Sulfasalazine salts, production processes, and uses

Abstract

To provide a process for preparing crystalline organic salt of sulfasalazine, and a pharmaceutical composition comprising a therapeutically effective amount of one or more of the crystal salt forms of sulfasalazine.SOLUTION: There is provided a process for preparing crystalline organic salt of sulfasalazine, wherein the organic salts are selected from diethylamine, and piperazine. The process comprises the following steps: A1: providing sulfasalazine free acid form in suitable solvents, wherein the solvents are selected from the group consisting of acetone, acetonitrile, and alcohol such as methanol, ethanol, and propanol; B1: providing an organic amine-containing constituent selected from piperazines in suitable solvents, wherein the solvents are or comprise water; C1: mixing sulfasalazine solution of step A1) with the organic amine containing constituent solution of step B1) at room temperature; and D1: separating the crystals of piperazinesulfasalazine Form A or a solvate thereof formed in the solutions of step C1).SELECTED DRAWING: Figure 3

Description

[Technical field]

[0001] The present invention relates to novel crystalline salt forms of sulfasalazine obtainable by using the preparation methods of the present invention, in particular crystalline Form A of the D(-)-N-methylglucamine (meglumine) salt of sulfasalazine (see Figure 1), crystalline Form A of the piperazine salt of sulfasalazine (see Figure 2) and crystalline Form B of the diethylamine salt of sulfasalazine (see Figure 4), as well as pharmaceutical compositions comprising one or more of the crystalline salt forms of the present invention for the treatment of diseases or conditions in which modulation of inflammatory cells is beneficial, in particular diseases or conditions involving bone or joints and / or the gastrointestinal tract. [Background technology]

[0002] The compound known by the generic name sulfasalazine (also known as (3Z)-6-oxo-3-[[4-(pyridin-2-ylsulfamoyl)phenyl]hydrazinylidene]cyclohexa-1,4-diene-1-carboxylic acid (IUPAC); 2-hydroxy-5-[2-[4-[(2-pyridinylamino)sulfonyl]phenyl]diazenyl]-benzoic acid (CA index name)) was first disclosed in U.S. Pat. No. 2,396,145 (GB 564990) and is highly effective in treating various autoimmune diseases, such as rheumatoid arthritis, juvenile idiopathic arthritis, ankylosing spondylitis, ulcerative colitis and Crohn's disease.

[0003] Sulfasalazine forms tan crystals (molecular weight 398.39 g / mol). The melting point has been determined to be 240°C-245°C (US 2,396,145). The solubility of sulfasalazine in water is less than 5 mg / 100 ml. The material has four theoretical pk values ​​of 0.6, 2.4, 9.7, and 11.8. a / b It has proven extremely difficult to prepare hydrate- and solvate-free salts of sulfasalazine using established methods.

[0004] The structure of sulfasalazine ((3Z)-6-oxo-3-[[4-(pyridin-2-ylsulfamoyl)phenyl]hydrazinylidene]cyclohexa-1,4-diene-1-carboxylic acid (IUPAC); 2-hydroxy-5-[2-[4-[(2-pyridinylamino)sulfonyl]phenyl]diazenyl]-benzoic acid (CA Index Name)) is shown below: [ka]

[0005] Sulfasalazine is a well-established active pharmaceutical ingredient used in anti-inflammatory therapy. It is used in the treatment of active rheumatoid arthritis, active juvenile idiopathic oligoarthritis, active juvenile idiopathic polyarthritis and spondyloarthropathy with peripheral arthritis in humans. Sulfasalazine is also used as a prodrug of 5-aminosalicylic acid in the treatment of inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis. In adults, sulfasalazine is generally administered orally in tablets at doses of 500 to 4000 mg / day, based on the tolerability of sulfasalazine and its efficacy.

[0006] Sulfasalazine is one of the most widely used disease-modifying antirheumatic drugs (DMARDs) and is also used in combination with glucocorticoids and / or in combination with other small molecule DMARDs, such as methotrexate and / or hydroxychloroquine and / or biologic DMARDs, such as TNF-α related biologics.

[0007] The mechanism of action of sulfasalazine and its metabolites 5-aminosalicylic acid and sulfapyridine is still partially unknown. Sulfasalazine and / or its metabolites have anti-inflammatory and immunomodulatory properties in vivo and in vitro in various (inflammatory) cell types, such as T cells, dendritic cells, macrophages, natural killer cells, epithelial cells, B cells and mast cells, through different biological "pathways". For example, it has been shown that sulfasalazine-treated dendritic cells are unable to stimulate T cells due to inhibition of the NF-kB pathway (Matasic R, Dietz AB, Vuk-Pavlovic S.; "Maturation of human dendritic cells as sulfasalazine target."; Croat Med J 2001 Aug; 42(4): 440-5). Sulfasalazine has also been shown to inhibit the binding of TNF-α to its receptor in 125I-TNF-α displacement studies. In addition, sulfasalazine, like methotrexate, has been shown to increase adenosine release by inhibiting AICAR transformylase, thereby reducing inflammation (P Gadangi, M Longaker, D Naime, RI Levin, PA Recht, MC Montesinos, MT Buckley, G Carlin and BN Cronstein; "The anti-inflammatory mechanism of sulfasalazine is related to adenosine release at inflamed sites.", J Immunol March 1, 1996, 156 (5) 1937-1941).The well-established antioxidant effect of sulfasalazine, in conjunction with its inhibitory effect on neutrophil oxidative burst, has been shown to be exerted both by its scavenging effect on reactive oxygen and nitrogen species as well as by its metal chelating properties (Couto D1, Ribeiro D, Freitas M, Gomes A, Lima JL, Fernandes E., Redox Rep. 2010;15(6):259-67. doi: 10.1179 / 135100010X12826446921707, "Scavenging of reactive oxygen and nitrogen species by the prodrug sulfasalazine and its metabolites 5-aminosalicylic acid and sulfapyridine.").

[0008] However, due to the low solubility of sulfasalazine (0.03 mg / mL in deionized water at 24 °C) and the currently known pharmaceutical compositions of sulfasalazine, the systemic bioavailability of sulfasalazine in humans is low (approximately 15-20% of an oral dose is absorbed in the small intestine) and the intra- and inter-intestinal variability of pharmacokinetics is high (C max (The median time to absorption is 4-12 hours, with a median peak concentration of 6 hours.) Unabsorbed sulfasalazine is converted by the aza-reducing gut flora to 5-aminosalicylic acid (10-30% of systemic bioavailability) and sulfapyridine (approximately 60% of systemic bioavailability). Metabolites can be detected in plasma after approximately 10 hours. The half-life of intravenously administered sulfasalazine is approximately 7.6 ± 3.6 hours.

[0009] Apart from the biotransformation of sulfasalazine by the gut flora in the lower gastrointestinal tract, sulfasalazine is also metabolized in the liver to the metabolites 5-aminosalicylic acid and sulfapyridine. In the liver, the primary metabolite sulfapyridine is acetylated before excretion, the rate of which is determined by the acetylation phenotype. Thus, the half-life of sulfapyridine can vary from 10.4 to 14.8 hours (depending on the fast or rather slow acetylation phenotype).

[0010] The most common side effects associated with sulfasalazine are anorexia, headache, nausea, vomiting, stomach discomfort, although apparent reversible oligospermia, dizziness, fever, asthenia, insomnia and vertigo may also affect patients while taking sulfasalazine.

[0011] However, gastrointestinal reactions including hepatitis, liver failure, pancreatitis, bloody diarrhea, folate malabsorption, digoxin malabsorption, stomatitis, diarrhea, abdominal pain, and neutropenic enterocolitis may also occur during administration.

[0012] Also, the skin (e.g. skin rash or pruritus, hives, increased sensitivity to sunlight), blood / lymphatic system (aplastic anemia, agranulocytosis, leukopenia, megaloblastic (macrocytic) anemia, purpura, thrombocytopenia, thrombocytopenia, methemoglobinemia, congenital neutropenia and myelodysplastic syndromes) or central nervous system (transverse myelitis, convulsions, meningitis, posterior spinal transient lesions, cauda equina syndrome, Guillain-Barre syndrome, peripheral neuropathy, mental depression, vertigo, hearing loss, insomnia, ataxia, hallucinations, tinnitus and drowsiness). The hepatobiliary damage that occurs can be hepatotoxicity including elevated liver function tests (SGOT / AST, SGPT / ALT, GGT, LDH, alkaline phosphatase, bilirubin), jaundice, cholestatic jaundice, cirrhosis, cholestatic hepatitis, cholestasis and potentially hepatocellular injury including hepatic necrosis and liver failure. Some of these cases have been fatal.

[0013] The low absorption rate of sulfasalazine is one reason why large amounts of sulfasalazine, up to 500 mg per single solid oral dosage form, need to be administered to patients to obtain sufficient exposure to the drug sulfasalazine and thus sufficient clinical effect. However, large amounts of drug substance and / or large tablet size can result in poor patient compliance and unnecessarily high rates of adverse events, in part due to undesirable ratios of sulfasalazine and its metabolites in vivo.

[0014] In the manufacture of pharmaceutical compositions, it is important that the active compound is in a form that can be conveniently handled and processed to obtain a commercially viable manufacturing process.In this regard, the chemical and physical stability of the active compound are important factors.The active compound and the compositions containing it must be able to be effectively stored for a significant period of time without showing significant changes in physicochemical properties (e.g. chemical composition, density, moisture content and solubility).

[0015] Therefore, it would be beneficial to provide a solvate-free crystalline form of sulfasalazine that exhibits solubility.From the prior art, it is evident that it is extremely difficult to prepare and / or isolate solvate-free crystals of sulfasalazine while at the same time improving solubility.

[0016] It is known that sulfasalazine has metal chelating properties in vivo. However, monosalts of sulfasalazine with counterions sodium and potassium have been mentioned in Nygard et al. (Nygard, B.; Olofsson, J. and Sandberg, M.: "Some physicochemical properties of salicylazosulphapyridine, including its solubility, protolytic constants and general spectrochemical and Polarographic behavior.", Acta Pharmaceutica Suecica 3: 313-342 (1966)) but have not been isolated. Hemisalts of sulfasalazine with metal counterions such as strontium (as trihydrate), calcium (as trihydrate), and magnesium (as trihydrate) are less soluble than sulfasalazine. Other metal complexes of sulfasalazine with ammonium (cerium, thorium and uranium) have been tested as well (GG Mohamed et al. Spectrochim Acta A Mol Biomol Spectrosc 62 (4-5), 1095-1101. 2005 Jun 13; "Structural and Thermal Characterization of Cerium, Thorium and Uranyl Complexes of Sulfasalazine"). However, these counterions are not pharmacologic accepted.

[0017] Conformational analysis of sulfasalazine salts with various counterions such as Mg, Sr, Ca and Zn indicates that the terminal pyridine ring exhibits a degree of oriental flexibility, which indicates a propensity for conformational polymorphism of sulfasalazine salts.

[0018] Patent application GB 1,166,684 discloses alkoxy-amine addition salts of sulfasalazine. According to the patent, the prepared alkoxy-amine addition salts of sulfasalazine are difficult to crystallize in that they are obtained as viscous oils that crystallize only when stirred with ether or alcohol, and many of the disclosed salts are highly hygroscopic and / or have high water content. For example, N-methyl-(1)-D-glucosamine salt of sulfasalazine (also known as meglumine salt, as used below) is identified in GB 1,166,684 and is prepared by adding a solution of methylglucamine in hot methyl glycol to a solution of sulfasalazine in 2-methoxyethanol, and shows a water content of 9% after thorough drying. Such a water content is particularly unsuitable for the preparation of a stable pharmaceutical composition.

[0019] Due to their unfavourable physicochemical properties (hygroscopicity and / or high water content and / or low solubility and / or pharmaceutical unacceptability), the sulfasalazine salts disclosed thus far are not considered suitable for use in pharmaceutical compositions.

[0020] CN 106 279 008 A relates to the technical field of a method for purifying sulfasalazine (5-[p-(2-pyridylaminosulfonyl)benzene]azo-salicylic acid). Sulfasalazine is purified by using a specific amine salt of sulfasalazine as an intermediate product and precipitating sulfasalazine from a solution containing the specific amine salt. According to Example 6, a specific diethylamine salt of sulfasalazine is prepared as an intermediate product, and then sulfasalazine is precipitated from the solution. The applicant has prepared a sulfasalazine diethylamine salt intermediate according to the description provided in Example 6 of CN 106 279 008 A. The diethylamine salt of sulfasalazine was also evaluated by X-ray powder diffraction (XRPD), which demonstrated that the obtained diethylamine sulfasalazine exists as Form A crystals of the diethylamine salt of sulfasalazine according to FIG. 3.

[0021] Thus, there remains a need to provide pharmaceutical compositions of sulfasalazine that increase the bioavailability of sulfasalazine and / or the solubility of sulfasalazine, and / or improve the risk-benefit ratio of pharmaceutical sulfasalazine compositions, particularly to reduce the severity of adverse events and / or improve patient compliance. Summary of the Invention

[0022] The object of the present invention is achieved by the subject matter of the independent claims. Advantages (preferred embodiments) are shown in the following detailed description including the drawings and in the dependent claims.

[0023] Accordingly, a first aspect of the present invention is a method for preparing crystals of an organic salt of 2-hydroxy-5-[2-[4-[(2-pyridinylamino)sulfonyl]phenyl]diazenyl]-benzoic acid (sulfasalazine), the organic salt being selected from diethylamine, piperazine and D-(−)-N-methylglucamine (meglumine), the method comprising the steps of: A1: providing sulfasalazine free acid form in a suitable solvent; B1: Providing an organic amine-containing component selected from meglumine and piperazine in a suitable solvent; C1: mixing the sulfasalazine solution of step A1) with the organic amine-containing component solution of step B1) at room temperature, preferably 19°C to 25°C; and D1: isolating the crystals of piperazine sulfasalazine Form A or a solvate thereof or the crystals of meglumine sulfasalazine Form A or a solvate thereof formed in the solution of step C1); or A2: Providing sulfasalazine free acid form in a suitable solvent; B2: Providing an organic amine-containing component selected from meglumine and diethylamine in a suitable solvent; C2: mixing the sulfasalazine solution of step A2) with the organic amine-containing component solution of step B2), wherein the amine-containing component is in at least a 5% molar excess relative to the sulfasalazine, more preferably at least a 10% molar excess, more preferably at least a 20% molar excess; and D2a: Concentrating the mixture formed in step C2) to separate crystals of meglumine sulfasalazine Form A or a solvate thereof or crystals of diethylamine sulfasalazine Form B or a solvate thereof; or D2b: adding an additional solvent to the mixture formed in step C2), wherein the additional solvent is different from the solvent used in steps A2) and B2), and isolating the crystals of meglumine sulfasalazine Form A or a solvate thereof or the crystals of diethylamine sulfasalazine Form B or a solvate thereof; The present invention relates to a process for the preparation of crystalline Form A of diethylamine sulfasalazine, comprising or consisting of:

[0024] A second aspect of the present invention relates to crystalline meglumine sulfasalazine Form A, crystalline piperazine sulfasalazine Form A or crystalline diethylamine sulfasalazine Form B of the present invention, each obtained according to the process of the present invention.

[0025] A third aspect of the invention relates to pharmaceutical compositions comprising a therapeutically effective amount of one or more of the crystalline salt forms of sulfasalazine of the present invention.

[0026] A fourth aspect of the present invention is i) a human disease or condition in which modulation of inflammatory cells is beneficial; ii) diseases or conditions relating to bone or joints, preferably arthritis associated with or including osteoarthritis, primary or secondary to, for example, congenital hip dysplasia; cervical and lumbar spondylitis, and lower back and neck pain; rheumatoid arthritis and Still's disease; seronegative spondyloarthropathy, such as ankylosing spondylitis, psoriatic arthritis, reactive arthritis and undifferentiated spondarthropathy, septic arthritis and other arthopathies and bone disorders associated with infections, such as tuberculosis, e.g. Pott's disease and Ponce's disease; acute and chronic crystal-induced synovitis, such as urate gout, calcium pyrophosphate deposition disease, and calcium apatite associated tendonitis. tendon), bursal and synovial inflammation; Behcet's disease; primary or secondary Sjogren's syndrome; systemic sclerosis and localized scleroderma; systemic lupus erythematosus, mixed connective tissue disease, and undifferentiated connective tissue disease; inflammatory myopathies, such as dermatomyositis and polymyositis; polymyalgia rheumatica; juvenile arthritis, such as idiopathic inflammatory arthritis of any joint location and associated syndrome, and rheumatic fever and its systemic complications; vasculitis, such as giant cell arteritis, Takayasu's arteritis, Churg-Strauss syndrome, polyarteritis nodosa, microscopic polyarteritis, and vasculitis associated with viral infections, hypersensitivity reactions, cryoglobulins, and paraproteins; lower back pain; Familial Mediterranean Fever, Muckle-Wells syndrome, and Familial Hibernian Fever, Kikuchi's disease; a disease or condition selected from the group consisting of drug-induced joint pain, tendonitis, and myopathy; and iii) diseases or conditions relating to the digestive tract, preferably selected from the group consisting of eosinophilic gastroenteritis, mastocytosis, Crohn's disease, colitis, e.g. ulcerative colitis, proctitis; celiac disease, irritable bowel syndrome, and food-related allergies that may affect areas distant from the intestine, e.g. migraine, rhinitis or eczema. The present invention relates to the use of a crystalline salt form of sulfasalazine according to the present invention or a pharmaceutical composition according to the present invention in the manufacture of a medicament for / in the treatment of

[0027] A fifth aspect of the present invention is i) a human disease or condition in which modulation of inflammatory cells is beneficial; ii) diseases or conditions relating to bone or joints, preferably arthritis associated with or including osteoarthritis, primary or secondary to, for example, congenital hip dysplasia; cervical and lumbar spondylitis, and lower back and neck pain; rheumatoid arthritis and Still's disease; seronegative spondyloarthropathy, such as ankylosing spondylitis, psoriatic arthritis, reactive arthritis and undifferentiated spondarthropathy, septic arthritis and other arthopathies and bone disorders associated with infection, such as tuberculosis, e.g. Pott's disease and Ponce's disease; acute and chronic crystal-induced synovitis, such as uric acid gout, calcium pyrophosphate deposition disease, and calcium apatite-related tendon, bursa and synovial inflammation; Behcet's disease; primary or secondary Sjogren's syndrome. systemic sclerosis and localized scleroderma; systemic lupus erythematosus, mixed connective tissue disease, and undifferentiated connective tissue disease; inflammatory myopathies, e.g., dermatomyositis and polymyositis; polymyalgia rheumatica; juvenile arthritis, e.g., idiopathic inflammatory arthritis of any joint location and associated syndrome, and rheumatic fever and its systemic complications; vasculitis, e.g., giant cell arteritis, Takayasu's arteritis, Churg-Strauss syndrome, polyarteritis nodosa, microscopic polyarteritis, and vasculitis associated with viral infections, hypersensitivity reactions, cryoglobulins, and paraproteins; lower back pain; Familial Mediterranean Fever, Muckle-Wells syndrome, and Familial Hibernian Fever, Kikuchi's disease; drug-induced arthralgia, tendonitis, and myopathy; and iii) diseases or conditions relating to the digestive tract, preferably selected from the group consisting of eosinophilic gastroenteritis, mastocytosis, Crohn's disease, colitis, e.g. ulcerative colitis, proctitis; celiac disease, irritable bowel syndrome, and food-related allergies that may affect areas distant from the intestine, e.g. migraine, rhinitis or eczema. The present invention relates to a method for treating a disease or condition in a patient suffering from or at risk of having said disease or condition, comprising administering to the patient a therapeutically effective amount of a crystalline salt form of sulfasalazine of the present invention or a pharmaceutical composition of the present invention.

[0028] The aspects of the invention described below may also include any possible combination of the preferred embodiments described in the dependent claims or disclosed in the following detailed description and drawings, if reasonable to a person skilled in the art. [Brief description of the drawings]

[0029] [Figure 1] 1 shows a powder X-ray diffraction pattern of crystalline Form A of the meglumine salt of sulfasalazine. [Diagram 2] 1 shows a powder X-ray diffraction pattern of crystalline Form A of the piperazine salt of sulfasalazine. [Diagram 3] 1 shows a powder X-ray diffraction pattern of crystalline Form A of the diethylamine salt of sulfasalazine. [Figure 4] 1 shows a powder X-ray diffraction pattern of crystalline Form B of the diethylamine salt of sulfasalazine. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0030] The present inventors have discovered that novel and inventive crystalline salt forms of sulfasalazine, in particular crystalline Form A of the D(-)-N-methylglucamine (meglumine) salt of sulfasalazine (see FIG. 1), crystalline Form A of the piperazine salt of sulfasalazine (see FIG. 2) and crystalline Form B of the diethylamine salt of sulfasalazine (see FIG. 4), exhibit a favorable pharmacokinetic profile as compared to the free acid form of sulfasalazine: max Decrease in C max and increased F (bioavailability) (see Example 7 below).

[0031] The inventors of the present application have also demonstrated that the free acid form of sulfasalazine has low gastrointestinal absorption (see Arik Dahan, Gordon L. Amidon; "Small intestinal efflux mediated by MRP2 and BCRP shifts sulfasalazine intestinal permeability from high to low, enabling its colonic targeting"; American Journal of Physiology - Gastrointestinal and Liver Physiology Published 21 July 2009 Vol. 297 no. 2, G371-G377) and low permeability through Caco-2 cell monolayers (Liang E1, Chessic K, Yazdanian M.; "Evaluation of an accelerated Caco-2 cell permeability model."; J Pharm Sci. 2000 Mar;89(3):336-45) compounds, the salt crystalline forms of sulfasalazine of the present invention have instead been found to exhibit improved permeability through Caco-2 cell monolayers as compared to the free acid form of sulfasalazine. According to alternative or additional embodiments of all aspects of the present invention, the apparent apical to basolateral permeability coefficient of the crystalline forms of sulfasalazine of the present invention, preferably Form A crystals of the meglumine salt of sulfasalazine, is increased by ≧0.5, ≧1.0, ≧1.5, ≧2.0, ≧2.5, ≧3.0 or ≧3.5 fold as compared to the free acid form of sulfasalazine (see Example 6 below).

[0032] Additionally, the free acid form of sulfasalazine is classified as practically insoluble (<0.1 mg / mL) according to the United States Pharmacopeia, with a solubility of 0.031 mg / mL in deionized water at 24° C. According to measurements performed by the inventors, the free acid form of sulfasalazine exhibits a solubility of 0.06 mg / mL in deionized water at 24° C., whereas the salt crystalline forms of sulfasalazine of the present invention typically exhibit an increased solubility of ≧0.1 mg / mL, more preferably ≧0.5 mg / mL, ≧1 mg / mL, ≧5 mg / mL, ≧10 mg / mL, ≧15 mg / mL, ≧20 mg / mL, ≧25 mg / mL, ≧30 mg / mL or ≧35 mg / mL in deionized water at 24° C. By separate measurements, the crystalline forms of sulfasalazine of the present invention, particularly crystalline Form A of the meglumine salt of sulfasalazine, exhibit solubilities of ≧50 mg / mL, ≧60 mg / mL, ≧70 mg / mL, ≧80 mg / mL, ≧90 mg / mL, ≧100 mg / mL in deionized water at 22° C. and pH 6.6 (see Example 8 below).

[0033] Thus, due to increased bioavailability and / or solubility, reduced doses of sulfasalazine may be used in the therapeutic treatment of diseases or conditions where modulation of inflammatory cells is beneficial, such as rheumatoid arthritis, ankylosing spondylitis, and juvenile idiopathic arthritis, where systemic exposure to sulfasalazine is necessary, without altering total exposure to sulfasalazine. This results in an improved risk-benefit profile, as it reduces exposure to sulfapyridine, a metabolite that is a common cause of some of the adverse events seen in patients treated with sulfasalazine (adverse events exemplified in the Background section of this application). In particular, patients with slow acetylation will benefit from reduced exposure to sulfapyridine. Furthermore, compliance with treatment may be improved due to the reduced burden of treatment through the use of fewer and / or smaller solid pharmaceutical compositions (e.g. tablets, microtablets, capsules, multi-unit pellet systems, etc.).

[0034] In the context of the present invention, the term "salt crystalline form of sulfasalazine of the present invention" refers to the organic acid crystal of 2-hydroxy-5-[2-[4-[(2-pyridinylamino)sulfonyl]phenyl]diazenyl]-benzoic acid (sulfasalazine) obtained by the preparation method of the present invention, and specifically exemplified is crystalline Form A of the D(-)-N-methylglucamine (meglumine) salt of sulfasalazine, crystalline Form A of the piperazine salt of sulfasalazine and crystalline Form B of the diethylamine salt of sulfasalazine.

[0035] In accordance with the present invention, the phrase "crystalline Form A of the D(-)-N-methylglucamine salt of sulfasalazine" may be used synonymously with "crystalline Form A of the meglumine salt of sulfasalazine," "meglumine salt Form A," "meglumine sulfasalazine Form A" or "sulfasalazine meglumine salt."

[0036] In accordance with the present invention, the phrase "crystalline Form A of the piperazine salt of sulfasalazine" may be used synonymously with "crystalline Form A of the piperazine salt of sulfasalazine," "piperazine salt Form A," "piperazine sulfasalazine Form A," or "sulfasalazine piperazine salt."

[0037] In accordance with the present disclosure, the phrase "crystalline Form A of the diethylamine salt of sulfasalazine" may be used synonymously with "crystalline Form A of the diethylamine salt of sulfasalazine," "diethylamine salt Form A," or "diethylamine sulfasalazine Form A." Crystalline Form A of the diethylamine salt of sulfasalazine, per se, is not covered by the present invention.

[0038] According to the present invention, the phrase "crystalline Form B of the diethylamine salt of sulfasalazine" may be used synonymously with "crystalline Form B of the diethylamine salt of sulfasalazine," "diethylamine salt Form B," or "diethylamine sulfasalazine Form B." According to any alternative embodiment of the present invention, crystalline Form B of the diethylamine salt of sulfasalazine is not covered by the present invention.

[0039] According to an optional alternative embodiment of the present invention, any of the diethylamine salts of sulfasalazine are not covered by the present invention.

[0040] According to one embodiment of all aspects of the invention, the sulfasalazine salt crystals of the invention are preferably at least 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% crystalline by weight, based on the total weight of each sulfasalazine salt form of the invention. Crystallinity may be estimated by conventional X-ray diffraction techniques.

[0041] According to a further embodiment of all aspects of the present invention, crystalline Form A of the meglumine salt of sulfasalazine exhibits at least the following characteristic powder X-ray diffraction (XPRD) peaks (expressed in 2θ°±0.2°) (in accordance with United States Pharmacopeia Convention. X-Ray Diffraction, General Test (USP941)): <941> United States Pharmacopeia, 25 th ed. Rockville, MD: United States Pharmacopeial Convention; 2002: 2088-2089): (1) 6.35, 13.93, and 22.41, or (2) 9.31, 15.86 and 20.99, or (3) 6.35, 13.93, 15.48, 15.86, 22.41, and 23.60; or (4) 6.35, 10.79, 12.93, 13.93, 15.48, 15.86, 18.12, 19.82, and 22.41; or (5) 9.31, 10.79, 12.93, 13.93, 14.47, 15.48, 15.86, 17.56, 19.10, 23.60, and 28.07; or (6) 6.35, 12.93, 13.93, 14.47, 15.48, 15.86, 19.10, 19.82, 20.99, 21.27, 22.41, 23.60, 23.89, and 28.07; or (7) 6.35, 9.31, 10.79, 12.93, 13.93, 14.47, 15.48, 15.86, 17.78, 18.12, 19.82, 20.11, 20.99, 21.27, 22.41, 23.60, 23.89, 28.07, and 28.80; or (8) 6.35, 9.31, 10.79, 12.93, 13.93, 14.47, 15.48, 15.86, 17.56, 17.78, 18.12, 18.50, 19.10, 19.82, 20.11, 20.99, 21.27, 22.41, 23.60, 23.89, 24.70, 25.14, 25.55, 25.93, 26.81, 28.07, and 28.80; or (9) 6.35, 9.31, 10.79, 12.93, 13.93, 14.47, 15.48, 15.86, 17.56, 17.78, 18.12, 18.50, 19.10, 19.82, 20.11, 20.99, 21.27, 22.41, 23.60, 23.89, 24.70, 25.14, 25.55, 25.93, 26.81, 28.07, 28.80, 29.49, 32.01, 32.58, 33.23.

[0042] According to a preferred embodiment of all aspects of the present invention, meglumine salt Form A exhibits at least the following characteristic XPRD peaks: 6.35, 13.93, 15.48, 15.86, 20.99, 22.41, 23.60 and 28.07.

[0043] FIG. 1 shows a characteristic XRPD spectrum of crystalline Form A of the meglumine salt of sulfasalazine of the present invention.

[0044] According to another or additional embodiment of any aspect of the invention, crystalline Form A of the piperazine salt of sulfasalazine exhibits at least the following characteristic X-ray powder diffraction (XPRD) peaks (expressed in 2θ°±0.2°) (in accordance with United States Pharmacopeia Convention. X-Ray Diffraction, General Test (USP941)): <941> United States Pharmacopeia, 25 th ed. Rockville, MD: United States Pharmacopeial Convention; 2002: 2088-2089): (1) 11.95, 12.30 and 16.42, or (2) December 30, December 93 and 15.01, or (3) 11.95, 12.30, 12.93, 16.42, 17.87, and 20.36; or (4) 8.11, 11.95, 12.30, 15.01, 16.42, 17.87, 20.36, and 20.74; or (5) 11.95, 12.30, 12.93, 15.01, 16.42, 17.87, 20.36, 20.74, 22.41, and 23.41; or (6) 11.95, 15.01, 16.42, 17.87, 20.36, 20.74, 23.41, 24.01, 24.67, 24.99, and 26.09; or (7) 8.11, 11.95, 12.30, 12.93, 15.01, 16.42, 17.87, 20.36, 20.74, 22.41, 23.41, 24.01, 24.67, 24.99, and 26.09, or (8) 11.95, 12.30, 12.93, 15.01, 16.42, 17.87, 20.36, 20.74, 22.41, 23.41, 24.01, 24.67, 24.99, 26.09, 26.81, 27.73, and 28.80; or (9) 8.11, 11.95, 12.30, 12.93, 15.01, 16.42, 17.87, 20.36, 20.74, 22.41, 23.41, 24.01, 24.67, 24.99, 26.09, 26.81, 27.73, 28.80, 29.80 and 30.43.

[0045] According to a preferred embodiment of all aspects of the present invention, piperazine salt Form A exhibits at least the following characteristic XPRD peaks: 12.30, 12.93, 15.01, 16.42, 22.41 and 23.41.

[0046] FIG. 2 shows a characteristic XRPD spectrum of crystalline Form A of the piperazine salt of sulfasalazine of the present invention.

[0047] The disclosure herein also provides that crystalline Form A of the diethylamine salt of sulfasalazine exhibits at least the following characteristic X-ray powder diffraction (XPRD) peaks (expressed in 2θ°±0.2°) (in accordance with United States Pharmacopeia Convention. X-Ray Diffraction, General Test (USP941)): <941> United States Pharmacopeia, 25 th ed. Rockville, MD: United States Pharmacopeial Convention; 2002: 2088-2089): (1) 7.16, 11.48, and 18.78, or (2) 10.50, 15.41, and 21.87; or (3) 7.16, 10.50, 11.48, 18.78, 21.65, and 21.87; or (4) 10.50, 11.48, 12.42, 14.38, 15.41, 16.64, 18.78, and 21.87; or (5) 7.16, 10.50, 11.01, 11.48, 13.87, 15.92, 16.64, 18.78, 21.08, 21.65, and 22.15; or (6) 7.16, 10.50, 11.01, 11.48, 12.42, 13.87, 14.38, 15.41, 15.92, 16.64, 17.19, 18.28, 18.78, 21.08, 21.65, 21.87, and 22.15; or (7) 7.16, 10.50, 11.48, 12.42, 13.87, 14.38, 15.41, 15.92, 16.64, 20.52, 21.08, 21.65, 21.87, 22.15, 22.47, 23.16, 23.63, 24.14, and 25.11, or (8) 7.16, 10.50, 11.01, 11.48, 12.42, 13.87, 14.38, 15.41, 15.92, 16.64, 17.19, 18.28, 18.78, 20.52, 21.08, 21.65, 21.87, 22.15, 22.47, 23.16, 23.63, 24.14, and 25.11, or (9) 7.16, 10.50, 11.01, 11.48, 12.42, 13.87, 14.38, 15.41, 15.92, 16.64, 17.19, 18.28, 18.78, 20.52, 21.08, 21.65, 21.87, 22.15, 22.47, 23.16, 23.63, 24.14, 25.11, 26.94, 27.95, 28.92, 29.46.

[0048] It is further disclosed that diethylamine salt Form A exhibits at least the following characteristic XPRD peaks: 7.16, 10.50, 11.48, 18.78, 21.65 and 21.87.

[0049] In FIG. 3, a characteristic XRPD spectrum of crystalline Form A of the diethylamine salt of sulfasalazine is shown.

[0050] According to still another or additional embodiment of any of the aspects of the present invention, the second polymorphic Form B crystalline of the diethylamine salt of sulfasalazine exhibits at least the following characteristic X-ray powder diffraction (XPRD) peaks (expressed in 2θ°±0.2°) (in accordance with United States Pharmacopeia Convention. X-Ray Diffraction, General Test (USP941)): <941> United States Pharmacopeia, 25 th ed. Rockville, MD: United States Pharmacopeial Convention; 2002: 2088-2089): (1) 6.85, 17.82, and 22.75, or (2) 11.38, 20.58, and 23.98, or (3) 6.85, 11.38, 17.62, 20.58, and 22.75; or (4) 6.85, 11.38, 11.70, 17.62, 20.58, 22.75, and 23.98; or (5) 11.38, 11.70, 15.29, 16.71, 17.62, 19.92, 20.58, 21.30, 22.75, 23.63, and 23.98; or (6) 6.85, 11.38, 11.70, 14.78, 15.29, 15.70, 16.71, 17.62, 19.92, 20.20, 20.58, 21.30, 22.75, 23.63, 23.98, and 28.61; or (7) 6.85, 11.38, 11.70, 14.78, 15.29, 15.70, 16.71, 17.62, 19.92, 20.20, 20.58, 21.30, 22.75, 23.63, 23.98, 25.05, 25.71, 26.81, 27.95, and 28.61; or (8) 6.85, 11.38, 11.70, 14.78, 15.29, 15.70, 16.71, 17.62, 19.92, 20.20, 20.58, 21.30, 22.75, 23.63, 23.98, 25.05, 25.71, 26.81, 27.51, 27.95, 28.61, 29.14, 31.06.

[0051] According to a preferred embodiment of all aspects of the present invention, diethylamine salt Form B exhibits at least the following characteristic XPRD peaks: 6.85, 11.38, 11.70, 17.62, 20.58, 22.75 and 23.98.

[0052] FIG. 4 shows a characteristic XRPD spectrum of crystalline Form B of the diethylamine salt of sulfasalazine of the present invention.

[0053] Although the sulfasalazine salt crystalline forms of the present invention, particularly Form A crystalline D(-)-N-methylglucamine (meglumine) salt of sulfasalazine, Form A crystalline piperazine salt of sulfasalazine and Form B crystalline diethylamine salt of sulfasalazine, can be used as solvates or hydrates, the present inventors have found that the sulfasalazine salt crystalline forms of the present invention can be obtained in solvate-free, particularly hydrate-free, forms by using the preparation method of the present invention. Such solvate-free, particularly hydrate-free (anhydrous) forms can exhibit advantageous physicochemical properties when preparing pharmaceutical compositions as solvate-free, particularly anhydrous forms, and in particular support the physical and chemical stability of the active ingredient sulfasalazine and the pharmaceutical composition, respectively, over the storage period.

[0054] Thus, the salt crystalline forms of sulfasalazine of the present invention, i.e., Form A crystalline of the D(-)-N-methylglucamine (meglumine) salt of sulfasalazine, Form A crystalline of the piperazine salt of sulfasalazine and Form B crystalline of the diethylamine salt of sulfasalazine, enable one skilled in the art to prepare stable pharmaceutical compositions, preferably stable pharmaceutical oral dosage formulations, more preferably stable pharmaceutical solid oral dosage formulations, each comprising sulfasalazine as an active ingredient. The term "stable" in the context of the present invention means that the measured values ​​fall within a particular range of values ​​as measured according to the respective applicable regulatory guidelines, e.g. the European Pharmacopoeia.

[0055] The properties or physical and chemical stability of the pharmaceutical tablet compositions of the invention may be tested in a conventional manner, for example by measuring the appearance, hardness (or crush resistance), disintegration time, dissolution, friability, moisture content, quantification of the sulfasalazine salt of the invention and / or its degradation products (related substances), and / or uniformity of dosage units or masses after storage under controlled storage conditions, such as intermediate and / or accelerated conditions according to ICH guideline Q1A(R2) (i.e., 25° C. / 60% relative humidity (RH) and / or 40° C. / 75% RH). These tests are performed in accordance with applicable pharmaceutical regulatory standards, for example as described in ICH or EMA guidelines and / or the European Pharmacopoeia (EP).

[0056] At least some of these attributes, i.e. properties or physical and chemical stability, of the pharmaceutical tablet composition of the present invention, preferably most of these attributes, most preferably all of these attributes are stable over time and over various controlled storage conditions. According to a preferred embodiment, the dissolution (profile) of the pharmaceutical tablet composition of the present invention, e.g. tablets or film-coated tablets according to the present invention, is stable for at least 6 months when stored at intermediate or long-term storage conditions, i.e. 25°C / 60% RH or 40°C / 75% RH, preferably in Alu-Alu blisters. More preferably, the dissolution and further attributes, e.g. quantification, related substances or dosage unit or mass uniformity, are also stable after storage for at least 6 months when stored at intermediate or long-term storage conditions.

[0057] Regarding the water vapour uptake capacity (rate and extent), the European Pharmacopoeia Technical guide 2015, General Chapter 5.11. classified solid pharmaceuticals into four different classes according to the degree of water uptake: slightly hygroscopic (uptake of ≥ 0.2% w / w but < 2% w / w at RH 80% and 25°C), hygroscopic (uptake of ≥ 2% w / w but < 15% w / w at RH 80% and 25°C), very hygroscopic (uptake of ≥ 15% w / w at RH 80% and 25°C) and deliquescent (absorbing enough water to form a liquid).

[0058] The inventors have further found that the sulfasalazine salt crystalline forms of the present invention, in particular the meglumine salt Form A, the piperazine salt Form A and / or the diethylamine salt Form B, are classified as only slightly hygroscopic materials. According to alternative or additional embodiments of all aspects of the present invention, the hygroscopicity of the sulfasalazine salt crystals of the present invention, preferably the meglumine salt Form A, is generally ≦1% w / w, more preferably ≦0.5% w / w water vapor uptake at 80% RH and 25° C. Thus, the sulfasalazine salt crystalline forms of the present invention are suitable for the preparation of stable pharmaceutical compositions.

[0059] According to a first aspect of the present invention, there is provided a process for preparing the organic salt crystalline form of sulfasalazine of the present invention.

[0060] According to step A1) of the process of the present invention, sulfasalazine free acid form is provided in a suitable solvent. According to another or additional embodiment of the present invention, the solvent of step A1) is generally selected from suitable organic solvents, preferably selected from the group consisting of acetone, acetonitrile and alcohols, such as methanol, ethanol, propanol, isopropanol and butanol.

[0061] According to step B1) of the process of the invention, an organic amine-containing component selected from meglumine and piperazine is provided in a suitable solvent. According to another or additional embodiment of the invention, the solvent in step B1) is preferably water or comprises water.

[0062] According to step C1) of the process of the present invention, the sulfasalazine solution of step A1) is mixed with the organic amine-containing component solution of step B1) at room temperature, preferably at 19° C. to 25° C. According to another or additional embodiment of the present invention, equimolar amounts of sulfasalazine free acid form and organic amine-containing component are used in step C1). According to yet another or additional embodiment of the present invention, the solutions of sulfasalazine and organic amine-containing component are mixed in step C1) at room temperature for at least 24 hours, preferably at least 48 hours, more preferably at least 72 hours, and even more preferably at least 96 hours.

[0063] According to step D1) of the preparation method of the present invention, the sulfasalazine salt crystalline form of the present invention, preferably sulfasalazine meglumine salt Form A crystal and sulfasalazine piperazine salt Form A crystal or solvates thereof formed in the solution in step C1) are separated. According to a preferred embodiment of all aspects of the present invention, the sulfasalazine salt crystalline form of the present invention is solvate-free, in particular anhydrous. In the context of the present invention, solvate-free and / or hydrate-free (anhydrous) means that the respective sulfasalazine salt crystalline form of the present invention contains ≦9 wt%, ≦5 wt%, ≦4 wt%, ≦3 wt%, ≦2 wt%, ≦1 wt% solvate and / or water based on the total weight of the respective sulfasalazine crystalline form of the present invention. If the sulfasalazine salt crystalline form of the present invention separated in step D1) contains a higher amount of residual solvate / water, an appropriate drying step may be carried out to make the sulfasalazine salt crystalline form of the present invention solvate-free and / or hydrate-free.

[0064] According to another method for preparing the salt crystalline form of sulfasalazine of the present invention, particularly sulfasalazine meglumine salt Form A crystalline form and sulfasalazine diethylamine salt Form B crystalline form of the present invention, the sulfasalazine free acid form is provided in a suitable solvent according to step A2), and an organic amine-containing component selected from meglumine and diethylamine is provided in a suitable solvent according to step B2). According to another or additional embodiment of the other method for preparation of the present invention, the solvent in steps A2) and B2) is generally independently selected from a suitable organic solvent, preferably selected from the group consisting of acetone, acetonitrile, and alcohols such as methanol, ethanol, propanol, isopropanol and butanol, more preferably the solvent in steps A2) and B2) is acetone.

[0065] According to another method for preparing the salt crystalline form of sulfasalazine of the present invention, the sulfasalazine solution in step A2) is mixed with the organic amine-containing component solution in step B2), where according to step C2) the amine-containing component is in at least a 5%, more preferably at least a 10%, more preferably at least a 20% molar excess relative to the sulfasalazine.

[0066] According to another method for preparing a salt crystalline form of sulfasalazine of the present invention, the solutions of sulfasalazine and the organic amine-containing component are preferably mixed in step C2) according to another or additional preparation method at an elevated temperature, preferably at a temperature in the range of 30° C. to 80° C., more preferably 40° C. to 65° C. According to yet another or additional embodiment of the another preparation method, the solutions of sulfasalazine and the organic amine-containing component are mixed in step C2) for up to 24 hours.

[0067] According to step D2a) of the alternative process for preparing the crystalline salt form of the present invention, the solution formed in step C2) is concentrated to isolate crystals of meglumine sulfasalazine Form A or a solvate thereof.

[0068] As an alternative to step D2a), according to step D2b) of another method for preparing the salt crystalline form of the present invention, an additional solvent may be added to the mixed solution formed in step C2), wherein the additional solvent is different from the solvent used in steps A2) and B2) to separate the crystals of meglumine sulfasalazine Form A or its solvate or the crystals of diethylamine sulfasalazine Form B or its solvate formed in step D2b). According to another or additional preferred embodiment, the solution containing the additional solvent in step D2b) is mixed at room temperature for at least 24 hours, preferably at a temperature in the range of 19°C to 25°C, preferably at room temperature for at least 48 hours, more preferably at room temperature for at least 60 hours. According to a further preferred embodiment, the solvent used in steps A2) and B2) is acetone and the additional solvent added during step D2b) is tert-butyl methyl ether.

[0069] If the crystalline salt form of sulfasalazine of the present invention isolated in step D2a) or D2b) contains a certain amount of residual solvate / water, an appropriate drying step may be performed to render the crystalline salt form of sulfasalazine of the present invention solvate-free and / or hydrate-free.

[0070] According to a third aspect of the present invention, there is provided a pharmaceutical composition comprising a therapeutically effective amount of one or more of the crystalline salt forms of sulfasalazine of the present invention, particularly as described above or in the Examples. The pharmaceutical composition of the present invention may comprise one, two, three or more pharma- ceutical acceptable adjuvants in addition to one or more of the crystalline salt forms of sulfasalazine of the present invention.

[0071] Depending on the mode of administration, for example, the pharmaceutical composition of the present invention may contain 0.01 to 100% by weight, 1 to 90% by weight, 25 to 80% by weight, 30 to 70% by weight, 40 to 60% by weight, or 50% by weight of the crystalline salt form of sulfasalazine of the present invention, based on the total weight of the pharmaceutical composition of the present invention.

[0072] The pharmaceutical composition of the present invention may be administered systemically, for example, orally in the form of tablets, microtablets, granules, powders, capsules, syrups or multi-unit pellet systems (also called MUPS); or parenterally (e.g., intravenously, subcutaneously, intraarticularly) in the form of solutions or suspensions; or rectally in the form of suppositories, foams, etc. Preferably, the pharmaceutical composition of the present invention is administered orally.

[0073] When the pharmaceutical composition of the present invention is administered orally, the sulfasalazine salt crystalline form of the present invention in the pharmaceutical composition of the present invention is preferably protected from contact with acidic gastric fluids, for example by an enteric coating layer provided on or within the pharmaceutical composition of the present invention.

[0074] According to another embodiment of the pharmaceutical composition of the present invention, one or more of the sulfasalazine salt crystalline forms of the present invention are mixed with one, two, three, four or more pharmaceutical tableting additives, preferably selected from the group consisting of excipients, binders, disintegrants, lubricants, etc., and compressed into a tableted dosage form.

[0075] The compressed tablet of the present invention is optionally covered / coated with one or more film-forming agents, which may include, for example, alkaline substances, to obtain a smooth surface of the tablet and further improve the stability of the tablet during packaging, transportation and storage. Alternatively or additionally, such tablet coating layer may include additives such as coloring agents and pigments to obtain a good appearance of the tablet. Alternatively or additionally, the tablet composition of the present invention may include an enteric coating layer that protects the salt crystal form of sulfasalazine of the present invention from contact with gastric acid. Alternatively or additionally, the tablet composition of the present invention may include pharmaceutical additives that facilitate the immediate or sustained release of the salt crystal form of sulfasalazine of the present invention. In particular, the tablet coating may be a cellulose derivative, such as pregelatinized starch, a cellulose ether (e.g. ethyl cellulose (EC), methyl cellulose (MC), hydroxyethyl cellulose (HEC) or hydroxypropyl cellulose (HPC); in particular crosslinked sodium carboxymethylcellulose) or an ester or semiester of cellulose (e.g. cellulose acetate phthalate (CAP) or hydroxypropylmethylcellulose phthalate (HPMCP)); an acrylic acid polymer or copolymer, preferably a methacrylate aminoester copolymer (e.g. Eudragit RS or Eudragit RL) or a methacrylate copolymer (e.g. methacrylate copolymer). The composition may comprise one, two, three, four, five or more components selected from the group consisting of methyl acrylate-ethyl acrylate copolymers (e.g. methyl methacrylate-ethyl acrylate copolymer 1:1); waxy substances (e.g. carnauba wax); polyethylene glycols (e.g. macrogol 6,000, macrogol 20,000); (crosslinked) polyvinylpyrrolidones (e.g. povidone K30, povidone K25, crospovidone), polyvinyl alcohols or derivatives such as polyvinyl acetate phthalate (PVAP); pigments (e.g. titanium dioxide); stearic acid, magnesium stearate or glycerol monostearate; and talc.

[0076] According to another embodiment of the pharmaceutical composition of the invention for oral administration, the composition of the invention is alternatively or additionally suitable for dispersion in an aqueous liquid having a neutral or slightly acidic pH prior to oral administration or feeding via a nasogastric tube.

[0077] According to yet another embodiment of the pharmaceutical composition of the present invention for oral administration, the pharmaceutical composition of the present invention may be provided in the form of a hard or soft capsule, preferably a soft gelatin capsule, in which the sulfasalazine salt crystalline form of the present invention may be mixed, for example, with vegetable oil or polyethylene glycol. Also, the liquid or semi-solid formulation of the sulfasalazine salt crystalline form of the present invention may be filled into a hard gelatin capsule to form the composition of the present invention.

[0078] A single unit dose of the pharmaceutical composition of the present invention in all aspects of the present invention for oral administration may generally contain one or more of the crystalline salt forms of sulfasalazine of the present invention in the range of 1 mg to 2000 mg.In particular, the pharmaceutical composition of the present invention for oral administration may contain, per unit, 10 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 75 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 125 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 175 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 225 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 275 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 325 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 375 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 510 mg, 520 mg, 530 mg, 540 mg, 550 mg, 560 mg, 570 mg, 580 mg, 590 mg, 600 mg, 610 mg, 620 mg, 630 mg, 640 mg, 650 mg, 660 mg, 670 mg, 680 mg, 690 mg, 700 mg, 710 mg, 720 mg, 730 mg, 740 mg, 750 mg, 760 mg mg, 420 mg, 425 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 475 mg, 480 mg, 490 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, 800 mg, 825 mg, 850 mg, 875 mg, 900 mg, 925 mg, 950 mg, 975 mg, 1,000 mg, 1,025 mg, 1,050 mg, 1,075 mg, 1,100 mg, 1,125 mg, 1,150 mg, 1,175 mg, 1,200 mg, 1,225 mg, 1,250 mg, 1,275 mg, 1,300 The sulfasalazine salt crystalline forms of the present invention may comprise one or more of the sulfasalazine salt crystalline forms of the present invention in amounts of 1,325 mg, 1,350 mg, 1,375 mg, 1,400 mg, 1,425 mg, 1,450 mg, 1,475 mg, 1,500 mg, 1,525 mg, 1,550 mg, 1,575 mg, 1,600 mg, 1,625 mg, 1,650 mg, 1,675 mg, 1,700 mg, 1,725 ​​mg, 1,750 mg, 1,775 mg, 1,800 mg, 1,825 mg, 1,850 mg, 1,875 mg, 1,900 mg, 1,925 mg, 1,950 mg, 1,975 mg, or 2,000 mg.Preferably, the amount of one or all of the sulfasalazine salt crystalline forms of the present invention is less than a single unit dose of a comparative pharmaceutical composition containing sulfasalazine free acid alone. Thus, for oral dosage forms (e.g., tablets, microtablets, granules, powders, capsules, multi-unit pellet systems) of the pharmaceutical composition of the present invention, a single unit dose preferably contains <500 mg, ≦450 mg, ≦400 mg, ≦375 mg, ≦250 mg of the sulfasalazine salt crystalline forms of the present invention.

[0079] Alternatively, the pharmaceutical composition of the present invention may be in the form of a liquid preparation for oral administration in the form of a syrup or suspension.The pharmaceutical additives contained in such liquid preparations may include sugar and / or a mixture of ethanol, water, glycerol and propylene glycol, preferably buffered to an appropriate pH.If desired, such liquid preparations of the present invention may include one, two, three, four or more additional additives, preferably selected from the group consisting of coloring agents, flavoring agents, saccharin and / or carboxymethylcellulose as a thickening agent or other additives known to those skilled in the art.

[0080] According to yet another or additional embodiment of the pharmaceutical composition of the present invention, the pharmaceutical composition preferably comprises a non-steroidal anti-inflammatory drug; preferably a non-selective cyclo-oxygenase COX-1 / COX-2 inhibitor, whether applied topically or systemically, such as piroxicam, diclofenac, propionic acids, such as naproxen, flurbiprofen, fenoprofen, ketoprofen and ibuprofen, fenamates, such as mefenamic acid, indomethacin, sulindac, azapropayone, pyrazolones, such as phenylbutazone, salicylates, such as aspirin, selective COX-2 inhibitors, such as meloxicam, celecoxib, rofecoxib, valdecoxib, lumiracoxib, sulindac ... coxib), parecoxib and etoricoxib, cyclooxygenase inhibitors with nitric oxide donating activity (CINOD); glucocorticoids, preferably flunisolide, triamcinolone acetonide, betamethasone dipropionate, budesonide, fluticasone propionate, ciclesonide or mometasone furoate; methotrexate; leflunomide; hydroxychloroquine; d-penicillamine; diacerein; dietary supplements, preferably glucosamine; gold preparations, preferably auranofin; cytokines or agonists or antagonists of cytokine function; monoclonal antibodies targeting B lymphocytes, preferably CD20 (rituximab); MRA-aIL16R; T-lymphocytes; CTLA4-Ig; HuMax 11-15; modulators of chemokine receptor function, preferably antagonists of CCR2, CCR2A, CCR2B, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCRlO and CCR11 (CC family), CXCRl, CXCR2, CXCR3, CXCR4 and CXCR5 (CXC family) and CX3CRl (C-X3-C family); azathioprine, tofacitinib, monoclonal antibodies such as the anti-tumor necrosis factor alpha monoclonal antibodies infliximab, adalimumab and golimumab; interleukin 1 receptor antagonists such as anakinra; etanercept and abatacept;More preferably, it may further comprise one, two, three or more additional active ingredients selected from the group consisting of methotrexate and hydroxychloroquine. Alternatively or in addition, the pharmaceutical composition of the present invention may comprise sulfasalazine free acid form. Preferably, the amount of sulfasalazine free acid form is less than the amount of the salt crystalline form of sulfasalazine of the present invention in the pharmaceutical composition of the present invention.

[0081] The salt crystalline forms of sulfasalazine of the present invention and its in vivo metabolites sulfapyridine and 5-ASA are useful as modulators of the function of various inflammatory cell types, such as T cells, B cells, dendritic cells, neutrophils, NK cells and mast cells.For example, in experiments studying the proliferation of human synovial cells from patients with rheumatoid arthritis, the proliferation of these cells and the production of IL-1B and IL-6 by these cells were shown to be significantly inhibited.In these experiments, it could be shown that the overexpression of c-fos mRNA is inhibited by the crystalline forms of sulfasalazine of the present invention.Therefore, the crystalline forms of sulfasalazine of the present invention can be administered to mammals, including humans, for the treatment of, in particular, autoimmune, inflammatory, proliferative and hyperproliferative diseases and immune-mediated diseases.

[0082] Thus, according to another embodiment of all aspects of the invention, the pharmaceutical compositions of the invention and the crystalline forms of the invention comprise: i) diseases or conditions in which modulation of inflammatory cells is beneficial; ii) diseases or conditions relating to bone or joints, preferably arthritis associated with or including osteoarthritis, primary or secondary to, for example, congenital hip dysplasia; cervical and lumbar spondylitis, and lower back and neck pain; rheumatoid arthritis and Still's disease; seronegative spondyloarthropathy, such as ankylosing spondylitis, psoriatic arthritis, reactive arthritis and undifferentiated spondarthropathy, septic arthritis and other arthopathies and bone disorders associated with infection, such as tuberculosis, e.g. Pott's disease and Ponce's disease; acute and chronic crystal-induced synovitis, such as uric acid gout, calcium pyrophosphate deposition disease, and calcium apatite-related tendon, bursa and synovial inflammation; Behcet's disease; primary or secondary Sjogren's syndrome. systemic sclerosis and localized scleroderma; systemic lupus erythematosus, mixed connective tissue disease, and undifferentiated connective tissue disease; inflammatory myopathies, e.g., dermatomyositis and polymyositis; polymyalgia rheumatica; juvenile arthritis, e.g., idiopathic inflammatory arthritis of any joint location and associated syndrome, and rheumatic fever and its systemic complications; vasculitis, e.g., giant cell arteritis, Takayasu's arteritis, Churg-Strauss syndrome, polyarteritis nodosa, microscopic polyarteritis, and vasculitis associated with viral infections, hypersensitivity reactions, cryoglobulins, and paraproteins; lower back pain; Familial Mediterranean Fever, Muckle-Wells syndrome, and Familial Hibernian Fever, Kikuchi's disease; drug-induced arthralgia, tendonitis, and myopathy; and iii) diseases or conditions relating to the digestive tract, preferably selected from the group consisting of eosinophilic gastroenteritis, mastocytosis, Crohn's disease, colitis, e.g. ulcerative colitis, proctitis; celiac disease, irritable bowel syndrome, and food-related allergies that may affect areas distant from the intestine, e.g. migraine, rhinitis or eczema. The invention is for use in the treatment of

[0083] According to another aspect, the present invention relates to the use of a crystalline salt form of sulfasalazine according to the invention or a pharmaceutical composition according to the invention in the manufacture of a medicament for / treatment of the above mentioned disorders or conditions.

[0084] According to another aspect, the present invention relates to a method for treating one or more of the above disorders or conditions in a patient suffering from or at risk of said disease or condition, comprising administering to the patient a therapeutically effective amount of a crystalline salt form of sulfasalazine of the present invention or a pharmaceutical composition of the present invention.

[0085] According to all aspects, the present invention also relates to combination therapy in which one or more crystalline salt forms of sulfasalazine of the present invention or pharmaceutical compositions of the present invention are administered concurrently (simultaneously) or sequentially or as a combined pharmaceutical formulation or combined dosing regimen with one or more active ingredients (therapeutic agents) for the treatment of one or more of the diseases and conditions, preferably the above mentioned diseases and conditions.

[0086] In the context of this specification, the term "treatment" also includes "prophylaxis", unless specifically stated to the contrary. The terms "therapeutic" and "therapeutically" should be construed accordingly.

[0087] Prevention is expected to be particularly relevant to the treatment of patients suffering from, or otherwise considered to be at high risk of, the disease or condition in question. Those at risk of developing a particular disease or condition generally include those who have a family history of the disease or condition, or those who have been identified by genetic testing or screening to be predisposed to developing the disease or condition.

[0088] According to the inventive treatment of the above inflammatory diseases, one, two, three, four or more of the inventive salt crystalline forms of sulfasalazine or the inventive pharmaceutical compositions may be used in the same or separate pharmaceutical composition with one, two, three or more active ingredients (therapeutic agents), which are preferably selected from the group consisting of the following therapeutic agents: - nonsteroidal anti-inflammatory drugs (NSAIDs), for example nonselective cyclo-oxygenase COX-1 / COX-2 inhibitors, whether applied topically or systemically (e.g. piroxicam, diclofenac, propionic acids, e.g. naproxen, flurbiprofen, fenoprofen, ketoprofen and ibuprofen, fenamates, e.g. mefenamic acid, indomethacin, sulindac, azapropazones, pyrazolones, e.g. phenylbutazone, salicylates, e.g. aspirin); selective COX-2 inhibitors (e.g. meloxicam, diclofenac, propionic acids, e.g. naproxen, flurbiprofen, fenoprofen, ketoprofen and ibuprofen); cicam, celecoxib, rofecoxib, valdecoxib, lumiracoxib, parecoxib and etoricoxib); cyclooxygenase inhibitors that donate nitric oxide (CINOD); glucocorticoids (whether administered by topical, oral, intramuscular, intravenous or intra-articular routes); methotrexate, leflunomide, hydroxychloroquine, d-penicillamine; auranofin and other parenteral or oral gold preparations; analgesics; diacerein; intra-articular treatments, such as hyaluronic acid derivatives; and dietary supplements, such as glucosamine. - agonists or antagonists of cytokines or cytokine function (including drugs acting on cytokine signaling pathways, e.g. modulators of the SOCS system), such as α-, β- and γ-interferon; insulin-like growth factor type I (IGF-I); interleukins (IL), such as IL-1 to 23, and interleukin antagonists or inhibitors, such as anakinra; tumor necrosis factor alpha (TNF-α) inhibitors, such as anti-TNF monoclonal antibodies (e.g. infliximab, adalimumab and golimumab) and TNF receptor antagonists, such as immunoglobulin molecules (e.g. etanercept) and low molecular weight drugs, such as pentoxifylline. - Monoclonal antibodies targeting B-lymphocytes (e.g. CD20 (rituximab); MRA-aIL16R and T-lymphocytes, CTLA4-Ig; HuMax 11-15). - modulators of chemokine receptor function, such as CCR2, CCR2A, CCR2B, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCRlO and CCR11 (CC family); antagonists of CXCRl, CXCR2, CXCR3, CXCR4 and CXCR5 (CXC family) and the CX3CRl C-X3-C family. - matrix metalloproteases (Tv-IMP), i.e. stromelysin, collagenase and gelatinase, as well as aggrecanase; in particular collagenase-1 (MMP-1), collagenase-2 (MMP-8), collagenase-3 (MMP-13), stromelysin-1 (MMP-3), stromelysin-2 (MMP-10), and stromelysin-3 (MMP-11) and inhibitors of MMP-9 and MMP-12, such as doxycycline. - leukotriene biosynthesis inhibitors, 5-lipoxygenase (5-LO) inhibitors or 5-lipoxygenase-activating protein (FLAP) antagonists, such as; zileuton; ABT-761; fenleuton; tepoxalin; Abbott-79175; Abbott-85761; N-(5-substituted)-thiophene-2-alkylsulfonamides; 2,6-di-tert-butylphenol hydrazones; methoxytetrahydropyrans, such as Zeneca ZD-213S; compound SB-210661; pyridinyl-substituted 2-cyanonaphthalene compounds, such as L-739,010; 2-cyanoquinoline compounds, such as L-746,530; or indole or quinoline compounds, such as MK-591, MK-886, and BAYx1005. - Leukotriene (LT) B4, LTC4, LTD4 and LTE4 receptor antagonists selected from the group consisting of phenothiazine-3-ls, such as L-651,392; amidino compounds, such as CGS-25019c; benzoxalamines, such as ontazolast; and compounds such as zafiruukast, ablukast, montelukast, pranlukast, verlukast (MK-679), RG-12525, Ro-245913, iralukast (CGP 45715A), and BAYx7195. - phosphodiesterase (PDE) inhibitors, such as methylxanthanines, e.g. theophylline and aminophylline; selective PDE isoenzyme inhibitors, such as PDE4 inhibitors, inhibitors of the isoform PDE4D, e.g. apremilast or inhibitors of PDE5. - Endothelin antagonists, such as Tezosentan, Bosentan, Macitentan, Enrasentan and Sixtasentan. - Angiotensin II antagonists, such as azilsartan, losartan, valsartan, candesartan and telmisartan. - Dual antagonists against both the angiotensin II and endothelin A receptors (DARAs), as disclosed, for example, in WO2000001389 and WO2001044239. - an adenosine A2a agonist, such as CGS-21680, and / or an adenosine A3 agonist, such as IB-MECA, and / or an adenosine A2b antagonist.The present invention further relates to a combination of a compound of the invention with a histamine type 1 receptor antagonist, such as cetirizine, loratadine, desloratadine, fexofenadine, acrivastine, terfenadine, astemizole, azelastine, levocabastine, chlorpheniramine, promethazine, cyclizine or mizolastine, applied orally, topically or parenterally. - Proton pump inhibitors (e.g. omeprazole) or gastroprotective histamine type 2 receptor antagonists. - Histamine type 4 receptor antagonist. - alpha1 / alpha2 adrenergic receptor agonist vasoconstrictor sympathomimetics, such as propylhexedrine, phenylephrine, phenylpropanolamine, ephedrine, pseudoephedrine, naphazoline hydrochloride, oxymetazoline hydrochloride, tetrahydrozoline hydrochloride, xylometazoline hydrochloride, tramazoline hydrochloride or ethylnorepinephrine hydrochloride. - anticholinergics, for example muscarinic receptor (M1, M2 and M3) antagonists, such as atropine, hyoscine, glycopyrrolate, ipratropium bromide, tiotropium bromide, oxitropium bromide, pirenzepine or telenzepine. - beta-adrenergic receptor agonists (including beta-receptor subtypes 1-4), such as isoprenaline, salbutamol, formoterol, salmeterol, terbutaline, orciprenaline, bitolterol mesylate or pirbuterol, or their chiral enantiomers. - chromones, such as sodium cromoglycate or nedocromil sodium.The invention still further relates to combinations of the compounds of the invention with glucocorticoids, such as flunisolide, triamcinolone acetonide, betamethasone dipropionate, budesonide, fluticasone propionate, ciclesonide or mometasone furoate. - Drugs that modulate nuclear hormone receptors, e.g. PPARs. - Immunoglobulins (Ig) or Ig preparations or antagonists or antibodies that modulate Ig function, such as anti-IgE (e.g. omalizumab). - Another systemically or topically applied anti-inflammatory drug, such as thalidomide or its derivatives, retinoids, dithranol or calcipotriol. - aminosalicylates and sulfapyridines, such as sulfasalazine, mesalazine, balsalazide and olsalazine; and immunomodulators, such as thiopurines, and corticosteroids, such as budesonide. Preferably, the amount of further aminosalicylates and sulfapyridines is less than the amount of the sulfasalazine crystalline form of the invention per unit dosage form. - antibacterial drugs, such as penicillin derivatives, tetracyclines, macrolides, beta-lactams, fluoroquinolones, metronidazole, inhaled aminoglycosides; antiviral drugs, such as acyclovir, famciclovir, valacyclovir, ganciclovir, cidofovir, amantadine, rimantadine, ribavirin, zanamavir and oseltamavir; protease inhibitors, such as indinavir, nelfinavir, ritonavir and saquinavir; nucleoside reverse transcriptase inhibitors, such as didanosine, lamivudine, stavudine, zalcitabine or zidovudine; or non-nucleoside reverse transcriptase inhibitors, such as nevirapine or efavirenz. - cardiovascular drugs, such as calcium channel blockers, beta-adrenergic receptor blockers, angiotensin-converting enzyme (ACE) inhibitors, angiotensin 2 receptor antagonists; lipid-lowering drugs, such as statins or fibrates; modulators of blood cell morphology, such as pentoxifylline; thrombolytic or anticoagulant drugs, such as platelet aggregation inhibitors. - antidepressants (e.g. sertraline), anti-Parkinson's drugs (e.g. deprenyl, L-dopa, ropinirole, pramipexole, MAOB inhibitors such as selegiline and rasagiline, comP inhibitors such as tasmar, A-2 inhibitors, dopamine reuptake inhibitors, NMDA antagonists, nicotinic agonists, dopamine agonists or inhibitors of neuronal nitric oxide synthase), or anti-Alzheimer's drugs such as donepezil, rivastigmine, tacrine, COX-2 inhibitors, propentofylline or metrifonate. - Medications for the treatment of acute or chronic pain, such as centrally or peripherally acting analgesics (e.g. opioids or derivatives thereof), carbamazepine, phenytoin, sodium valproate, amitryptiline or other antidepressants, paracetamol, or nonsteroidal anti-inflammatory drugs. - Parenterally or topically applied (including inhaled) local anaesthetics, such as lignocaine or its derivatives. - Anti-osteoporosis drugs, for example hormonal agents, such as raloxifene, or biphosphonates, such as alendonic acid. - one or more drugs selected from the group consisting of: (i) tryptase inhibitors; (ii) platelet activating factor (PAF) antagonists; (iii) interleukin converting enzyme (ICE) inhibitors; (iv) IMPDH inhibitors; (v) adhesion molecule inhibitors, e.g. VLA-4 antagonists; (vi) cathepsins; (vii) kinase inhibitors, e.g. inhibitors of tyrosine kinases (e.g. Btk, Itk, Jak3 or MAP, e.g. tofacitinib, gefitinib or imatinib mesylate), serine / threonine kinases ( For example, inhibitors of MAP kinases, e.g. p38, INK, protein kinase A, B or C, or inhibitors of κB kinases, e.g. IKK1, IKK2 or IKK3, or kinases involved in cell cycle control (e.g. cyclin-dependent kinases); (viii) glucose-6-phosphate dehydrogenase inhibitors; (ix) kyrin-B1 or -B2-receptor antagonists; (x) anti-gout drugs, e.g. colchicine; (xi) xanthine oxidase inhibitors, e.g. allopurinol; (xii) uricosurics. (xiii) growth hormone secretagogues; (xiv) transforming growth factor (TGFβ); (xv) platelet-derived growth factor (PDGF); (xvi) fibroblast growth factors, such as basic fibroblast growth factor (bFGF); (xvii) granulocyte-macrophage colony-stimulating factor (GM-CSF); (xviii) capsaicin cream; (xix) tachykinin NK1 or NK3 receptor antagonists, such as NKP-608C, SB -233412 (talnetant) or D-441S; (xx) elastase inhibitors, such as LT-77 or ZD-0892; (xxi) TNF-α converting enzyme inhibitors (TACE); (xxii) inducible nitric oxide synthase (iNOS) inhibitors; (xxiii) chemoattractant receptor homologues expressed in TH2 cells (e.g. CRTH2 antagonists); (xxiv) inhibitors of P38; (xxv) drugs that modulate the function of Toll-like receptors (TLRs), (xxvi) drugs that modulate the activity of purinergic receptors such as P2X7;or (xxvii) inhibitors of transcription factor activation such as NFkB, API or STATS, (xxvii) drugs that modulate guanylate cyclase, such as riociguat (methyl-N-[4,6-diamino-2-[1-[(2-fluorophenyl)methyl]-1H-pyrazolo[3,4-b]pyridin-3-yl]-5-pyrimidinyl]-N-methyl-carbaminate (IUPAC)); - one or more therapeutic agents for the treatment of cancer, preferably (i) antiproliferative / antineoplastic agents or combinations thereof used in clinical oncology, such as alkylating agents (e.g. cisplatin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulfan or nitrosoureas); antimetabolites (e.g. antifolates, such as fluoropyrimidines like 5-fluorouracil, or tegafur, raltitrexed, methotrexate, cytosine arabinoside, hydrochloride, cyclosporine, cyclopentasiloxane, cyclohexyl tert-butyl ether ... xyluria, gemcitabine or paclitaxel; antitumor antibiotics (e.g. anthracyclines, such as adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin or mithramycin); mitotic inhibitors (e.g. vinca alkaloids, such as vincristine, vinblastine, vindesine or vinorelbine, or taxoids, such as taxol or taxotere); or topoisomerase inhibitors (e.g. epipodophyllotoxins, e.g. etoposide, teniposide, amsacrine, topotecan or camptothecin); (ii) cytostatics, e.g. antiestrogens (e.g. tamoxifen, toremifene, raloxifene, droloxifene or iodoxyfene), estrogen receptor downregulators (e.g. fulvestrant), antiandrogens (e.g. bicalutamide, phytamide, nilutamide or cyproterone acetate), LHRH antagonists or LHRH antagonists. (iii) drugs that inhibit the invasion of cancer cells (e.g. metalloproteinase inhibitors like marimastat, or inhibitors of urokinase plasminogen activator receptor function);(iv) inhibitors of growth factor function, for example: growth factor antibodies (e.g. the anti-erbb2 antibody trastuzumab or the anti-erbb1 antibody cetuximab [C225]), farnesyltransferase inhibitors, tyrosine kinase inhibitors or serine / threonine kinase inhibitors, inhibitors of the epidermal growth factor family (e.g. EGFR family tyrosine kinase inhibitors, for example N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine (gefitinib, AZDl 839), N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine (erlotinib, OSI-774) or 6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)quinazolin-4-amine (CI 1033), inhibitors of the platelet derived growth factor family, or inhibitors of the hepatocyte growth factor family; (v) antiangiogenic agents, such as those that inhibit the action of vascular endothelial growth factor (e.g. the anti-vascular endothelial growth factor antibody bevacizumab, compounds disclosed in WO 97 / 22596, WO 97 / 30035, WO 97 / 32856 or WO 98 / 13354), or compounds acting by another mechanism (e.g. linomide, inhibitors of integrin αvβ3 function or angiostatin); (vi) vascular disorder agents, such as combretastatin A4, or compounds disclosed in WO 99 / 02166, WO 00 / 40529, WO 00 / 41669, WO 01 / 92224, WO 02 / 04434 or WO 02 / 08213; (vii) drugs used in antisense therapy, such as those directed against one of the above targets, e.g. ISIS 2503, anti-ras antisense; (viii) gene therapy approaches, such as approaches to replace abnormal genes, e.g. abnormal p53 or abnormal BRCA1 or BRCA2, GDEPT (gene directed enzyme prodrug therapy) approaches, such as those using cytosine deaminase, thymidine kinase or bacterial nitroreductase enzymes, and approaches to increase a patient's resistance to chemotherapy or radiotherapy, such as drugs used in multidrug resistance gene therapy;or (ix) a therapeutic agent selected from the group consisting of drugs used in immunotherapy approaches, such as ex vivo and in vivo approaches to increase the immunogenicity of patient tumor cells, such as transfection of cytokines, such as interleukin 2, interleukin 4 or granulocyte-macrophage colony stimulating factor, approaches to reduce T cell anergy, approaches using transfected immune cells, such as cytokine-transfected dendritic cells, approaches using cytokine-transfected tumor cell lines and approaches using anti-idiotypic antibodies; - one or more therapeutic agents for the treatment of airway disease, respiratory disease and / or inflammatory disease, such as chronic obstructive pulmonary disease and asthma. The sulfasalazine salt crystal form of the present invention can be administered by inhalation or oral route, and the drug (other drug) for the treatment of airway disease, respiratory disease and / or inflammatory disease, such as chronic obstructive pulmonary disease and asthma, can be independently selected to be administered by inhalation or oral route. The sulfasalazine salt crystal form of the present invention and other drugs can be administered in one pharmaceutical preparation or in separate pharmaceutical preparations. In the case of separate pharmaceutical preparations, the sulfasalazine salt crystal form of the present invention and other drugs can be administered simultaneously or sequentially or separately.

[0089] Depending on the therapeutic application, the dosage of the sulfasalazine salt crystalline forms of the present invention will of course vary depending on the mode of administration, the desired treatment and the intended disorder, but may typically range from 1 mg / kg to 50 mg / kg. EXAMPLES

[0090] The present invention is described below on the basis of exemplary embodiments, which serve merely as examples and do not limit the scope of the present protection. The exemplified features may be combined separately or in any (sub)combination with the general disclosure of all aspects of the invention above.

[0091] General method

[0092] 1 H-NMR and13 C-NMR The produced examples were subjected to proton and carbon nuclear magnetic resonance ( 1 H-NMR and 13 The compounds were characterized using C-NMR for identity and salt stoichiometry, X-ray powder diffraction (XRPD) for crystallinity changes, differential scanning calorimetry (DSC) for thermal properties, vapour sorption gravimetry (GVS) for water interactions, salt stoichiometry (HPLC), counterion identity (CE) and finally solubility in water (HPLC).

[0093] 1 H NMR and 13 C NMR spectra were recorded on a Varian Unity Inova 400 MHz (software: VKMR 6.1C and VNMRJ 1.1D; probe: Xalorac 5mm DG400-5AT) or Varian Mercury - VX 300 MHz (software: VNMR 6.1C; probe: Varian 5mm AutoSW PFG) instrument at 298 K. The central peak of acetone-d6 or dimethylsulfoxide (DMSO)-d6 was used as an internal standard.

[0094] XRPD X-ray powder diffraction (XRPD) analysis may be performed on samples prepared according to standard methods (see, e.g., Giacovazzo et al., eds., Fundamentals of Crystallography, Oxford University Press (1992); Jenkins & Snyder, eds., Introduction to X-Ray Powder Diffractometry, John Wiley & Sons, New York (1996); Bunn, ed., Chemical Crystallography, Clarendon Press, London (1948); and Klug and Alexander, eds., X-Ray Diffraction Procedures, John Wiley & Sons, New York (1974)). The precipitated samples were smeared onto a zero background sample holder and measured from 2-35° (2-theta) using a Thermo ARL X'tra diffractometer equipped with a Peltier-cooled solid-state detector, Cu tube (λ=1.5418 Å), 45 kV / 44 mA, using a rotating sample holder and continuous scanning with a scan rate of 2° / min and a step size of 0.02°. The standard deviation is ±0.2° 2-theta (2θ).

[0095] DSC Differential scanning calorimetry (DSC) was performed using standard methods, e.g., as described in Hoehne, GWH et al (1996), Differential Scanning Calorimetry, Springer, Berlin, and the calometric response of the test samples to increasing temperature was examined using a PerkinElmer Pyris DSC. Temperature sensations were typically between 60 and 285°C, but were slightly varied depending on the results and the need for remeasurements. The scan rate was 10°C / min. Approximately 2 mg of sample was used. Measurements were performed using open aluminum pans and a dry nitrogen atmosphere to avoid oxidative degradation. It is well known that the onset and peak temperatures of DSC can vary depending on the purity of the sample and the instrument parameters, especially the temperature scan rate. Those skilled in the art can use routine optimization / calibration to set the DSC instrument parameters so that data comparable to those presented herein can be collected.

[0096] GVS Gravimetric vapour sorption (GVS) was used to determine the hygroscopicity of the samples: Experiments were carried out at 25°C using a DVS 1 instrument from SMS Ltd, and adsorption / desorption isotherms were recorded using various methods, the main features being: one adsorption / desorption cycle was performed from 0% RH to 80% RH, with 10% RH steps, a dm / dt trigger value of 0.002% (dm / dt = change in mass with time, when the balance stability is in this range, the next step is automatically started, but if these conditions are not achieved, the default maximum time is 6 hours for each step). The amount of sample used was 1-3 mg.

[0097] HPLC The amount of dissolved sulfasalazine was determined by HPLC on an Agilent 1100 instrument using a Waters XTerra 3.5 μm C18 column (50×4.6 mm) and a mobile phase consisting of 95% ethanol / 25 mM phosphoric acid, 45 / 55. The flow rate was 1.0 mL / min, the injection volume was 5 μL, and the detection wavelengths were 358 nm (for assay) and 260 nm (for chromatographic purity). Quantitation was performed using an external standard method. The quantitative method was validated for selectivity, reproducibility and linearity.

[0098] CE In capillary electrophoresis (CE), the positive identity of selected counter ions was indicated by the corresponding migration times between the peaks of the sample and standard solutions. The instrument used was a Hewlett-Packard 3D-CE. The capillary was fused silica with an inner diameter of 50 μm and an effective length of 56 cm. The electrolyte was Agilent Cation Buffer for CE (PN 5064-8203), the potential was +30 kV, and the injection was 50 mbar x 10 s (sample solution) and 25 mbar x 3 s (standard solution). Detection was performed using a UV detector at 310 nm with a reference wavelength of 215 nm.

[0099] TGA Thermogravimetric analysis (TGA) equipment: PerkinElmer TGA7 Method: Approximately 3 mg of sample was placed in an open Pt-pan, weighed, and analyzed using a scan rate of 10°C / min from 22°C to 120°C, followed by a 30 minute hold at 120°C in a dry nitrogen gas flow to ensure an inert atmosphere.

[0100] Example 1 Preparation of Form A Crystals of 2-Hydroxy-5-[2-[4-[(2-pyridinylamino)sulfonyl]phenyl]diazenyl]-benzoic Acid D(-)-N-Methylglucamine Salt Sulfasalazine (2.00 g, 5.0 mmol) and D(-)-N-methylglucamine (1.00 g, 5.1 mmol) were weighed into a 250 ml round bottom flask equipped with a magnetic stirrer. Acetone (200 ml) was added and the mixture was stirred at 60°C. The solid material gradually dissolved and after a few hours a new precipitate started to form. The mixture never completely dissolved. After 24 hours at 60°C, tert-butyl methyl ether (40 ml) was added through the dropping funnel (5 min) and seed (1 mg of meglumine sulfasalazine Form A salt obtained as described in Example 5) was added. After 30 min, the heat was removed and the mixture was stirred at ambient temperature for a further 60 hours. It was then filtered (Robu-Glas borosilicate glass filter porosity 3) and the solid was washed with a 20% mixture of tert-butyl methyl ether in acetone (50 ml). The material was dried in vacuum for 17 hours and weighed on the filter to give 2.92 g (97.4%) of a yellow crystalline powder which was analyzed by 1H-NMR and found to contain 0.53% w / w acetone and traces of tert-butyl methyl ether (<0.02% w / w).

[0101] 1H NMR (400 MHz, DMSO-d6) δ 8.27 (d, J = 2.7 Hz, 1H), 8.03 - 7.95 (m, 3H), 7.91 - 7.83 (m, 2H), 7.80 (dd, J = 8.9, 2.7 Hz, 1H), 7.75 (ddd, J = 8.9, 7.1, 1.9 Hz, 1H), 7.21 (d, J = 8.7 Hz, 1H), 6.86 (t, J = 6.6 Hz, 1H), 6.73 (d, J = 8.9 Hz, 1H), 5.38 (s, 1H), 4.57 (s, 1H), 4.43 (s, 1H), 3.89 - 3.80 (m, 1H), 3.66 (dd, J = 5.3, 1.6 Hz, 1H), 3.60 (dd, J = 10.8, 3.2 Hz, 1H), 3.49 (dt, J = 8.9, 4.2 Hz, 1H), 3.45 - 3.37 (m, 2H), 3.05 (dd, J = 12.6, 3.3 Hz, 1H), 2.94 (dd, J = 12.6, 9.5 Hz, 1H), 2.55 (s, 3H); 13 C NMR (101 MHz, DMSO-d6) δ 170.96, 170.29, 154.12, 141.99, 127.70, 126.94, 126.78, 122.02, 119.07, 118.24, 71.28, 70.39, 70.10, 68.34, 63.27, 50.80; loss on drying (TGA; %w / w) is 0.2; melting point (DSC) is 163.5°C ± 2.5°C (onset); water vapor uptake (GVS; %w / w) is <0.4 at 30% RH and <0.9 at 80% RH; solubility in deionized water at 24°C and pH 6.6 is >54 mg / mL; NMR confirmed a 1:1 base to acid stoichiometry.

[0102] The powder X-ray diffraction pattern of crystalline Form A of the meglumine salt of sulfasalazine shown in FIG. 1 specifically includes the following XRPD peaks (expressed in degrees 2θ±0.2°): (10) 6.35, 13.93, and 22.41, or (11) 9.31, 15.86 and 20.99, or (12) 6.35, 13.93, 15.48, 15.86, 22.41, and 23.60; or (13) 6.35, 10.79, 12.93, 13.93, 15.48, 15.86, 18.12, 19.82, and 22.41; or (14) 9.31, 10.79, 12.93, 13.93, 14.47, 15.48, 15.86, 17.56, 19.10, 23.60, and 28.07; or (15) 6.35, 12.93, 13.93, 14.47, 15.48, 15.86, 19.10, 19.82, 20.99, 21.27, 22.41, 23.60, 23.89, and 28.07; or (16) 6.35, 9.31, 10.79, 12.93, 13.93, 14.47, 15.48, 15.86, 17.78, 18.12, 19.82, 20.11, 20.99, 21.27, 22.41, 23.60, 23.89, 28.07, and 28.80; or (17) 6.35, 9.31, 10.79, 12.93, 13.93, 14.47, 15.48, 15.86, 17.56, 17.78, 18.12, 18.50, 19.10, 19.82, 20.11, 20.99, 21.27, 22.41, 23.60, 23.89, 24.70, 25.14, 25.55, 25.93, 26.81, 28.07, and 28.80; or (18) 6.35, 9.31, 10.79, 12.93, 13.93, 14.47, 15.48, 15.86, 17.56, 17.78, 18.12, 18.50, 19.10, 19.82, 20.11, 20.99, 21.27, 22.41, 23.60, 23.89, 24.70, 25.14, 25.55, 25.93, 26.81, 28.07, 28.80, 29.49, 32.01, 32.58, 33.23.

[0103] Example 2 Preparation of Form A Crystals of 2-Hydroxy-5-[2-[4-[(2-pyridinylamino)sulfonyl]phenyl]diazenyl]-benzoic Acid D(-)-N-Methylglucamine Salt of the Present Invention To a suspension of 30 mg of sulfasalazine in acetone was added an equimolar amount of D(-)-N-methylglucamine (1 M stock solution in water). The suspension was heated to 23°C and the resulting solution was stirred for 4 days, after which the solvent was slowly evaporated. The salt product was washed, filtered and dried to give a polymorph of 2-hydroxy-5-[2-[4-[(2-pyridinylamino)sulfonyl]phenyl]diazenyl]-benzoic acid D(-)-N-methylglucamine salt designated as Form A.

[0104] 1 H-NMR (400 MHz, DMSO-d6) δ8.27 (d, J = 2.7 Hz, 1H), 8.05-7.95 (m, 3H), 7.91-7.83 (m, 2H), 7.80 (dd, J = 8.8, 2.7 Hz, 1H), 7.75 (ddd, J = 8.9, 7.2, 1.9 Hz, 1H), 7.21 (d, J = 8.7 Hz, 1H), 6.86 (t, J = 6.6 Hz, 1H), 6.73 (d, J = 8.8 Hz, 1H), 5.37 (s, 1H), 4.57 (s, 1H), 4.44 (s, 1H), 3.88-3.80 (m, 1H), 3.66 (dd, J = 5.2, 1.6 Hz, 1H), 3.60 (d, J = 10.7 Hz, 1H), 3.53-3.37 (m, 3H), 3.05 (dd, J = 12.6, 3.4 Hz, 1H), 2.94 (dd, J = 12.6, 9.5 Hz, 1H), 2.56 (s, 3H); 13 C-NMR (101 MHz, DMSO-d6) δ 171.01, 170.25, 154.13, 141.97, 127.70, 126.93, 126.78, 122.01, 119.06, 118.25, 71.28, 70.38, 70.09, 68.31, 63.26, 50.78, 40.12, 33.04; melting point (DSC): 160 °C ± 2 °C (onset); water vapor uptake (GVS; %w / w) was 0.4 at 30% RH and 1.1 at 80% RH; NMR and HPLC confirmed a 1:1 base to acid stoichiometry.

[0105] The powder X-ray diffraction pattern of crystalline Form A of the meglumine salt of sulfasalazine shown in FIG. 1 specifically includes the following XRPD peaks (expressed in degrees 2θ±0.2°): (1) 6.35, 13.93, and 22.41, or (2) 9.31, 15.86 and 20.99, or (3) 6.35, 13.93, 15.48, 15.86, 22.41, and 23.60; or (4) 6.35, 10.79, 12.93, 13.93, 15.48, 15.86, 18.12, 19.82, and 22.41; or (5) 9.31, 10.79, 12.93, 13.93, 14.47, 15.48, 15.86, 17.56, 19.10, 23.60, and 28.07; or (6) 6.35, 12.93, 13.93, 14.47, 15.48, 15.86, 19.10, 19.82, 20.99, 21.27, 22.41, 23.60, 23.89, and 28.07; or (7) 6.35, 9.31, 10.79, 12.93, 13.93, 14.47, 15.48, 15.86, 17.78, 18.12, 19.82, 20.11, 20.99, 21.27, 22.41, 23.60, 23.89, 28.07, and 28.80; or (8) 6.35, 9.31, 10.79, 12.93, 13.93, 14.47, 15.48, 15.86, 17.56, 17.78, 18.12, 18.50, 19.10, 19.82, 20.11, 20.99, 21.27, 22.41, 23.60, 23.89, 24.70, 25.14, 25.55, 25.93, 26.81, 28.07, and 28.80; or (9) 6.35, 9.31, 10.79, 12.93, 13.93, 14.47, 15.48, 15.86, 17.56, 17.78, 18.12, 18.50, 19.10, 19.82, 20.11, 20.99, 21.27, 22.41, 23.60, 23.89, 24.70, 25.14, 25.55, 25.93, 26.81, 28.07, 28.80, 29.49, 32.01, 32.58, 33.23.

[0106] Example 3 Preparation of Form A Crystals of 2-Hydroxy-5-[2-[4-[(2-pyridinylamino)sulfonyl]phenyl]diazenyl]-benzoic acid piperazine salt of the present invention To a suspension of 30 mg of sulfasalazine in methanol was added an equimolar amount of piperazine (1 M stock solution in water). The suspension was heated to 23° C. and stirred for 4 days. The salt product was then filtered, washed, filtered again and dried to obtain a polymorph designated piperazine sulfasalazine Form A.

[0107] 1 H-NMR (400 MHz, DMSO-d6) δ 8.28 (d, J = 2.7 Hz, 1H), 8.00 (dt, J = 6.9, 2.1 Hz, 3H), 7.88 (d, J = 8.6 Hz, 2H), 7.84 (dd, J = 8.8, 2.7 Hz, 1H), 7.75 (ddd, J = 8.9, 7.2, 1.9 Hz, 1 H), 7.21 (d, J = 8.7 Hz, 1 H), 6.86 (t, J = 6.4 Hz, 1H), 6.79 (d, J = 8.8 Hz, 1H), 3.18 (s, 8 H); 13C-NMR (101 MHz, DMSO-d6) δ 170.55, 169.77, 154.01, 142.36, 127.71, 127.22, 126.65, 122.10, 118.49, 118.22, 41.14; melting point (DSC): 270 °C ± 2 °C (heating rate 100 K / min); water vapor uptake (GVS; %w / w) was 0.4 at 30% RH and 0.9 at 80% RH; solubility in deionized water at 24 °C after 24 h: 0.12 mg / mL; NMR and HPLC confirmed a 1:2 base to acid stoichiometry.

[0108] The powder X-ray diffraction pattern of crystalline Form A of the piperazine salt of sulfasalazine, shown in FIG. 2, specifically includes the following XRPD peaks (expressed in degrees 2θ±0.2°): (1) 11.95, 12.30 and 16.42, or (2) 12.30, 12.93 and 15.00, or (3) 11.95, 12.30, 12.93, 16.42, 17.87, and 20.36; or (4) 8.11, 11.95, 12.30, 15.01, 16.42, 17.87, 20.36, and 20.74; or (5) 11.95, 12.30, 12.93, 15.01, 16.42, 17.87, 20.36, 20.74, 22.41, and 23.41; or (6) 11.95, 15.01, 16.42, 17.87, 20.36, 20.74, 23.41, 24.01, 24.67, 24.99, and 26.09; or (7) 8.11, 11.95, 12.30, 12.93, 15.01, 16.42, 17.87, 20.36, 20.74, 22.41, 23.41, 24.01, 24.67, 24.99, and 26.09, or (8) 11.95, 12.30, 12.93, 15.01, 16.42, 17.87, 20.36, 20.74, 22.41, 23.41, 24.01, 24.67, 24.99, 26.09, 26.81, 27.73, and 28.80; or (9) 8.11, 11.95, 12.30, 12.93, 15.01, 16.42, 17.87, 20.36, 20.74, 22.41, 23.41, 24.01, 24.67, 24.99, 26.09, 26.81, 27.73, 28.80, 29.80 and 30.43.

[0109] Example 4 Preparation of Form A crystals of 2-hydroxy-5-[2-[4-[(2-pyridinylamino)sulfonyl]phenyl]diazenyl]-benzoic acid diethylamine salt not covered by the present invention To a suspension of 30 mg of sulfasalazine in acetonitrile was added an equimolar amount of diethylamine (1 M stock solution in water). The suspension was heated to 23° C. and stirred for 4 days. The salt product was then filtered, washed, filtered again and dried to give a polymorph designated diethylamine sulfasalazine Form A.

[0110] 1 H-NMR (400 MHz, DMSO-d6) δ 8.26 (d, J = 2.7 Hz, 1H), 8.02-7.95 (m, 3H), 7.87 (d, J = 8.3 Hz, 2H), 7.79 (dd, J = 8.9, 2.7 Hz, 1H), 7.75 (t, J = 8.7, 7.6 Hz, 1 H), 7.21 (d, J = 8.6 Hz, 1 H), 6.86 (s, 1H), 6.72 (d, J = 8.8 Hz, 1H), 2.93 (q, J = 7.3 Hz, 4 H), 1.16 (t, J = 7.3 Hz, 6 H); 13 C-NMR (101 MHz, DMSO-d6) δ 170.21, 141.91, 126.91, 126.79, 121.99, 119.06, 118.27, 41.35, 11.05; melting point (DSC): 200 °C ± 2 °C (heating rate 10 K / min); water vapor uptake (GVS; %w / w) was 0.3 at 30% RH and 0.5 at 80% RH; solubility in deionized water at 24 °C after 24 h: >0.31 mg / mL; NMR and HPLC confirmed a 1:1 base to acid stoichiometry.

[0111] The powder X-ray diffraction pattern for crystalline Form A of the diethylamine salt of sulfasalazine, shown in FIG. 3, specifically includes the following XRPD peaks (expressed in degrees 2θ±0.2°): (1) 7.16, 11.48, and 18.78, or (2) 10.50, 15.41, and 21.87; or (3) 7.16, 10.50, 11.48, 18.78, 21.65, and 21.87; or (4) 10.50, 11.48, 12.42, 14.38, 15.41, 16.64, 18.78, and 21.87; or (5) 7.16, 10.50, 11.01, 11.48, 13.87, 15.92, 16.64, 18.78, 21.08, 21.65, and 22.15; or (6) 7.16, 10.50, 11.01, 11.48, 12.42, 13.87, 14.38, 15.41, 15.92, 16.64, 17.19, 18.28, 18.78, 21.08, 21.65, 21.87, and 22.15; or (7) 7.16, 10.50, 11.48, 12.42, 13.87, 14.38, 15.41, 15.92, 16.64, 20.52, 21.08, 21.65, 21.87, 22.15, 22.47, 23.16, 23.63, 24.14, and 25.11, or (8) 7.16, 10.50, 11.01, 11.48, 12.42, 13.87, 14.38, 15.41, 15.92, 16.64, 17.19, 18.28, 18.78, 20.52, 21.08, 21.65, 21.87, 22.15, 22.47, 23.16, 23.63, 24.14, and 25.11, or (9) 7.16, 10.50, 11.01, 11.48, 12.42, 13.87, 14.38, 15.41, 15.92, 16.64, 17.19, 18.28, 18.78, 20.52, 21.08, 21.65, 21.87, 22.15, 22.47, 23.16, 23.63, 24.14, 25.11, 26.94, 27.95, 28.92, 29.46.

[0112] Example 5 Preparation of Form B Crystals of Diethylamine 2-Hydroxy-5-[2-[4-[(2-pyridinylamino)sulfonyl]phenyl]diazenyl]-benzoic Acid Diethylamine Salt of the Present Invention 25 mg of sulfasalazine was added to 5 ml of acetone and a solution containing 10% excess of the counterion diethylamine in acetone was added. The mixture was heated to 45° C. and the solvent was slowly evaporated to give a polymorph designated diethylamine sulfasalazine salt Form B.

[0113] The powder X-ray diffraction pattern of crystalline Form B of the diethylamine salt of sulfasalazine, shown in FIG. 4, specifically includes the following XRPD peaks (expressed in degrees 2θ±0.2°): (1) 6.85, 17.82, and 22.75, or (2) 11.38, 20.58, and 23.98, or (3) 6.85, 11.38, 17.82, 20.58, and 22.75; or (4) 6.85, 11.38, 11.70, 17.62, 20.58, 22.75, and 23.98; or (5) 11.38, 11.70, 15.29, 16.71, 17.62, 19.92, 20.58, 21.30, 22.75, 23.63, and 23.98; or (6) 6.85, 11.38, 11.70, 14.78, 15.29, 15.70, 16.71, 17.62, 19.92, 20.20, 20.58, 21.30, 22.75, 23.63, 23.98, and 28.61; or (7) 6.85, 11.38, 11.70, 14.78, 15.29, 15.70, 16.71, 17.62, 19.92, 20.20, 20.58, 21.30, 22.75, 23.63, 23.98, 25.05, 25.71, 26.81, 27.95, and 28.61; or (8) 6.85, 11.38, 11.70, 14.78, 15.29, 15.70, 16.71, 17.62, 19.92, 20.20, 20.58, 21.30, 22.75, 23.63, 23.98, 25.05, 25.71, 26.81, 27.95, 28.61, 29.14, 31.06.

[0114] Example 6 Comparison of the transport rates of sulfasalazine free acid form and sulfasalazine meglumine Form A crystals across Caco-2 cell monolayers For transport experiments, Caco-2 cells were cultured at 1 cm 2 Transwell at a density of 67,800 cells per TM Cells were seeded onto filter inserts, which were placed into 12-well flat-bottom cluster plates. 0.5 mL of DMEM culture medium was placed in the inserts (apical compartment) and 1.5 mL in the outside wells (basal compartment). Cells were cultured in DMEM culture medium at 37°C, 10% CO2 and 90% relative humidity for 14-30 days until they formed a confluent monolayer. EVOM with STX-2 electrodes TM Confluency and adhesion of the cell monolayer were confirmed by measuring the transepithelial electrical resistance using a voltmeter. A TEER of 200 Ω×cm was obtained after preincubation (30 min) or after completion of the transport study. 2 If the efflux ratio was less than 0.05, the monolayer was rejected. Test items were made according to Biopharmaceutics Classification System (BCS) guidelines. Experiments were performed in triplicate. Immediately before the transport experiment, the cells were washed twice with Krebs-Ringer, after which the buffer was replaced with the transport solution. After 30 min preincubation, samples were drawn from both the donor and acceptor compartments. Six samples were taken at t = 0, 30, 60, 90, 120 and 180 min. The efflux ratio was calculated as P app (ba) / P app (ab), where P app (ba) is the apparent permeability coefficient for transport of the test compound from the basolateral to the apical side (secretory direction), P app(ab) is the apparent permeability coefficient for transport of the test compound from the apical side to the basolateral side (absorption direction). The apparent permeability coefficient P app (cm / s) was calculated as the permeability at steady state (μg / s) × (1 / initial amount of test compound in the donor compartment (μg) × 1 / area of ​​exposed cell monolayer (cm 2 ) × volume of buffer in donor compartment (cm 3 ).

[0115] [Table 1] Table 1 shows that greater amounts of sulfasalazine meglumine salt Form A crystals were cumulatively transported through the Caco-2 cell monolayer. The apparent permeability of sulfasalazine and meglumine salt Form A ranged from 1.5 (50 μg / mL) to 3.6 (200 μg / mL).

[0116] Example 7 Comparison of the pharmacokinetics of sulfasalazine after intravenous or oral administration in the free acid form or as crystalline Form A of the meglumine salt.

[0117] List of abbreviations AUCinf.: area under the curve to infinity; AUClast.: area under the curve to the last data point; Cmax: maximum concentration; CMC: cyclic methylcellulose; F: bioavailability; IV: intravenous; LC: liquid chromatography; MS: mass spectrometry; NCA: noncompartmental analysis; PK: pharmacokinetic; PO: oral; SSZ: sulfasalazine; TI: test item; Tmax: time of maximum concentration; T 1 / 2 :Half-life

[0118] In vivo test procedure In accordance with Swedish law and approval of the experimental procedures by the local ethical committee (M388-12) for preclinical in vivo testing and subsequent evaluation in rodents, after a minimum of 7 days of habituation to the housing environment after arrival, male Wistar (Hannover) rats (Taconic, Denmark), average body weight 180-200 g; average age 8-10 weeks, were treated with the test items (see Table 2); 12-16 hours before administration, all food was removed except for an amount equal to half-day consumption. The test items were administered using a soft feeding tube (po) or the test items were injected into the tail vein. The volume administered was 10 mL / kg (po) or 1 mL / kg (iv). Before iv administration, the rats were anesthetized with isoflurane. During sample collection, the rats were conscious and blood was collected from the sublingual vein. Blood samples were collected from each rat over a period of up to 24 hours. At each time point, two aliquots of 50 μL were added to vials each containing 150 μL of sterile water. Samples were mixed immediately and stored at −18° C. until preparation for bioanalysis began. All formulations were prepared on the same day that dosing occurred. The rats' body weights were recorded prior to dosing. The weights of the syringes were recorded before and after dosing, and the exact amount of test sample delivered was calculated. The exact dose was used during data evaluation. [Table 2]

[0119] Bioanalysis Plasma levels of sulfasalazine were determined using LCMS / MS in mrm (multiple reaction monitoring) mode.

[0120] Samples and standards were injected by HTC PAL from a Shimadzu LC system CTC analytics consisting of a high pressure gradient system of two LC-10 AD pumps controlled by a Shimadzu SCL-10A controller. Samples were separated using reversed phase chromatography with gradient elution at a flow rate of 0.8 mL / min. The mobile phases were A 94.9 / 5 / 0.1 water / acetonitrile / formic acid and B 5 / 94.9 / 0.1 water / acetonitrile / formic acid. The gradient started from 0% B and increased linearly to 100% B in 4 min, maintained at 100% B for 0.5 min, then the system was returned to 0% B in 0.1 min. The system was equilibrated for 1.4 min until the total run time was 6 min. The eluate was analyzed by a Micromass Quattro Ultima equipped with an electrospray ion source. Data was collected and calibration was calculated by MassLynx 4.0 software. Sulfasalazine was separated on a Waters Symmetry C18 50 × 2.1 column. The eluate was ionized by negative ion electrospray and the mrm transition from 397 to 197 m / z was monitored.

[0121] Diluted blood samples (50 μL blood, 150 μL water) were thawed and mixed. To precipitate proteins, 400 μL acetonitrile was added and mixed again. Samples were centrifuged at 5,000 g for 5 min. 100 μL of the supernatant was transferred to a 300 μL glass vial and 100 μL water was added to reduce the acetonitrile concentration.

[0122] Positive and negative mode MS / MS were used for sulfasalazine. Standard curve concentrations ranged from 5 nM to 15625 nM. Samples with analyte concentrations above the upper limit of quantification were diluted with matrix to reach within the assay range. Noncompartmental analysis (NCA) was performed using the Phoenix WinNonLin analysis tool.

[0123] result Intravenous The pharmacokinetic profile of sulfasalazine following iv administration of 1 and 5 mg / kg sulfasalazine meglumine salt Form A is summarized in Table 3. There was good linearity between exposure at the two doses, with estimated half-lives of 1.2 and 1.3 hours, respectively, which is in good agreement with published data (Zamek-Gliszczynski MJ et al. Characterization of SAGE Mdr1a (P-gp), Bcrp, and Mrp2 knockout rats using loperamide, paclitaxel, sulfasalazine, and carboxydichloroflurorescein pharmacokinetics. Drug Metab Dispos 2012, 40, 1825). [Table 3]

[0124] Oral route The pharmacokinetic profile of sulfasalazine after oral administration of 60 mg / kg is summarized in Table 4. The maximum concentration of sulfasalazine was increased 7-fold for 2-hydroxy-5-[2-[4-[(2-pyridinylamino)sulfonyl]phenyl]diazenyl]-benzoic acid D(-)-N-methylglucamine salt form A compared with sulfasalazine and reached 6-fold faster for 2-hydroxy-5-[2-[4-[(2-pyridinylamino)sulfonyl]phenyl]diazenyl]-benzoic acid D(-)-N-methylglucamine salt form A compared with sulfasalazine. Total plasma levels of sulfasalazine were >50% (AUC inf ) and increased by >90% during the first 90 minutes after oral administration of the meglumine salt. The bioavailability of sulfasalazine meglumine Form A in rats was increased by approximately 50% over the bioavailability of the free acid form of sulfasalazine. [Table 4]

[0125] Example 8 Comparison of the dissolution rates of sulfasalazine free acid form and crystalline form A of the meglumine salt of sulfasalazine.

[0126] HPLC The amount of dissolved sulfasalazine was determined by HPLC on a HP 1100 instrument using a Waters X-Bridge 3.5 μm C18 column (150 × 4.6 mm) and a gradient method, respectively. The mobile phase consisted of the following: Mobile phase (A): 1.13 g of sodium dihydrogen phosphate and 2.5 g of sodium acetate were dissolved in 1000 mL of purified water. The pH was then adjusted to 4.8 with acetic acid (100%). Mobile phase (B): 1 part of mobile phase A is mixed with 4 parts of chromatographic grade methanol. The flow rate was 1.0 mL / min, the injection volume was 5 μL, and the detection wavelength was 320 nm (HP DAD series 1100). The run time was 10 min. Quantification was performed using an external standard method. The quantitative method was validated for selectivity, reproducibility, and linearity. Samples are prepared in diluted ammonia R3 PhEur.

[0127] 50 mg of sulfasalazine and meglumine salt form A of sulfasalazine were weighed into 4 mL glass vials containing 2 mL of water or FaSSIF-V2 medium, respectively, and stirred at room temperature (20-25°C) for 24 h, after which 20 mg of sulfasalazine and meglumine salt form A of sulfasalazine were added until a saturated solution was obtained. The saturated solution was filtered using a centrifugal filter (Nylon, 0.45 μM) and the clear supernatant was injected directly or after dilution with diluted ammonia PhEur, respectively. [Table 5]

Claims

1. A method for preparing crystals of an organic salt of 2-hydroxy-5-[2-[4-[(2-pyridinylamino)sulfonyl]phenyl]diazenyl]-benzoic acid (sulfasalazine), wherein the organic salt is selected from diethylamine, piperazine and D-(-)-N-methylglucamine (meglumine), the method comprising the steps of: A1: Providing sulfasalazine free acid form in a suitable solvent; B1: Providing an organic amine-containing component selected from meglumine and piperazine in a suitable solvent; C1: mixing the sulfasalazine solution of step A1) with the organic amine-containing component solution of step B1) at room temperature, preferably 19°C to 25°C; and D1: isolating the crystals of piperazine sulfasalazine Form A or a solvate thereof or the crystals of meglumine sulfasalazine Form A or a solvate thereof formed in the solution of step C1); or A2: Providing sulfasalazine free acid form in a suitable solvent; B2: Providing an organic amine-containing component selected from meglumine and diethylamine in a suitable solvent; C2: mixing the sulfasalazine solution of step A2) with the organic amine-containing component solution of step B2), wherein the amine-containing component is in at least a 5% molar excess relative to the sulfasalazine, more preferably at least a 10% molar excess, more preferably at least a 20% molar excess; and D2a: Concentrating the mixture formed in step C2) to separate crystals of meglumine sulfasalazine Form A or a solvate thereof or crystals of diethylamine sulfasalazine Form B or a solvate thereof; or D2b: adding an additional solvent to the mixture formed in step C2), wherein the additional solvent is different from the solvent used in steps A2) and B2), and isolating the crystals of meglumine sulfasalazine Form A or a solvate thereof or the crystals of diethylamine sulfasalazine Form B or a solvate thereof formed in step D2b).

23. A method of manufacturing comprising or consisting of:

2. the solvent of step A1) is selected from suitable organic solvents, preferably selected from the group consisting of acetone, acetonitrile, and alcohols, such as methanol, ethanol, propanol, isopropanol and butanol, and / or - the solvent in step B1) is water or comprises water, and / or - in step C1) using equimolar amounts of sulfasalazine and the organic amine-containing component, and / or - in step C1), mixing the solution of sulfasalazine and the organic amine-containing component for at least 24 hours at room temperature, preferably at least 48 hours at room temperature, more preferably at least 72 hours at room temperature, even more preferably at least 96 hours at room temperature; The method according to claim 1, characterized in that

3. the solvent for steps A2) and B2) is selected from suitable organic solvents, preferably selected from the group consisting of acetone, acetonitrile and alcohols, such as methanol, ethanol, propanol, isopropanol and butanol, more preferably the solvent for steps A2) and B2) is acetone, and / or - in step C2) mixing the solutions of sulfasalazine and the organic amine-containing component at elevated temperature, preferably at a temperature in the range of from 30°C to 80°C, more preferably from 40°C to 65°C; and / or - in step C2), mixing the solution of sulfasalazine and the organic amine-containing component for up to 24 hours; and / or - in step D2b), mixing the solution containing the further solvent for at least 24 hours at room temperature, preferably at a temperature ranging from 19°C to 25°C, preferably for at least 48 hours at room temperature, more preferably for at least 60 hours at room temperature. The method according to claim 1, characterized in that

4. 3. The process according to claim 1 or 2, wherein the produced piperazine sulfasalazine Form A crystals are characterized by X-ray powder diffraction peaks at 2θ values ​​(±0.2) of 12.30, 12.93, 15.01, 16.42, 22.41 and 23.4, preferably having an X-ray powder diffraction pattern substantially identical to that shown in Figure 2.

5. The process according to any one of claims 1 to 3, wherein the produced meglumines sulfasalazine Form A crystals are characterized by X-ray powder diffraction peaks at 2θ values ​​(±0.2) of 6.35, 13.93, 15.48, 15.86, 20.99, 22.41, 23.60 and 28.07, preferably having an X-ray powder diffraction pattern substantially identical to that shown in Figure 1.

6. The method according to any one of claims 1 to 3, characterized in that in step D1), the prepared crystals of piperazine sulfasalazine Form A or a solvate thereof or the prepared crystals of meglumine sulfasalazine Form A or a solvate thereof are treated by a suitable method to form anhydrous crystals, or in step D2a) or D2b), the prepared crystals of diethylamine sulfasalazine Form B or a solvate thereof or the prepared crystals of meglumine sulfasalazine Form A or a solvate thereof are treated by a suitable method to form anhydrous crystals.

7. 4. The process according to claim 1 or 3, wherein the produced diethylamine sulfasalazine Form B crystals are characterized by X-ray powder diffraction peaks at 2θ values ​​of 6.85, 11.38, 11.70, 17.62, 20.58, 22.75 and 23.98 (±0.2), preferably having an X-ray powder diffraction pattern substantially identical to that shown in Figure 4.

8. crystalline meglumine sulfasalazine Form A obtained according to any one of claims 1 to 3, 5 and 6; or crystalline piperazine sulfasalazine Form A obtained according to any one of claims 1, 2, 4 or 6; or crystalline diethylamine sulfasalazine Form B obtained according to any one of claims 1, 3, 6 and 7.

9. 10. A pharmaceutical composition comprising a therapeutically effective amount of one or more of the crystalline salt forms of sulfasalazine of claim 8.

10. 10. The pharmaceutical composition according to claim 9, characterized in that the composition is for oral or rectal administration.

11. The pharmaceutical composition preferably comprises a non-steroidal anti-inflammatory drug; preferably a non-selective cyclo-oxygenase COX-1 / COX-2 inhibitor, whether applied topically or systemically, such as piroxicam, diclofenac, propionic acids, such as naproxen, flurbiprofen, fenoprofen, ketoprofen and ibuprofen, fenamates, such as mefenamic acid, indomethacin, sulindac, azapropazones, pyrazolones, such as phenylbutazone, salicylates, such as aspirin, selective COX-2 inhibitors, such as meloxicam, celecoxib, rofecoxib, valdecoxib, lumiracoxib, parecoxib and etoricoxib, nitric oxide donating cyclooxygenase inhibitors, such as ... Oxygenase inhibitors (CINOD); glucocorticoids, preferably flunisolide, triamcinolone acetonide, betamethasone dipropionate, budesonide, fluticasone propionate, ciclesonide or mometasone furoate; methotrexate; leflunomide; hydroxychloroquine; d-penicillamine; diacerein; dietary supplements, preferably glucosamine; gold, preferably auranofin; cytokines or agonists or antagonists of cytokine function; monoclonal antibodies targeting B lymphocytes, preferably CD20 (rituximab); MRA-aIL16R; T-lymphocytes; CTLA4-Ig; HuMax 11-15; modulators of chemokine receptor function, preferably antagonists of CCR2, CCR2A, CCR2B, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCRlO and CCR11 (CC family), CXCRl, CXCR2, CXCR3, CXCR4 and CXCR5 (CXC family) and CX3CR1 (C-X3-C family); azathioprine, tofacitinib, monoclonal antibodies such as the anti-tumor necrosis factor alpha monoclonal antibodies infliximab, adalimumab and golimumab; interleukin 1 receptor antagonists such as anakinra; etanercept and abatacept;11. A pharmaceutical composition according to claim 9 or 10, characterized in that it further comprises one, two, three or more active ingredients, more preferably selected from the group consisting of methotrexate and hydroxychloroquine.

12. i) diseases or conditions in which modulation of inflammatory cells is beneficial; ii) diseases or conditions relating to bones or joints, preferably arthritis associated with or including osteoarthritis, primary or secondary to, for example, congenital hip dysplasia; cervical and lumbar spondylitis, and lower back and neck pain; rheumatoid arthritis and Still's disease; seronegative spondyloarthropathy, such as ankylosing spondylitis, psoriatic arthritis, reactive arthritis and undifferentiated spondyloarthropathy, septic arthritis and other arthropathies and bone disorders associated with infections, such as tuberculosis, e.g. Pott's disease and Ponce's disease; acute and chronic crystal-induced synovitis, such as uric acid gout, calcium pyrophosphate deposition disease, and calcium apatite-associated tendon, bursa and synovial inflammation; Behcet's disease; primary or secondary Sjogren's syndrome; systemic sclerosis and localized forms. scleroderma; systemic lupus erythematosus, mixed connective tissue disease, and undifferentiated connective tissue disease; inflammatory myopathies, such as dermatomyositis and polymyositis; polymyalgia rheumatica; juvenile arthritis, such as idiopathic inflammatory arthritis of any joint location and associated syndrome, and rheumatic fever and its systemic complications; vasculitis, such as giant cell arteritis, Takayasu's arteritis, Churg-Strauss syndrome, polyarteritis nodosa, microscopic polyarteritis, and vasculitis associated with viral infections, hypersensitivity reactions, cryoglobulins, and paraproteins; lower back pain; familial Mediterranean fever, Muckle-Wells syndrome, and familial Irish fever, Kikuchi's disease; drug-induced arthralgia, tendonitis, and myopathy; and iii) diseases or conditions relating to the digestive tract, preferably selected from the group consisting of eosinophilic gastroenteritis, mastocytosis, Crohn's disease, colitis, e.g. ulcerative colitis, proctitis; celiac disease, irritable bowel syndrome, and food-related allergies that may affect areas distant from the intestine, e.g. migraine, rhinitis or eczema. A pharmaceutical composition according to any one of claims 9 to 11 for use in the treatment of

13. i) diseases or conditions in which modulation of inflammatory cells is beneficial; ii) diseases or conditions relating to bones or joints, preferably arthritis associated with or including osteoarthritis, primary or secondary to, for example, congenital hip dysplasia; cervical and lumbar spondylitis, and lower back and neck pain; rheumatoid arthritis and Still's disease; seronegative spondyloarthropathy, such as ankylosing spondylitis, psoriatic arthritis, reactive arthritis and undifferentiated spondyloarthropathy, septic arthritis and other arthropathies and bone disorders associated with infections, such as tuberculosis, e.g. Pott's disease and Ponce's disease; acute and chronic crystal-induced synovitis, such as uric acid gout, calcium pyrophosphate deposition disease, and calcium apatite-associated tendon, bursa and synovial inflammation; Behcet's disease; primary or secondary Sjogren's syndrome; systemic sclerosis and localized forms. scleroderma; systemic lupus erythematosus, mixed connective tissue disease, and undifferentiated connective tissue disease; inflammatory myopathies, such as dermatomyositis and polymyositis; polymyalgia rheumatica; juvenile arthritis, such as idiopathic inflammatory arthritis of any joint location and associated syndrome, and rheumatic fever and its systemic complications; vasculitis, such as giant cell arteritis, Takayasu's arteritis, Churg-Strauss syndrome, polyarteritis nodosa, microscopic polyarteritis, and vasculitis associated with viral infections, hypersensitivity reactions, cryoglobulins, and paraproteins; lower back pain; familial Mediterranean fever, Muckle-Wells syndrome, and familial Irish fever, Kikuchi's disease; drug-induced arthralgia, tendonitis, and myopathy; and iii) diseases or conditions relating to the digestive tract, preferably selected from the group consisting of eosinophilic gastroenteritis, mastocytosis, Crohn's disease, colitis, e.g. ulcerative colitis, proctitis; celiac disease, irritable bowel syndrome, and food-related allergies that may affect areas distant from the intestine, e.g. migraine, rhinitis or eczema. Use of a crystalline salt form of sulfasalazine according to claim 8 or a pharmaceutical composition according to any one of claims 9 to 11 in the manufacture of a medicament for / in the treatment of.

14. i) diseases or conditions in which modulation of inflammatory cells is beneficial; ii) diseases or conditions relating to bones or joints, arthritis, primary or associated with or including osteoarthropathy secondary to, for example, congenital hip dysplasia; cervical and lumbar spondylitis, and lower back and neck pain; rheumatoid arthritis and Still's disease; seronegative spondyloarthropathy, e.g. ankylosing spondylitis, psoriatic arthritis, reactive arthritis and undifferentiated spondyloarthropathy, septic arthritis and other arthropathies and bone disorders associated with infection, e.g. tuberculosis, e.g. Pott's disease and Ponce's disease; acute and chronic crystal-induced synovitis, e.g. uric acid gout, calcium pyrophosphate deposition disease, and calcium apatite-associated tendon, bursa and synovial inflammation; Behcet's disease; primary or secondary Sjogren's syndrome; systemic sclerosis and localized scleroderma. systemic lupus erythematosus, mixed connective tissue disease, and undifferentiated connective tissue disease; inflammatory myopathies, such as dermatomyositis and polymyositis; polymyalgia rheumatica; juvenile arthritis, such as idiopathic inflammatory arthritis of any joint location and associated syndrome, and rheumatic fever and its systemic complications; vasculitis, such as giant cell arteritis, Takayasu's arteritis, Churg-Strauss syndrome, polyarteritis nodosa, microscopic polyarteritis, and vasculitis associated with viral infections, hypersensitivity reactions, cryoglobulins, and paraproteins; lower back pain; familial Mediterranean fever, Muckle-Wells syndrome, and familial Irish fever, Kikuchi's disease; drug-induced arthralgia, tendonitis, and myopathy; and iii) diseases or conditions relating to the digestive tract, preferably selected from the group consisting of eosinophilic gastroenteritis, mastocytosis, Crohn's disease, colitis, e.g. ulcerative colitis, proctitis; celiac disease, irritable bowel syndrome, and food-related allergies that may affect areas distant from the intestine, e.g. migraine, rhinitis or eczema.

12. A method for treating a disease or condition in a patient suffering from or at risk of said disease or condition, comprising administering to the patient a therapeutically effective amount of a crystalline salt form of sulfasalazine according to claim 8 or a pharmaceutical composition according to any one of claims 9 to 11.

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