSO ₄ , filtered, Isolute® was added, the solvent was removed in vacuo and the residue was purified under RP HPLC conditions. The product containing fractions (15) (HPLC-MS, method C: t _Ret. = 2.149 min; MS (M+H) ⁺ = 642) was freeze dried.

Step 11 N-(2,4-difluoro-3-(5-(methyl(piperidin-4-yl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine -5-yl)phenyl)propan-1-sulfonamide

To a solution of exemplary compound (15) (154 mg, 0.24 mmol) in DCM/MeOH (1:1, 4 mL) was added HCl (in dioxane, 4N, 2 mL) and the mixture was stirred for 3 hours at room temperature. The solvent is removed in vacuo. The resulting compound (16) (HPLC-MS, method B: t _Ret. =1.02 min; MS (M+H) ⁺ =542) was used without further purification.

Step 12 N-3-(5-((1-Ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1- yl)-2,4-difluorophenyl)propane-1-sulfonamide (I)

Compound I was prepared from compound (16) by reductive alkylation with acetaldehyde (40% in iPrOH) in the presence of 1.5 equivalents of sodium acetoxyborohydride in iPrOH. The crude product was recrystallized from ethanol to give the title compound in 84% yield.

Scaled synthesis of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1- yl)-2,4-difluorophenyl)propane-1-sulfonamide (BI 882370)

Step 1. N-(2,4-Difluoro-3-(5-(methyl(piperidin-4-yl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine -1-yl)phenyl)propan-1-sulfonamide

Isopropanol (8.83 kg) and compound (15) (1.80 kg, 2.8 mol) were added to the reactor, and the mixture was stirred and heated to 55~60°C. Concentrated hydrochloric acid (2.76 kg, 28 mol) was added dropwise to the reactor over 20 minutes at 60-65°C. Then the reaction mass was heated to 60-70°C and kept for 1 hour. The conversion was monitored by HPLC and reached about 99.5% after about 1 hour.

The reaction mass was cooled, and isopropanol was removed by distillation under reduced pressure at a temperature not exceeding 50°C. The resulting brown oil was dissolved in water (6.75 kg) and washed with extraction with ethyl acetate (2.02 kg) at 20-30°C. The aqueous phase was cooled to 15-20°C. The pH value was adjusted to 8.0-8.5 with 10% NaOH aqueous solution (~8.0 kg) at 20-30°C. The mixture was stirred for 3-4 hours at 20-30° C. with the pH adjusted to 8.0-8.5 by adding 10% NaOH aqueous solution every half an hour. The product was isolated by filtration and the filter cake was washed with water (3.6 kg). The solid was dried under vacuum at 45~50 until the water content reached no more than 5.5%. This yielded about 1.64 kg of crude compound (16) (yield 108% of theory; crude product formed containing water and NaCl). The raw product was used directly.

Step 12 N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1 -yl)-2,4-difluorophenyl) propane-1-sulfonamide (I)

Name Molecular mass Moley Weight Quantity Provider BI 878426 541.62 2.77 mol 1.50 kg 1.0 eq. Bioduro STAB (95%) 211.94 4.16 moles 0.93 kg 1.5 eq. dichloromethane No No 19.88 kg 10.0 volumes 40% acetal dehyde solution in DCM 44.05 9.71 moles 1.07 kg 3.5 eq. _H2O No No 30.0 kg 20.0 volumes Ammonia 17.03 16.61 moles 1.13 kg 6.0 eq. medical carbon No No 0.15 kg 10.0%

Methodology:

Dichloromethane (19.88 kg) and compound (16) (1.5 kg, 2.77 mol) were added to the reactor, and the mixture was stirred and cooled to 0-10°C under nitrogen atmosphere. Sodium triacetoxyborohydride (STAB) (95%, 0.93 kg, 4.16 mol) was added to the mixture at 0-10°C. The mixture was stirred for 20-30 minutes at 0-10°C. Acetaldehyde in DCM (40%, 1.07 kg, 9.71 mol) was slowly added to the mixture over 2 hours at 0-10°C. The reaction mixture was stirred at 0~10° C. under nitrogen atmosphere for 0.5-1 hour. The conversion rate was monitored by HPLC and reached about 99.5% after about 0.5-1 hour.

To the reaction mass was added water (15 kg) at a temperature below 15°C. The mixture was stirred at 15-30°C for 20~30 minutes. Aqueous ammonia (25%, 1.13 kg, 16.61 mol) was added to the mixture, and then the mixture was stirred for 0.5 hour. The organic phase was separated and then washed by extraction with water (15 kg) at 20-25°C. Activated carbon (0.15 kg) was added to the organic phase. The mixture was stirred for 1 hour and then filtered. The filtrate was concentrated under reduced pressure at a temperature not exceeding 40° C. to give compound (I) (1.58 kg, 100% yield) as a foamy solid.

Study of the crystallinity of the free base N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide

Study of the crystallinity of the free base N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl)propane-1-sulfonamide obtained by recrystallization from aqueous ethanol, which was used as a starting material for studying salt formation, showed that the compound had low crystallinity, as seen in FIG. 1.

Study of salt forms of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1 -yl)-2,4-difluorophenyl) propane-1-sulfonamide

Compound N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl )-2,4-difluorophenyl)propane-1-sulfonamide was mixed with different acids in different solvent systems.

A 96-well plate was dosed in MeOH (stock solution) at a concentration of approximately 40 mg/mL. This plate was placed in a vacuum oven to remove the liquid to obtain the same amount of solid material in each well. Various solvents/solvent mixtures and acids (approximately 500 µl) were then added to the solid material in each well, and the entire plate was heated to 50°C for 2 hours while stirring (using a small stirring arm placed in each hole).

The acids used were as shown in Table 1. The solvents used were as shown in Table 2. The crystallinity of the salts was obtained either in a slurry experiment or after crystallization by evaporation.

To study the formation of crystals in a suspension experiment, the obtained salts were studied by the XRPD method. An image of a multiwell plate showing the resulting salts is shown in FIG. 2A and XRPDs performed on the salt from each of the wells in the plate, showing the crystallinity of the salts formed, are presented in FIG. 2B.

To study the formation of crystals in the evaporation experiment, after a heating period, the solutions were filtered at the same temperature (50°C) using a preheated filter plate to ensure that undissolved material could not be transferred to other crystallization plates. The filtrate was dispensed into an evaporation plate (approximately 200 μl). The solvents were allowed to evaporate and the crystallinity of the resulting salts was examined by XRPD. An image of a multiwell plate showing the resulting salts is shown in FIG. 3A, and XRPD patterns performed on the salt from each of the wells in the evaporation plate, showing the crystallinity of the salts formed, are shown in FIG. 3B.

Table 1. Salts used to study the formation of salts

Experience Acid (please confirm the acids used) Code 1 Free base (no added acid) BS 2 Hydrochloric acid CL 3 Lemon acid CI 4 Wine acid TA 5 Fumaric acid FU 6 Salicylic acid SA 7 R-mandelic acid MY 8 Sulfuric acid SU 9 Phosphoric acid PH 10 succinic acid SC eleven Benzoic acid BZ 12 Maleic acid ML

Table 2. Solvents used to study the formation of salts

Experience Solvent A ethanol B Aqueous ethanol (90%) C 2-Propanol (iPrOH) D Aqueous iPrOH/(90%) E Acetone F ethyl acetate G Methyl Isobutyl Ketone (MIBK) H Tetrahydrofuran (THF)

Based on the initial selection, six crystalline salts formed, chloride (CL), fumarate (FU), phosphate (PH), succinate (SC), sulfate (SU) and tartrate (TA), as summarized in Table 3.

Table 3. Results of the initial study of salt forms

Salt form Number of manifestations Crystallinity Sample well (plate) Solvent for crystallization free base 0 amorphous Hydrochloride 9 High E2 Acetone Fumarate (Form I) 9 Medium F5 ethyl acetate Fumarate (Form II) 1 Medium D5 Aqueous iPrOH (90%) Tartrate 8 High F4 ethyl acetate Phosphate 8 Medium G9 MIBK Succinate 7 Medium F10 ethyl acetate Sulfate 8 Medium F8 ethyl acetate

Images performed on XRPD samples for each of the salt forms summarized in Table 3 are shown in FIG. 4.

Scaled synthesis of salt forms N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide

Based on the results of the initial identification of salt forms, chloride (CL), fumarate (FU), phosphate (PH), succinate (SC), sulfate (SU), and tartrate (TA) were resynthesized at a scale of about 50 mg. With the exception of phosphate, it was possible to obtain crystalline forms of each of chloride (CL), fumarate (FU), succinate (SC), sulfate (SU), and tartrate (TA), as summarized in Table 4.

Table 4. Results of the initial study of salt forms

Salt form Solvent for crystallization Crystallinity Notes Hydrochloride Acetone High Same as the hydrochloride from the original salt form study Tartrate ethyl acetate Medium Same as tartrate from original salt form research Fumarate (Form I) ethyl acetate medium high Same as Form I from the original study of the salt form Fumarate (Form II) iPrOH/H2O amorphous Sulfate ethyl acetate High Same as sulfate from the original salt form study Sulfate THF amorphous Phosphate MIBK amorphous Same as sulfate from the original salt form study Succinate ethyl acetate High ethyl acetate

Data for N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1 monohydrochloride -yl)-2,4-difluorophenyl)propane-1-sulfonamide obtained by scaling are shown in FIG. 5-9.

FIG. 5 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide with the sample from well E2 (lower graph) compared to the sample obtained from scaled synthesis (upper graph).

FIG. 6 is a DSC plot of the monohydrochloride salt of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide. The sample showed endotherm at about 321°C.

FIG. 7 is a TGA plot of the monohydrochloride salt of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide.

FIG. 8 is a sorption-desorption plot for N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide showing weight gain and loss when the relative humidity ranged from 0-100%. The sample showed a weight gain of approximately 2.4% in the range of 10-90% relative humidity.

FIG. 9 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide before (lower graph) and after (upper graph) the sorption-desorption experiment, showing that no significant change in the structure occurred.

Data for the monosuccinate salt N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl)propane-1-sulfonamide obtained by scaling are shown in FIG. 10-14.

FIG. 10 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide with the sample from well F10 (lower graph) compared to the sample obtained from scaled synthesis (upper graph).

FIG. 11 is a DSC plot of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide. The sample showed endotherm at about 182°C.

FIG. 12 is a TGA plot of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide.

FIG. 13 is a sorption-desorption plot for N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide showing weight gain and loss when the relative humidity ranged from 0-100%. The sample showed a weight gain of approximately 1.2% in the range of 10-90% relative humidity.

FIG. 14 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide before (lower graph) and after (upper graph) the sorption-desorption experiment, showing that no significant change in the structure occurred.

Synthesis and properties of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monohydrochloride salt -1-yl)-2,4-difluorophenyl) propane-1-sulfonamide

N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1- monohydrochloride salt yl)-2,4-difluorophenyl)propane-1-sulfonamide was obtained in various solvent systems by dissolving N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidine-5 -yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide (100 mg, 176 µmol) in various solvents (10-20 ml), by adding HCl (10 M solution in ethanol; 1 equivalent) at 50°C. The resulting solution was then heated with stirring at a temperature near the boiling point of the solvent for about 1 hour, then allowed to cool to room temperature and stirred for 3 hours or more (eg overnight). The salt was then recovered by filtration and dried at 50°C.

A summary of the experiments performed is presented in Table 5.

Table 5. Synthesis of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide

Solvent Salt output Purity by NMR Crystallinity Form EtOH (abs.) 79% >95% High A EtOH (90%) 71% >95% High A iPrOH (abs.) 76% >95% High A iPrOH (90%) 68% >95% High A Acetone 65% >95% High A ethyl acetate 73% >95% Medium A MIBK 74% >95% Medium A THF 76% >95% Low iPrOH (98%) 98% >95% High A ^# EtOH 95% High A ^#

# Synthesized in 1g scale

Preparative-scale synthesis of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide

N-(3-(5-((1-Ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl) -2,4-difluorophenyl)propane-1-sulfonamide (1.0 g, 1.76 mmol) in isopropanol (8 ml) was heated with stirring at 60° C. to form a brown solution. HCl (10 M solution in ethanol; 1.84 mmol, 1.05 equivalents) was added. The resulting suspension was heated with stirring at 60°C for 1 hour, then at 80°C for 1 hour, then at 50°C for 1 hour, and then left to cool to room temperature with stirring overnight (about 16 hours ). The salt was then separated by filtration and dried under reduced pressure at 50° C. overnight (about 16 hours). N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monohydrochloride salt was thereby obtained -1-yl)-2,4-difluorophenyl)propane-1-sulfonamide (1.023 g, 96% yield) as an off-white solid. Data for the monohydrochloride salt N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl)propane-1-sulfonamide are shown in FIG. 15-24.

FIG. 15 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide.

Table 6 lists representative XRPD peaks, d-values, and relative intensities for XRPD of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5- yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

Table 6. XRPD peaks, d-values and relative intensities for XRPD of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl) monohydrochloride salt -1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

Value 2θ (°) d-Value (Å) Relative intensity (I/I ₀ (%) 4.86 18.18 6 6.43 13.74 12 7.13 12.38 9 11.65 7.59 5 11.90 7.43 13 12.91 6.85 33 13.15 6.73 19 14.24 6.21 22 14.38 6.15 57 15.37 5.76 6 15.56 5.69 9 16.22 5.46 50 16.50 5.37 14 16.70 5.30 6 17.37 5.11 100 18.09 4.90 8 18.25 4.86 8 18.57 4.77 26 18.90 4.69 38 19.19 4.62 99 19.41 4.57 12 19.64 4.51 16 20.41 4.35 7 20.91 4.25 5 21.26 4.18 19 21.59 4.11 18 21.72 4.09 19 23.23 3.83 9 23.40 3.80 18 23.56 3.77 eleven 23.66 3.76 10 23.96 3.71 19 24.32 3.66 thirty 24.73 3.60 10 25.11 3.54 14 25.30 3.52 6 25.52 3.49 18 25.80 3.45 17 26.01 3.42 eleven 26.41 3.37 9 26.74 3.33 18 27.16 3.28 14 27.44 3.25 8 28.09 3.17 5 28.93 3.08 15 29.49 3.03 7 30.01 2.98 16 30.25 2.95 12 30.57 2.92 10 30.73 2.91 10 31.05 2.88 17 31.60 2.83 4 32.33 2.77 6 32.69 2.74 16 33.31 2.69 5 34.10 2.63 4 34.61 2.59 6 35.38 2.53 7 36.01 2.49 6 36.43 2.46 5 37.23 2.41 4 38.78 2.32 6 39.47 2.28 5 39.89 2.26 4

FIG. 16 is a DSC plot of the monohydrochloride salt of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide. The sample showed an endotherm with the beginning of the change at about 313°C. As a rule, the samples had an endotherm with the beginning of the change in the range of 312-322°C, corresponding to the melting point. Therefore, the melting point was measured at a value of about 317±5°C. Some samples had an additional endothermic event at about 250° C. corresponding to the loss of the solvent.

FIG. 17 is a TGA plot of the monohydrochloride salt of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide. The sample exhibited a loss on drying of about 1.7-2.5% up to about 250°C, corresponding to the release of water and solvent. The water content was measured at about 0.9% (Karl Fischer method).

FIG. 18 is a plot of the sorption-desorption isotherm for N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b ]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide showing weight gain and loss when the relative humidity ranged from 0-90%. The sample showed a reversible weight gain of approximately 4% in the range of 0-90% RH, and a reversible weight loss of approximately 2.5% in the range of 0-80% RH. The amount of absorbed and desorbed moisture appears as a function of the amount of organic solvent present in the sample.

FIG. 19 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide before (lower graph) and after (upper graph) the sorption-desorption experiment, showing that no significant change in the structure occurred.

FIG. 20 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide after grinding in a shaking mill for 10 minutes at 30 rpm. The sample showed a significant reduction in crystallinity, but no change in polymorph.

FIG. 21 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide after pressing at 2000 N for 1 second to form a tablet (5 mm diameter). The sample showed a slight decrease in crystallinity, but no change in polymorph.

FIG. 22 represents ¹ H-NMR spectrum (400 MHz, DMSO-d6) of the monohydrochloride salt of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl )-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 23 is a fragment of the ¹ H-NMR spectrum (400 MHz, DMSO-d6) in the aliphatic region (δ 0.5-6.0) of the monohydrochloride salt of N-(3-(5-((1-ethylpiperidin-4-yl) (methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 24 is a fragment of the ¹ H-NMR spectrum (400 MHz, DMSO-d6) in the aromatic region (δ 6.0-11.5) of the monohydrochloride salt of N-(3-(5-((1-ethylpiperidin-4-yl) (methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

The equilibrium solubility of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monohydrochloride salt was determined -1-yl)-2,4-difluorophenyl) propane-1-sulfonamide in various media. The results are summarized in Table 7.

Table 7 Solubility of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monohydrochloride salt pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide in various media

Wednesday Solubility after 2 hours (mg/ml) Water 0.1 HCl (0.1M) 0.06 HCl (0.01M) 0.02 McIlwain's buffer (pH 2.2) 0.40 McIlwain's buffer (pH 3.0) 0.20 McIlwain's buffer (pH 4.0) 0.10 McIlwain's buffer (pH 4.5) 0.20 McIlwain's buffer (pH 5.0) 0.20 Acetate buffer (pH 5.0) 0.20 McIlwain's buffer (pH 6.0) 0.10 McIlwain's buffer (pH 6.8) 0.10 Phosphate buffer (pH 6.8) 0.20 McIlwain's buffer (pH 7.4) 0.01 Sorensen's buffer (pH 9.0) 0.01 Sorensen buffer (pH 11) 0.70 0.1M NaOH >10.00 0.1 M citric acid 0.40 Model intestinal juice (after meals, FaSSIF) 0.002 Model intestinal juice (fasting, FeSSIF) 0.03

Intrinsic dissolution rate of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monohydrochloride salt -1-yl)-2,4-difluorophenyl)propane-1-sulfonamide in various media was measured in media with different pH values. The results are summarized in Table 8.

Table 8. Intrinsic dissolution rate of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2- b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide in various media at pH 1.0-7.4

medium pH Intrinsic dissolution rate (µg/ ^cm2 /min) 1.0 4 2.2 121 3.0 80 4.5 38 6.8 15 7.4 3

FIG. 25 is a graph showing the dissolution rate of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2] monohydrochloride salt -b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide over a period of 0-60 minutes in an aqueous medium at pH values from 1.0 to 7.4.

Synthesis and properties of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monosuccinate salt -1-yl)-2,4-difluorophenyl) propane-1-sulfonamide

Monosuccinate salt N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1- yl)-2,4-difluorophenyl)propane-1-sulfonamide was obtained in various solvent systems by dissolving N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidine-5 -yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide (100 mg, 176 mmol) in various solvents (about 8-10 ml per mg drug) when heated, and by adding succinic acid (1 equivalent). The resulting solution was heated with stirring at about 60-80°C for about 1 hour, then left to cool to room temperature and stirred for 3 hours or more (for example, overnight). Then the salt was isolated by filtration and dried at 50°C.

A summary of the experiments performed is given in Table 8.

Table 8. Synthesis of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide

Solvent Salt output Purity by NMR Crystallinity Form EtOH (abs.) 77% >95% High A iPrOH 85% >95% High A Acetone 85% >95% High F ^* ethyl acetate 84% >95% High A MIBK 0% (no crystallization) EtOH (abs.) 89% >95% High A EtOH (96%) 70% about 95% High A EtOH (96%) 75% >95% High A EtOH (96%) 75% >95% High A EtOH (98%) 92% >95% High A EtOH (abs.) 94% >95% High A EtOH (98%) 95% High A EtOH (97%) 92% >95% High A

* Acetone solvate

For the monosuccinate salt N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1 -yl)-2,4-difluorophenyl)propane-1-sulfonamide crystallization from a number of solvents gave the same crystal form (Form A) as obtained in the above experiments, but a different crystal form (Form B) (acetone solvate) obtained by crystallization from acetone.

Preparative-scale synthesis of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide (Forms A)

N-(3-(5-((1-Ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl) -2,4-difluorophenyl)propane-1-sulfonamide (800 mg, 1.40 mmol) in ethanol (8 ml) was heated with stirring at 60° C. to form a light yellow solution. Succinic acid (166 mg, 1.40 mmol, 1.0 equivalent) was added. The resulting solution was heated with stirring at 60° C. for 10 minutes, then at 45° C. for 10 minutes, until a precipitate formed with stirring. Then the suspension was heated at 55°C for 1 hour, and then left to cool to room temperature with stirring overnight (about 16 hours). Then the salt was separated by filtration and dried under reduced pressure at 50°C for 3 days. This gave N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monosuccinate salt -1-yl)-2,4-difluorophenyl)propane-1-sulfonamide (726 mg, 75% yield) as an off-white solid. Data for the resulting N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monosuccinate salt -1-yl)-2,4-difluorophenyl)propane-1-sulfonamide (Form A) are shown in FIG. 26-35.

FIG. 26 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide (Form A).

Table 9 lists representative XRPD peaks, d-values, and relative intensities for XRPD of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5- yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide (Form A).

Table 9. XRPD peaks, d-values and relative intensities for XRPD of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl) monosuccinate salt -1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide (Form A).

Value 2θ (°) d-Value (Å) Relative intensity (I/I ₀ (%) 5.63 15.68 20 6.72 13.15 5 7.26 12.16 1 10.05 8.80 13 11.28 7.84 32 12.15 7.28 20 12.72 6.95 17 12.94 6.84 17 13.45 6.58 3 13.85 6.39 25 14.20 6.23 3 14.56 6.08 38 14.87 5.95 9 15.13 5.85 6 15.44 5.73 100 15.86 5.58 38 16.13 5.49 63 16.97 5.22 12 17.15 5.17 40 17.33 5.11 4 17.54 5.05 6 17.85 4.97 13 18.00 4.92 37 18.67 4.75 4 19.14 4.63 47 19.48 4.55 2 19.82 4.48 43 20.01 4.43 79 20.20 4.39 62 20.54 4.32 60 21.01 4.23 10 21.48 4.13 44 21.78 4.08 74 22.68 3.92 10 23.07 3.85 6 24.17 3.68 23 24.38 3.65 13 24.60 3.62 16 24.86 3.58 15 25.37 3.51 5 25.67 3.47 17 25.79 3.45 17 26.05 3.42 21 26.23 3.39 20 26.39 3.37 12 26.61 3.35 3 27.06 3.29 21 27.42 3.25 eleven 27.77 3.21 eleven 27.90 3.19 8 28.25 3.16 7 28.64 3.11 24 29.45 3.03 9 29.79 3.00 3 30.02 2.97 5

FIG. 27 is a DSC plot of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide. The sample showed an endotherm with the beginning of the change at about 180°C. As a rule, the samples had an endotherm with the beginning of the change in the range of 179-182°C, corresponding to the melting point. Therefore, the melting point was measured at a value of about 180±3°C. Some samples had an additional endothermic event at about 160-165°C, corresponding to the loss of solvent.

FIG. 28 is a TGA plot of N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide. The sample exhibited a loss on drying of about 1.4-3.7% up to about 250°C, corresponding to the release of water and solvent. The water content was measured at about 0.03% (Karl Fischer method). Salt tends to include varying amounts of solvent in the crystals, which is released at temperatures higher than about 160°C.

FIG. 29 is a graph of the sorption-desorption isotherm for N-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b ]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide showing weight gain and loss when the relative humidity ranged from 0-90%. The sample showed a reversible weight gain of approximately 0.4% in the range of 0-90% RH, and a reversible weight loss of approximately 0.25% in the range of 0-80% RH. The amount of absorbed and desorbed moisture appears to depend on the amount of organic solvent present in the sample. Sorption and desorption were completely reversible with no change in crystallinity or polymorphic modification.

FIG. 30 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide before (lower graph) and after (upper graph) the sorption-desorption experiment, showing that no significant change in the structure occurred.

FIG. 31 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide after grinding in a shaking mill for 10 minutes at 30 rpm. The sample showed a slight decrease in crystallinity, but no change in polymorph.

FIG. 32 is an XRPD plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide after pressing at 2000 N for 1 second to form a tablet (5 mm diameter). The sample showed no significant reduction in crystallinity or change in polymorph.

FIG. 33 represents ¹ H-NMR spectrum (400 MHz, DMSO-d6) of the monosuccinate salt of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl )-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 34 is a fragment of the ¹ H-NMR spectrum (400 MHz, DMSO-d6) in the aliphatic region (δ 0.5-6.0) of the monosuccinate salt of N-(3-(5-((1-ethylpiperidin-4-yl) (methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

FIG. 35 is a fragment of the ¹ H-NMR spectrum (400 MHz, DMSO-d6) in the aromatic region (δ 6.0-10.0) of the monosuccinate salt of N-(3-(5-((1-ethylpiperidin-4-yl) (methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

The solubility of the monosuccinate salt N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl) propane-1-sulfonamide in various media. The results are summarized in Table 10.

Table 10 Solubility of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide in various media

Wednesday Solubility after 2 hours (mg/ml) Water >2.00 HCl (0.1 M) 0.20 HCl (0.01M) >0.05 McIlwain's buffer (pH 2.2) >5.00 McIlwain's buffer (pH 3.0) >5.00 McIlwain's buffer (pH 4.0) >5.00 McIlwain's buffer (pH 4.5) >5.00 McIlwain's buffer (pH 5.0) >5.00 Acetate buffer (pH 5.0) >5.00 McIlwain's buffer (pH 6.0) 3.7 McIlwain's buffer (pH 6.8) >5.00 Phosphate buffer (pH 6.8) >5.00 McIlwain's buffer (pH 7.4) 0.30 Sorensen's buffer (pH 9.0) 0.001 Sorensen buffer (pH 11) 0.05 0.1 M citric acid >5.00 Model intestinal juice (after meals, FaSSIF) 0.003 Model intestinal juice (fasting, FeSSIF) 0.40

Intrinsic dissolution rate of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monosuccinate salt -1-yl)-2,4-difluorophenyl)propane-1-sulfonamide in various media was measured in media with different pH values. The results are summarized in Table 11.

Table 11. Intrinsic dissolution rate of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2- b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide in various media at pH 1.0-7.4

medium pH Intrinsic dissolution rate (µg/ ^cm2 /min.) 1.0 (0.1 M HCl) 201 2.2 6844 3.0 6605 4.5 2457 6.8 2533 7.4 <1

FIG. 36 is a graph showing the dissolution rate of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2] monohydrochloride salt -b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide for a period of 0-60 minutes in an aqueous medium at pH values from 1.0 to 7.4.

Surprisingly, the solubility of the monosuccinate salt N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin- 1-yl)-2,4-difluorophenyl)propan-1-sulfonamide was found to be higher in some media tested than the solubility of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino) monohydrochloride salt )-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide. Intrinsic dissolution rate of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monosuccinate salt -1-yl)-2,4-difluorophenyl)propane-1-sulfonamide was also significantly higher than the intrinsic dissolution rate of the monohydrochloride salt N-(3-(5-((1-ethylpiperidin-4-yl)(methyl) amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

Pharmacokinetic parameters of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine- monohydrochloride salt 1-yl)-2,4-difluorophenyl)propane-1-sulfonamide and N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5- yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide in rat

Pharmacokinetic parameters of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine- monohydrochloride salt 1-yl)-2,4-difluorophenyl)propane-1-sulfonamide and N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5- yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide were measured at oral doses of 40 mg and 80 mg (doses calculated as free base) in Norwegian rats HsdHan: WIST Norway. Each salt was administered to 3 animals. The results are shown in Table 12.

Table 12 Mean plasma concentrations after oral administration of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[, monohydrochloride salt 3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide or N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino) monosuccinate salt -3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide to Norwegian rats (mean n=3±SD )

Time Plasma concentrations (nM) Monohydrochloride salt
(40 mg/kg) Monohydrochloride salt
(80 mg/kg) Monosuccinate salt (40 mg/kg) Monosuccinate salt
(80 mg/kg) 0.5 3.53 (±1.76) 523 (±303) 274 (±198) 1.0 6.56 (±5.7) 3.69 1320 (±691) 638 (±713) 2.0 107 (±124) 69.9 (±23.3) 6550 (±975) 3380 (±2720) 4.0 60.9 (±22.3) 419 (±32.6) 3860 (±671) 2620 (±1640) 6.0 13.3 (±8.45) 69 (±12.8) 1140 (±299) 1420 (±1300) 24.0 3.98 (±0.317) 5.09 (±198) PK parameters AUD range 0-6 hours 0-6 hours 0-24 hours 0-24 hours AUD (nM⋅h) 312 (±211) 1040 (±51.1) 30300 (±3120) 25100 (±20900) C _max (nM) 123 (±107) 419 (±32.6) 6550 (±975) 3520 (±2590) T _max (hour) 3.3 (±1.2) 4.0 (±0.0) 2.0 (±0.0) 2.7 (±1.2) Bioavailability after oral administration 1% 50%

The data show that the exposure values were significantly higher for the monosuccinate salt than for the monohydrochloride salt. In addition, oral bioavailability at 40 mg/kg was only 1% for the monohydrochloride salt, compared to 50% for the monosuccinate salt.

FIG. 37 is a plot of plasma concentration versus time values after oral administration to rats of (A) N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl) monohydrochloride salt )-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide at 40 mg/kg; (B) N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monohydrochloride salt -1-yl)-2,4-difluorophenyl) propane-1-sulfonamide at 80 mg/kg; (C) N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monosuccinate salt -1-yl)-2,4-difluorophenyl) propane-1-sulfonamide at 40 mg/kg; (D) N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monosuccinate salt -1-yl)-2,4-difluorophenyl) propane-1-sulfonamide at 80 mg/kg.

Scale-up preparation of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine- monosuccinate salt 1-yl)-2,4-difluorophenyl)propane-1-sulfonamide (Forms A)

Methanol (11.26 kg) and compound (I) (1.58 kg, 2.63 mol) were placed in the reactor, and the resulting mixture was stirred and heated to 60-70° C. until the solid dissolved. Succinic acid (393 g, 3.16 mol) was added to the mixture. Stirring at 60-70°C was continued for 0.5-1 hour, then the solution was cooled to 10~25°C. The mixture was seeded with Form A monosuccinate salt of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2 -b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide (3 g) and stirring was continued for an additional 1 hour until complete crystallization of the product. Then the mixture was cooled to 0-10°C and kept for 3-4 hours. The precipitated product was isolated by filtration and washed with chilled methanol. The solid was dried in vacuo at 45-50°C to form N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl) monosuccinate salt Form A -1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide (1.53 kg, 80.2% yield).

Additional characterization of salt forms of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide

Additional experiments were performed to characterize Form A of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2 -b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide. During characterization, some additional forms were identified.

Additional Common Methods

For the studies described below, unless otherwise noted, the instruments and methods described below were used.

X-ray powder diffractometry

XRPD analysis was performed using PANalytical Empyrean and X'Pert3 X-ray powder diffractometers, which were applied using the parameters listed in Table 13.

Table 13. XRPD parameters

Options Empyrean (reflect mode) X'Pert3 (reflect mode) x-ray radiation Cu, Kα, Kα1: 1.540598Å; Kα2: 1.544426Å; Kα2/Kα1 intensity ratio: 0.5 Cu, Kα, Kα1: 1.540598Å; Kα2: 1.544426Å; Kα2/Kα1 intensity ratio: 0.5 X-ray tube setup 45 kV, 40 mA 45 kV, 40 mA divergence gap automatically 1/8° Scan Mode continuous continuous Scan range (°2θ) 3°-40° 3°-40° Step range (°2θ) 0.0167° 0.0263° Scan step time (sec) 33.02 46.665 Test time (min.) ~10 minutes ~5 minutes

Thermogravimetric Analysis and Differential Scanning Calorimetry

TGA data were recorded using a TA Q500/Q5000/5500 TGA instrument from TA Instruments. DSC measurements were made using a TA Q200/Q2000/2500 DSC instrument from TA Instruments. The detailed parameters used are listed in Table 14.

Table 14. XPRD parameters

Options TGA DSC Method linear linear sample cuvette aluminum, open aluminum, twisted Temperature room-350°С 25-250°С Heating rate 10°C/minute 10°C/minute Purge gas nitrogen nitrogen

Dynamic vapor sorption

DVS measurements were carried out on an SMS (Surface Measurement Systems) DVS Intrinsic instrument. Relative humidity at 25° C. was calibrated against the melting point of LiCl, Mg(NO ₃ ) ₂ and KCl. The parameters for the DVS test are listed in Table 15.

Table 15. DVS parameters

Options DVS Temperature 25°C Sample size 10-200 mg Gas and flow rate nitrogen, 200 ml/minute dm/dt 0.002%/minute. Minimum duration of stability dm/dt 10 minutes Maximum equilibration time 180 minutes RH range 0% RH to 95% RH to 0% RH RH step size 10% RH 0% RH to 90% RH
5% RH 90% RH to 95% RH

Nuclear magnetic resonance

¹ H NMR measurements of solutions were performed using a Bruker 400 MHz NMR spectrometer using DMSO-d6 or deuterated methanol as solvent.

Microscopy with polarized light

PLM images were recorded using an Axio Lab upright microscope. A1 with ProgRes® CT3 camera at room temperature.

HLPC

HLPC analyzes were performed using an Agilent 1260 HPLC chromatograph. Chromatography conditions are listed in Tables 16 and 17.

Table 16. HLPC Parameters for Purity Test

HLPC Agilent 1260 with diode array (DAD) detector Column Gemini C18 110A, 250x4.6mm, 5µm mobile phase A: 0.1% TFA in H ₂ O; B: 0.1% in MeCN Gradient program Time (minutes) %IN 0.00 20 45.00 60 45.01 20 60.00 20 Time of processing 60.0 minutes Pause duration 0.0 minutes flow rate 1.0 ml/minute Injection volume 10 µl Detector wavelength UV at 234 nm Column temperature 30°C Sample temperature room temperature Diluent MeCN/H ₂ O (20:80 by volume)

Table 17. HLPC parameters for dissolution testing

HLPC Agilent 1260 with DAD Column Gemini C18 110A, 250x4.6mm, 5µm mobile phase A: 0.1% TFA in H ₂ O; B: 0.1% in MeCN Gradient program Time (minutes) % IN 0.00 20 5.00 70 5.10 20 10.00 20 Time of processing 10.0 minutes Pause duration 0.0 minutes flow rate 1.0 ml/minute Injection volume 10 µl Detector wavelength UV at 234 nm Column temperature 30°C Sample temperature room temperature Diluent MeCN

Polymorph selection experiments were carried out using antisolvent addition, antisolvent back addition, solid phase vapor diffusion, solution vapor diffusion, slurry, slow evaporation, slow cooling, and polymer-induced crystallization methods. In addition to Form A, four new forms of succinate were identified and designated as succinate Forms C, D, E, and F. All of these crystalline forms were characterized by X-ray powder diffractometry (XRPD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), proton nuclear magnetic resonance ( ¹ H-NMR) and HPLC. Two of the additional forms were anhydrous, one was a solvate, and one was not characterized due to limited sample quantities and reproducibility difficulties. All Forms C, D, E and F of the succinate salt were inferior to those of Form A in terms of stability and other characteristics.

Further characterization of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt Form A pyridin-1-yl)-2,4-difluorophenyl) propane-1-sulfonamide

A sample of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2- b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

XRPD results of two batches of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2- b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide are shown in FIG. 38.

The results of TGA and DSC analysis are shown in FIG. 39. The TGA curve of succinate Form A showed a weight loss of 1.9% to 140°C. On the DSC curve, the step of the endormic signal at 159.5°C (the beginning of the change) was observed before the sharp endotherm at 181.3°C (the beginning of the change).

The ¹ H-NMR spectra are shown in FIG. 40 (with DMSO-d6 as solvent) and FIG. 41 (with deuterated MeOH as solvent), and the molar ratio of succinic acid to free base was determined to be 0.9:1. The acid/free base molar ratio in Type A succinate was calculated using peak integration at 2.38 ppm. (integrated intensity 3.58, succinate hydrogen atoms, 4H) and at 1.78 ppm. (integral intensity 6.00, free base hydrogen atoms, 6H).

HPLC analysis confirmed that the purity of the succinate Form A sample (810023-01-A) was 99.5% (total peak area).

Additional XRPD, TGA, DSC and ¹ H-NMR analyzes were performed after heating the succinate Form A to 170°C and then cooling to room temperature. The results were shown in FIG. 42-45.

It was confirmed by XRPD (FIG. 42) that after heating, the crystalline form was still succinate Form A.

In the TGA curve (FIG. 43), a weight loss of 2.6% up to 140° C. was observed, which was likely due to moisture adsorbed to the surface when the heated sample was exposed to air prior to additional TGA testing. TGA analysis was performed approximately 4.5 hours after the heating experiment. The weight loss of the heated sample (2.6%) was higher than that observed initially (1.9%), indicating that the hygroscopicity of the succinate Form A may increase slightly after heat treatment.

On the DSC curve (FIG. 44) was observed only one endotherm at 182.7°C (the beginning of the change).

In NMR analysis (FIG. 45), the molar ratio of succinic acid/free base was still observed to be 0.9:1.

Based on the above results, Type A succinate was identified as anhydrous.

Equilibrium solubility

The equilibrium solubility of Form A in water was measured at room temperature. The solid was suspended in H ₂ O (with a stirring speed of ~800 rpm) at room temperature. After 24 hours, the suspension was centrifuged (10000 rpm, 5 minutes) followed by filtration (through a PTFE membrane with a pore size of 0.45 μm). The supernatant (the first few drops were discarded) was analyzed by HPLC and the pH was measured. The remaining solid was analyzed by XRPD. The measured solubility of Form A was 14.3 mg/ml, respectively.

XRPD analysis showed no shape change for Form A (FIG. 46) during the dissolution experiment.

Stability in the solid state

Samples of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridine monosuccinate salt Form A -1-yl)-2,4-difluorophenyl)propane-1-sulfonamide was kept for storage at 80°C for one day, at 25°C under conditions of 60% RH and at 40°C under conditions of 75% - noah RH for one week. All samples were then characterized using XRPD and HPLC, with the results summarized in Table 18.

No shape change or decrease in HPLC purity was observed for Form A under any of the conditions, indicating that Form A of the N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3 -(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propan-1-sulfonamide has good solid state stability. The results of the XRPD analysis are shown in FIG. 47.

Table 18 Summary of Solid State Stability Evaluation Results of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H- Monosuccinate Salt Form A pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide

Conditions: 80%C/1 day 25°C, 60% RH/1 week 25°C, 60% RH/1 week Initial Purity: Purity (% of initial) Shape change Purity (% of initial) Shape change Purity (% of initial) Shape change 99.5% 99.9 No 100.0 No 100.0 No

Hygroscopicity

To investigate the stability of the solid form as a function of moisture, the DVS isotherm of Form A of the monosuccinate salt of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl )-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide at 25° C. between 0 and 95% RH.

DVS plot of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b] monosuccinate salt Form A pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide shown in FIG. 48 shows 0.14% water uptake at 25°C/80% RH, confirming that Form A is not hygroscopic.

XRPD analysis, as shown in FIG. 49, showed that DVS analysis did not result in any form change for N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino) monosuccinate salt Form A )-3-(pyrimidin-5-yl)-1H-pyrrolo[3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide.

Microscopy with polarized light

PLM images were taken to study the morphology of N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[] monosuccinate salt Form A 3,2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide. As shown in FIG. 50, Form A consists of rod-shaped crystals.

Conclusion

Based on all of the above results, N-(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1H-pyrrolo[3, 2-b]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide was found to be a stable, non-hygroscopic crystalline form that was suitable for pharmaceutical use.

Various modifications of the invention, in addition to those described here, will be apparent to those skilled in the art from the above description. Such modifications are also intended to be within the scope of the appended claims. Each reference, including all patents, patent applications and publications cited in this application, is incorporated into this description by reference in its entirety.

Claims (7)

1. The method of obtaining a crystalline monosuccinate salt N - (3- (5- ((1-ethylpiperidin-4-yl) (methyl) amino) -3- (pyrimidin-5-yl) -1 H -pyrrolo[3,2- b ]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide having an x-ray powder diffraction pattern containing at least five of the following peaks, in 2θ units: 15.4° ± 0.5°; 16.1° ± 0.5°; 17.2° ± 0.5°; 19.1° ± 0.5°; 19.8° ± 0.5°; 20.0° ± 0.5°; 20.2° ± 0.5°; 20.5° ± 0.5°; 21.5° ± 0.5°; and 21.8 ° ± 0.5 °, including the interaction of N - (3- (5- ((1-ethylpiperidin-4-yl) (methyl) amino) -3- (pyrimidin-5-yl) -1 H - pyrrolo[3,2- b ]pyridin-1-yl)-2,4-difluorophenyl)propane-1-sulfonamide with one equivalent of succinic acid in a solvent, where the solvent is selected from C _1-4 alcohol or aqueous C _1-4 -alcohol, or from ethyl acetate, where the method includes: heating and stirring N -(3-(5-((1-ethylpiperidin-4-yl)(methyl)amino)-3-(pyrimidin-5-yl)-1 H -pyrrolo[3,2- b ]pyridine- 1-yl)-2,4-difluorophenyl)propane-1-sulfonamide with one equivalent of succinic acid in a solvent at 30-100°C for 5 minutes to 4 hours to form a suspension, cooling and stirring the suspension to 0-25°C for 5 minutes to 24 hours, filtering the crystalline salt from the slurry, and drying the crystalline salt under reduced pressure at 50° C. to obtain a crystalline salt. 2. The method of claim 1 further comprising crystallizing the salt from ethanol, isopropanol, aqueous ethanol or aqueous isopropanol, or from ethyl acetate.

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