TW201710256A - New pharmaceutical salt forms and crystalline forms - Google Patents

Abstract

The present invention relates to new salt forms of (3S)-Tetrahydrofuran-3-yl(4S)-4-isopropyl-1,4,6,7-tetrahydro -5H-imidazo[4,5-c]pyridine-5-carboxylate, and to their use in medicine. This invention relates also to a crystalline mesylate salt form of (3S)-Tetrahydrofuran-3-yl(4S)-4-isopropyl-1,4,6, 7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylate, and to the use of the same in medicine.

Description

Novel pharmaceutical salt forms and crystalline forms

The present invention relates to (4S)-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3S)-tetrahydrofuran-3 a novel salt form of a base ester and its use in medicine. The invention also relates to (4S)-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3S)-tetrahydrofuran-3 - Crystalline mesylate form of the base ester, and its use in medicine.

The semicarbazide-sensitive amine oxidase (SSAO) activity is derived from Vascular Adhesion Protein-1 (VAP-1) or Amine Oxidase (Amine Oxidase). Copper Containing 3) (AOC3) (belonging to the enzyme activity of the copper amine oxidase enzyme family (EC. 1.4.3.6)). Therefore, inhibitors of SSAO enzymes can also regulate the biological function of VAP-1 protein.

SSAO activity has occurred in a variety of different tissues, including vascular and non-vascular smooth muscle tissue, endothelium, and adipose tissue [Lewinsohn, Braz. J. Med. Biol. Res. 1984 , 17 , 223-256; Nakos & Gossrau, Folia Histochem. Cytobiol. 1994 , 32 , 3-10; Yu et al., Biochem. Pharmacol. 1994 , 47 , 1055-1059; Castillo et al . , Neurochem. Int . 1998 , 33 , 415-423; Lyles & Pino, J. Neural. Transm. Suppl. 1998 , 52 , 239-250; Jaakkola et al . , Am. J. Pathol. 1999 , 155 , 1953-1965; Morin et al. J. Pharmacol. Exp. Ther. 2001 , 297 , 563-572; Salmi & Jalkanen, Trends Immunol. 2001 , 22 , 211-216]. In addition, SSAO proteins are present in plasma, and this soluble form appears to have properties similar to the tissue-bound morphology [Yu et al. , Biochem. Pharmacol. 1994 , 47 , 1055-1059; Kurkijärvi et al. , J. Immunol. 1998 , 161 , 1549-1557].

The correct physiological role of this variety of enzymes has not been fully established, but SSAO and its reaction products appear to have several functions in cell signal transduction and regulation. For example, recent results show that SSAO is GLUT4-mediated glucose uptake [Enrique-Tarancon et al. J. Biol. Chem. 1998 , 273 , 8025-8032; Morin et al. J. Pharmacol. Exp. Ther. 2001 , 297 , 563-572] plays a role in the differentiation of adipocytes [Fontana et al. , Biochem. J. 2001 , 356 , 769-777; Mercier et al. , Biochem. J. 2001 , 358 , 335-342] . In addition, SSAO has been shown to be involved in the inflammatory process as an adhesion protein for leukocytes [Salmi & Jalkanen, Trends Immunol. 2001 , 22 , 211-216; Salmi & Jalkanen in " Adhesion Molecules: Functions and Inhibition " K.Ley (Ed .), 2007 , pp. 237-251], and may also play a role in the development and maintenance of connective tissue matrix [Langford et al. Cardiovasc. Toxicol. 2002 , 2(2) , 141-150; Göktürk et al. Am. J. Pathol. 2003 , 163(5) , 1921-1928]. In addition, the correlation between SSAO and angiogenesis has recently been found [Noda et al., FASEE J. 2008 , 22(8) , 2928-2935], and based on this correlation, inhibitors of SSAO are expected to have an anti-angiogenic effect. .

Several human trials have confirmed that SSAO activity in plasma is elevated in conditions such as congestive heart failure, diabetes, Alzheimer's disease, inflammation, etc. [Lewinsohn, Braz. J. Med. Biol. Res. 1984 , 17 , 223-256; Boomsma et al . , Cardiovasc. Res . 1997 , 33 , 387-391; Ekblom, Pharmacol. Res . 1998 , 37 , 87-92; Kurkijärvi et al. J. Immunol. 1998 , 161 , 1549- 1557; Boomsma et al., Diabetologia 1999 , 42 , 233-237; Meszaros et al . , Eur. J. Drug Metab. Pharmacokinet. 1999 , 24 , 299-302; Yu et al., Biochim. Biophys. Acta 2003 , 1647 (1) -2) , 193-199; Mátyus et al. Curr. Med. Chem. 2004 , 11(10) , 1285-1298; Neurotoxicology 2004 , 25(1-2) , 303-315; del O'Sullivan et al. Mar Hernandez et al . , Neurosci. Lett. 2005 , 384(1-2) , 183-187]. Reactive aldehydes and hydrogen peroxide produced by endogenous amine oxidase have been shown to cause cardiovascular disease, diabetic complications and Alzheimer's disease [Callingham et al . Prog. Brain Res. 1995 , 106 , 305- 321; Ekblom, Pharmacol. Res . 1998 , 37 , 87-92; Yu et al., Biochim. Biophys. Acta 2003 , 1647 (1-2) , 193-199; Jiang et al., Neuropathol Appl Neurobiol. 2008 , 34 ( 2) , 194-204]. In addition, SSAO is known to be strongly expressed in the vascular endothelium, indicating that SSAO enzyme activity is involved in the process of leukocyte vascular exudation at the site of inflammation [Salmi et al. Immunity 2001 , 14(3) , 265-276; Salmi & Jalkanen described in " Adhesion Molecules: Functions and Inhibition " K. Ley (Ed.), 2007 , pp. 237-251]. Therefore, it has been proposed to inhibit SSAO, which has the medical value of preventing diabetic complications and inflammatory diseases [Ekblom, Pharmacol. Res . 1998 , 37 , 87-92; Salmi et al. Immunity 2001 , 14(3) , 265-276; Salter- Cid et al. , J. Pharmacol. Exp. Ther. 2005 , 315(2) , 553-562].

WO2007/146188 teaches that blocking SSAO activity can inhibit leukocyte recruitment, reduce inflammatory response, and is expected to be beneficial in preventing and treating, for example, epileptic seizures.

O'Rourke et al. (J Neural Transm. 2007; 114(6): 845-9) explored the potential of SSAO inhibitors in neurological diseases, as the effects of inhibiting SSAO have been demonstrated in rat stroke patterns in the past. SSAO inhibitors are tested in relapsing-remitting experimental autoimmune encephalomyelitis (EAE), a mouse model that exhibits many of the common features of human multiple sclerosis. The data confirms the potential clinical benefit of small molecule anti-SSAO therapy in this model and therefore treats human multiple sclerosis.

Animals that excluded SSAO were apparently normal in phenotype, but the inflammatory response to various inflammatory stimuli was significantly reduced [Stolen et al. Immunity 2005 , 22(1) , 105-115]. In addition, in a variety of animal models of human disease (eg, carrageenan-induced paw inflammation, In the presence of oxazolone-induced colitis, lipopolysaccharide-induced lung inflammation, collagen-induced arthritis, endotoxin-induced uveitis, the use of antibodies and/or small molecules to antagonize their function in wild-type animals has shown protection Sex, can reduce white blood cell infiltration, reduce the severity of disease phenotype, and reduce inflammatory cytokines and chemical content [Kirton et al. Eur. J. Immunol. 2005 , 35 (11) , 3119-3130; Salter-Cid, etc. J. Pharmacol. Exp. Ther. 2005 , 315(2) , 553-562; McDonald et al., Annual Reports in Medicinal Chemistry 2007 , 42 , 229-243; Salmi & Jalkanen, "Adhesion Molecules: Functions and Inhibition "K.Ley (Ed.), 2007, pp.237-251; Noda , who's FASEB J. 2008 22 (4), 1094-1103; Noda , who's FASEB J. 2008, 22 (8) , 2928- 2935]. This anti-inflammatory protective effect appears to have spanned a wide range of inflammatory patterns, which have their own independent mechanisms of action, rather than being limited to a particular disease or disease pattern. This point will show that SSAO may be a key point in regulating this inflammatory response, so SSAO inhibitors may be effective anti-inflammatory drugs for a variety of human diseases. VAP-1 has also been implicated in the deterioration and maintenance of fibrotic diseases, including fibrotic diseases of the liver and lungs. Weston and Adams (J Neural Transm. 2011 , 118(7), 1055-64) have summarized experimental data on VAP-1 in liver fibrosis, and Weston et al. (EASL Poster 2010) suggested that blocking VAP-1 can accelerate Analyze the fibrosis induced by carbon tetrachloride. In addition, VAP-1 has been involved in inflammation of the lungs (eg, Singh et al. 2003 , Virchows Arch 442: 491-495), suggesting that VAP-1 blockers can reduce lung inflammation and therefore can be used to treat disease-promoting fibrosis With pro-inflammatory and beneficial to treat cystic fibrosis.

SSAO (VAP-1) is up-regulated in gastric cancer and has been identified in human melanoma, hepatoma, and tumor vessels of head and neck tumors (Yoong KF, McNabG, Hubscher SG, Adams DH. (1998), J Immunol 160 , 3978-88.; Irjala H, Salmi M, Alanen K, Gre'nman R, Jalkanen S (2001), Immunol. 166 , 6937-6943; Forster-Horvath C, Dome B, Paku S et al ( 2004), Melanoma Res. 14 , 135-40.). A report (Marttila-Ichihara F, Castermans K, Auvinen K, Oude Egbrink MG, Jalkanen S, Griffioen AW, Salmi M. (2010), J Immunol. 184 , 3164-3173.) has been shown to have an enzyme-inert VAP- The melanoma of mice in 1 has a slower growth, and the number and diameter of tumor blood vessels have decreased. The decrease in growth of these tumors also reflects a decrease in infiltration of myelosuppressive cells (60-70% reduction). Encouragingly, VAP-1 deficiency did not affect blood vessels or lymphoid formation in normal tissues.

For the above reasons, it is hoped that inhibition of SSAO will reduce the content of pro-inflammatory enzyme products (aldehydes, hydrogen peroxide and ammonia), as well as reduce the adhesion of immune cells and their corresponding activating and final exudation. Diseases that are expected to be medically beneficial as such include all diseases that are significantly affected by immune cells in the initiation, maintenance, or resolution of the disease, such as inflammatory diseases and immune/autoimmune diseases. Examples of such diseases include multiple sclerosis, arthritis, and vasculitis.

Novel and improved SSAO inhibitors are still unable to meet medical needs. WO2010 / 031789 (the disclosure of which is incorporated herein by reference in it) discloses a reliable SSAO inhibitor compounds, especially of Example 16, which system (4 S) -4- isopropyl -1,4,6, 7- tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran-3-yl ester free base of, has the following structural formula:

(4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl ester The free base is a hygroscopic amorphous glass/colloid. The glass transition point is 39 ° C at a relatively low temperature, so the free base is often colloidal.

The invention described herein relates to a system having improved properties of surprising SSAO inhibitor (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine 5-carboxylic acid (3 S) - tetrahydrofuran-3-yl novel form of a salt of the ester.

Hygroscopicity is an undesirable property of the drug because inhaling water causes many problems. Examples of such problems include that the amount of water changes so that the drug is difficult to weigh, and it is difficult to manipulate the drug because it is easily sticky. Gum is usually not desired because it is sticky and difficult to handle. Crystallization is better than amorphous gel, because crystals have better filtration properties and are therefore easier to dry.

It is also hoped that the drug will have good thermal stability. The temperature during the conventional grinding and tableting process generally exceeds 50 ° C (see, for example, Developing Solid Oral Dosage Forms: Pharmaceutical Theory &Practice; Yihong Qiu, Yisheng Chen, Geoff G. Z. Zhang, Lirong Liu, William Porter, 2009). A significant risk is that "hot spots" can occur during the grinding or tableting process, and the temperature of the hot spot will exceed the melting point of the drug. The molten drug that occurs during the grinding or tableting process can cause the drug particles to agglomerate or form agglomerates. Such melting, agglomeration or agglomeration will hinder accuracy and consistency.

Depth study (4 S) -4-isopropyl-l, 4,6,7-tetrahydro hereinafter -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran -3 - The preparation and properties of the base ester free base and its salt form, the Applicant has found an advantageous salt form, i.e., a methanesulfonate salt, which has the advantages of high thermal stability and favorable low hygroscopicity; and sulfate, It is a hydrate with favorable high heat stability and favorable low hygroscopicity.

The present invention includes one comprising a (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran A sulphate or methanesulfonate salt of a -3-yl ester, and a composition of one or more pharmaceutically acceptable excipients.

(4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl The sulfate and methanesulfonate esters are useful for treating inflammatory, inflammatory, immune or autoimmune disorders, or inhibiting tumor growth. In a specific embodiment, the inflammatory or inflammatory disease or immune or autoimmune disorder is arthritis (including rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis and dry arthritis), synovitis, Vasculitis, Sjogren's disease, conditions associated with inflammation of the intestines (including Crohn's disease, ulcerative colitis, inflammatory bowel disease and intestinal bowel disease), atherosclerosis, multiple Hardening, Alzheimer's disease, vascular dementia, Parkinson's disease, cerebral amyloid angiopathy, somatic cerebral arterial disease complicated with subcortical cerebral infarction and white matter lesions, pulmonary inflammatory disease ( Including asthma, chronic obstructive pulmonary disease and acute respiratory distress syndrome), fibrotic diseases (including idiopathic pulmonary fibrosis, cardiac fibrosis, liver fibrosis and systemic sclerosis (scleroderma)), skin inflammation (including contact dermatitis, atopic dermatitis and dryness), eye inflammation (including age-related macular degeneration, uveitis and diabetic retinopathy), systemic inflammatory response syndrome , sepsis, inflammation of the liver and / or autoimmune disorders (including autoimmune hepatitis, primary biliary cirrhosis, alcoholic liver disease, sclerosing cholangitis, and autoimmune cholangitis), diabetes (I Or type II) and/or its complications, chronic heart failure, congestive heart failure, ischemic disease (including stroke and ischemia-reperfusion injury) or myocardial infarction and/or its complications, or epilepsy.

The present invention includes the (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran - The use of a sulfate and a methanesulfonate salt of a 3-base ester for the manufacture of a medicament for the treatment or prevention of the above-mentioned conditions and diseases. The invention also includes a method of treating or preventing such conditions and diseases comprising administering to a mammal, including a human, in need of such treatment an effective amount of a compound as defined above.

(4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl ester In a specific embodiment of the mesylate salt (i.e., methanesulfonate), the applicant has achieved a highly crystalline polymorph, the high crystallinity of which is herein derived from X-ray powder diffraction (XRPD). Confirmed with a polarizing microscope (PLM).

Figure 1 shows (4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid ( 3S )-tetrahydrofuran- XRPD of 3-based ester free base.

Figure 2 shows (4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid ( 3S )-tetrahydrofuran- XRPD of the crystal of 3-yl ester mesylate. The XRPD system is double-covered.

FIG 3 show (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran - 1H NMR spectrum of 3-yl ester free base (labeled as (A)) and hydrochloride (labeled as (B)).

FIG 4 show (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran - 1H NMR spectrum of 3-based ester free base (labeled as (C)) and phosphate (labeled as (D)).

The present inventors have studied (4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S ) in the long term. -tetrahydrofuran-3-yl ester and 22 kinds of acids (hydrochloric acid, sulfuric acid, 1,2-ethanedisulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, benzenesulfonic acid, L-aspartic acid, maleic acid, phosphoric acid , ethanesulfonic acid, L-glutamic acid, L-tartaric acid, fumaric acid, citric acid, L-malic acid, D-gluconic acid, D/L-lactic acid, L-lactic acid, benzoic acid, succinic acid, After the diacid and acetic acid were reacted to form a salt, four novel crystalline salt forms were found, namely: (4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4 , 5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran-3-yl ester hydrochloride; (4 S) -4- isopropyl-1,4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran-3-yl phosphate ester; (4 S) -4- isopropyl-1,4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran-3-yl ester sulfate hydrate; and (4 S) -4- isopropyl-1,4, 6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran-3-yl ester methanesulfonate.

It has been found that the free base and 1,2-ethanedisulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, L-aspartic acid, maleic acid, ethanesulfonic acid, L-glutamic acid, L-tartaric acid, rich Horse acid, citric acid, L-malic acid, D-gluconic acid, D/L- The salt formed by lactic acid, L-lactic acid, benzoic acid, succinic acid, adipic acid, and acetic acid is amorphous.

Four kinds of crystalline salts were tested to determine their ease of handling, hygroscopicity, and thermal stability. Hydrochlorides and phosphates have operational properties that are improved over free base glass/gel based on their crystallinity. However, preliminary experiments showed that the two salt forms still have some hygroscopicity. Both of these salts deliquesce after being stored overnight at 40 ° C in 75% relative humidity.

The mesylate and sulfate are based on their crystallinity and have improved handling properties over the free base gum. Both salts have surprisingly reduced hygroscopicity. The methanesulfonate was deliquescent after storage for 3 days in a 75% relative humidity environment at 40 °C. Sulfate remained unchanged after 7 days of storage at 75% relative humidity at 40 °C.

The advantage of mesylate and sulphate is the significantly improved thermal stability. The methanesulfonate has a melting point of 189 °C. The melting point of the sulfate is 106 °C. Based on these melting points, it is expected that both the mesylate salt and the sulfate salt can withstand the grinding and pressing process without melting or hindering the process. Thus, both the sulfate and the mesylate salt have surprisingly improved hygroscopicity and surprisingly improved thermal stability compared to the corresponding free base.

(4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c] by X-ray powder diffraction (XRPD) and polarized light microscopy (PLM) pyridine-5-carboxylic acid (3 S) - tetrahydrofuran-3-yl methanesulphonate ester of (i.e. methanesulfonate) is highly crystalline.

definition

This article uses "treatment" to prevent the disease of the name Or a condition, or alleviating or eliminating a condition once established.

"Effective amount" means the amount of a compound that imparts a medical benefit to an individual being treated. The medical effect can be objective (i.e., can be measured by certain tests or markers) or subjective (i.e., the effect or sensation presented by the individual).

"Pharmaceutically acceptable" means suitable for the preparation of a pharmaceutical composition which is generally safe, non-toxic and not biologically or otherwise unacceptable, and includes those suitable for veterinary use and human medical use.

Unless otherwise stated, the term "(4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c] is used herein to describe the salt form. pyridine-5-carboxylic acid (3 S) - tetrahydrofuran-3-yl ester "includes (3S, 4S) and (3R, 4R) enantiomers of the mixture. A specific example, (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - The tetrahydrofuran-3-yl ester and its salt have an absolute purity of >95%, preferably >99%, more preferably >99.5%. A specific example, (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - Tetrahydrofuran-3-yl ester means an (3S, 4S) enantiomer having an enantiomeric purity of >95%, preferably >99%, more preferably >99.5%. A specific example, (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - Tetrahydrofuran-3-yl ester has a diastereomeric purity of >95%, preferably >99%, more preferably >99.5%.

Composition

The compounds of the invention for clinical use are formulated into pharmaceutical formulations for a variety of different modes of administration. It is understood that the compounds of the present invention are Administration with a physiologically acceptable carrier, excipient, or diluent. The pharmaceutical composition of the present invention can be administered by any suitable route, preferably oral, rectal, nasal, topical (including buccal and sublingual), sublingual, transdermal, intrathecal, transmucosal or parenteral (including Subcutaneous, intramuscular, intravenous and intradermal).

Other formulations are preferably presented in unit dosage form, for example, lozenges and sustained release capsules, and are included in the vesicles and may be prepared by any of the methods known in the art. The pharmaceutical preparations are generally prepared by mixing active substances or pharmaceutically acceptable salts thereof with conventional pharmaceutically acceptable carriers, diluents or excipients. Examples of excipients are water, gelatin, gum arabic, lactose, microcrystalline cellulose, starch, sodium starch glycolate, calcium hydrogen phosphate, magnesium stearate, talc, cerium oxide colloid, and the like. These formulations may also contain other pharmaceutically active agents, as well as conventional additives such as stabilizers, wetting agents, emulsifiers, flavoring agents, buffers, and the like. In general, the active compound content is from 0.1 to 95% by weight of the preparation, preferably from 0.2 to 20% by weight in the parenteral preparation, and more preferably from 1 to 50% by weight in the oral administration preparation. .

The formulation can be further prepared by known methods such as granulation, compression, micro-embedding, spraying, and the like. The preparation can be prepared into a dosage form such as a tablet, a capsule, a granule, a powder, a syrup, a suspension, a suppository or an injection by a conventional method. Liquid formulations may be prepared by dissolving or suspending the active material in water or other suitable vehicle. Tablets and capsules can be applied in a conventional manner. In order to maintain a therapeutically effective plasma concentration for a long period of time, the compounds of the invention may be incorporated into a sustained release formulation.

The dose level and frequency of administration of a particular compound will vary with a variety of factors, including the potency of the particular compound employed, the metabolic stability and duration of action of the compound, the age, weight, general health, sex, diet, and mode of administration of the patient. It varies with time, rate of excretion, combination of drugs, severity of the condition being treated, and the therapy being performed by the patient. The daily dose may be, for example, in the range of from about 0.001 mg to about 100 mg per kg of body weight, and may be administered in a single dose or in multiple doses, for example, from about 0.01 mg to about 25 mg each. Usually, these doses are administered orally, but parenteral administration can also be selected.

experimental method

Salt formation experiment

The salt screening method was carried out using 24 acidic counterions (see Table 1), attempting to form both single and half salts, as appropriate. Nine sets of experiments were performed using different series of solvent systems and conditions.

Slow cooling to form a salt experiment

(4S)-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3S)-tetrahydrofuran-3-yl ester free The base (20 mg) sample was dissolved separately in IPA (2.5 vol), IPAc (2.5 vol), acetone-water (9:1 v/v, 2.5 vol) or DIPE (10 vol). The solution was heated to 40 ° C and each test acid (1 or 0.5 equivalents, see Table 1) was added with gentle stirring. The vial was kept at 40 ° C for 1 h and then cooled to 5 ° C at 1 ° C / min. The mixture was kept at 5 ° C overnight. All solids collected were collected by filtration and analyzed by XRPD. The oil quality and the gelatin are used for the ripening cycle: RT to 50 ° C, each temperature for 4 h, to facilitate crystallization. Allow the solution to evaporate under ambient conditions. The residue was analyzed by ¹ H-NMR and DSC to analyze the possible crystallization of salt formation and/or heating.

Add anti-solvent salt formation experiment

(4S)-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3S)-tetrahydrofuran-3-yl ester free The base (15 mg) sample was separately dissolved in IPAc (with and without 3% v/v water; 5 vol) and the solution was heated to 40 °C. Add the corresponding acid (1 or 0.5 equivalents, see Table 1) with gentle agitation. After the vial was kept at 40 ° C for 1 h, a gradually increasing amount of anti-solvent (n-heptane or TBME) was added until the solution turned cloudy. At this time, all samples were cooled to 5 ° C at 1 ° C / min, and kept at 5 ° C for the next night. If no precipitation occurs, add more anti-solvent. All solids collected were collected by filtration and analyzed by XRPD. Take the oily and colloid for ripening cycle, RT to 50 ° C, 4 h at each temperature to facilitate crystallization. If no precipitation occurs, allow the solution to cool below ambient temperature and allow evaporation.

Analytical method

X-ray powder diffraction (XRPD)

The X-ray powder diffraction pattern was applied to a Bruker AXS C2 GADDS diffractometer using Cu K α radiation (40 kV, 40 mA), an automated XYZ stage, a self-positioning laser laser microscope and a HiStar 2 dimensional area detector. The X-ray optics consists of a single Göbel multilayer mirror coupled to a 0.3 mm pinhole collimator. The performance of the instrument was tested weekly using a certified corundum standard (NIST 1976 Corundum) (plate). The beam divergence (i.e., the effective size of the X-ray beam on the sample) is about 4 mm. Using the θ-θ continuous scan mode, the distance between the sample and the detector is 20 cm, resulting in an effective 2 θ range of 3.2 ° - 29.7 °. Usually the sample will be exposed Under the X-ray beam for 120 seconds. The software used to collect the data was GADDS for WNT 4.1.16, and the data was analyzed using Diffrac Plus EVA v 9.0.0.2 or v 13.0.0.2. Samples that were run under ambient conditions were prepared as flat samples using powder received without grinding. Approximately 1-2 mg of the sample was lightly pressed onto the glass slide to obtain a flat surface. Samples that are operated under non-environmental conditions are placed on a wafer containing a thermally conductive compound. The sample was then heated at about 10 ° C/min to the appropriate temperature and then held at a constant temperature for about 1 min before data collection began.

Alternatively, the X-ray powder diffraction pattern is collected on a Bruker D8 diffractometer using Cu K alpha radiation (40 kV, 40 mA), θ-2 θ goniometer, V4 divergence and receiving slits, Ge monochromator, With the Lynxeye detector. The performance of the instrument was tested using a certified corundum standard (NIST 1976). The software used to collect the data was Diffrac Plus XRD Commander v2.5.0 and the data was analyzed and presented using Diffrac Plus EVA v 11.0.0.2 or v 13.0.0.2. The sample was used as a plate sample using the received powder and operated under ambient conditions. Approximately 20 mg of sample was gently loaded into the cavity cut into a polished zero background (510) wafer. The sample during the analysis is rotated in its own plane. The details of the collected data are - angle range: 2 to 42 ° 2 θ; step size: 0.05 ° 2 θ; collection time: 0.5 sec / step

Nuclear magnetic resonance (NMR)

^{The 1} H NMR spectrum was collected on an automated sampler equipped with a Bruker 400 MHz instrument controlled by a DRX400 console. Automated experiments were performed using ICONNMR v4.0.4 (version 1) operating with Topspin v 1.3 (patch level 10) using standard Bruker loading experiments. Non-routine spectral data is obtained by using only Topspin. Samples were prepared in d6-DMSO unless otherwise stated. Off-line analysis was performed using ACD SpecManager v 12.00 (version 29094). Alternatively, ¹ H NMR spectra were collected on a Bruker Avance III 400 MHz QNP Ultrashield Plus Cryo.

Liquid chromatography-mass spectrometry (LCMS)

Analytical grade LCMS was performed on an Agilent 1100 HPLC system coupled to a Waters ZQ mass spectrometer using a Phenomenex Synergi column (RP-Hydro, 150 x 4.6 mm, 4 um, 1.5 mL/min, 30 ° C, gradient 5-100% MeCN (+ An aqueous solution (+0.1% TFA) of 0.085% TFA) lasted 7 min - 0.5 min, 200-300 nm).

Differential Scanning Calorimetry (DSC)

DSC data was collected on a TA Instruments Q2000 equipped with a 50 position autosampler. Use sapphire to calibrate heat capacity and use certified indium to calibrate energy and temperature. Typically, 0.5-3 mg of each sample was heated from 25 ° C to 350 ° C at 10 ° C/min in a pinhole aluminum pan. The sample was purged with dry nitrogen at 50 mL/min. The DSC system for temperature regulation was carried out using a basic heating rate of 2 ° C/min and a temperature control parameter of ± 1.27 ° C / min and 60 seconds. The instrument control software was Advantage for Q Series v2.8.0.392 and Thermal Advantage v4.8.3, using Universal Analysis v4.3A to analyze the data.

Thermogravimetric analysis (TGA)

TGA data was collected on a TA Instruments Q500 TGA equipped with a 16 position autosampler. Use certified aluminum-nickel-manganese alloy (Alumel) with Nickel corrects the temperature of the instrument. Typically, 5-30 mg of each sample was loaded onto pre-stripped platinum rhodium and aluminum DSC pans and heated from ambient temperature to 350 °C at 10 °C/min. The sample was purged with nitrogen at 60 mL/min. The instrument control software is Advantage for Q Series v2.8.0.392 and Thermal Advantage v4.8.3.

Polarized light microscope (PLM)

The samples were studied with a Leica LM/DM polarizing microscope and images were acquired using a digital camera. Take a small amount of each sample on a glass slide, place it in the soaking oil, cover it with a cover slip, and separate each particle as much as possible. The samples were observed under coupled λ false color filter and appropriate magnification and partial polarization.

Hot plate microscope (HSM) [melting point]

The hot plate microscope was mounted on a Leica LM/DM polarizing microscope with a Mettler-Toledo MTFP82HT heating plate and a digital camera for image acquisition. Take a small amount of each sample on a glass slide and separate each particle as much as possible. The sample was observed under heating from ambient temperature with a λ false color filter coupled with appropriate magnification and partial polarization, typically at 10-20 ° C/min.

Chemical purity determination by HPLC

Purity analysis was performed on an Agilent HP1100 series system equipped with a diode array detector and using ChemStation software vB.02.01-SR1 using the method detailed below (Table 1).

Determination of palm purity by palmar HPLC

The palm HPLC was performed on an Agilent 1200 system using an Astec Chirobiotic T 100 x 4.6 mm 5um column, reversed phase polarity, 150 x 4.6 mm, 5 um, isocratic 85% MeOH 15% 20 mM ammonium acetate over 10 min, 1.0 mL /min, 220nm.

Determination of water by Karl Fischer Titration (KF)

The water content of each sample was on a Mettler Toledo DL39 Coulometer colorimeter using Hydranal Coulomat AG reagent with argon purge. Determination. A weighed solid sample is added to a container on a platinum TGA tray that is connected to a threaded subaseal (to avoid water infiltration). A 10 mg sample was used for each titration and a double replicate assay was performed.

Gravity Vapor Adsorption (GVS)

The adsorption isotherm curve was obtained using an SMS DVS inherent moisture adsorption analyzer controlled by DVS Intrinsic Control software v1.0.0.30. The sample temperature was maintained at 25 ° C by instrument control. The humidity was controlled using a dry and wet mixed nitrogen stream at a total flow rate of 200 mL/min. The relative humidity is measured by a calibrated Rotronic probe (1.0-100% RH dynamic range) located near the sample. The sample weight change (mass relaxation as a function of % RH) was continuously monitored by a microbalance (accuracy ± 0.005 mg). Typically 5-20 mg of the sample is placed in a de-stressed stainless steel basket under ambient conditions. The samples were loaded and unloaded at 40% RH and 25 ° C (typical room conditions). The moisture adsorption isotherm was completed as described below (1 full cycle for 2 scans). A standard isothermal curve was completed at 25 ° C at 10% RH intervals in the range of 0.5% to 90% RH. Data analysis was performed in Microsoft Excel using DVS Analysis Suite v6.0.0.7. Method parameters of SMS DVS intrinsic experiment: adsorption scan 1 40-90; desorption/adsorption scan 2 90-0, 0-40; interval (%RH) 10; scan number 4; flow rate (mL/min) 200 Temperature (°C) 25; stability (°C/min) 0.2; adsorption time (h) 6h cutoff. Samples were recovered after completion of the isothermal curve and reanalyzed using XRPD.

Ion chromatography (IC)

For use with Metrohm 761 Compact IC (for cations) and Metrohm 861 Advanced Compact IC (for anions), use IC Net software V2.3 collects data. Take the accurately weighed sample into a suitable solution to make a stock solution, first diluted 1:9 before use in the test. Quantification is performed as compared to a known concentration standard solution of the analyzed ions. IC method parameters for anion chromatography: method type - anion exchange; column - Metrosep A Supp 5-250 (4.0 × 250mm); column temperature (°C) ambient temperature; injection (μl) 20; detection - conductivity Detector; flow rate (mL/min) 0.7; dissolvate 3.2 mM sodium carbonate, 1.0 mM sodium bicarbonate in 5% acetone solution.

result

Isolation of crystalline hydrochloride, sulfate, phosphate and methanesulfonate, using some or all of XRPD, ¹ H NMR, DSC, TGA, GVS, IC, PLM, HSM, HPLC and KF analysis of their characteristics ( See Table 2). Hydrochloride and phosphate are highly hygroscopic. Further analysis of the mesylate salt and sulfate.

(4S)-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3S)-tetrahydrofuran-3-yl ester, and Synthesis of its mesylate salt and sulfate form:

The following abbrils have been used:

experimental method

All reagents were on a commercial grade and were used without further purification unless otherwise stated. Reagent grade solvents were used for all examples. Analytical grade LCMS was performed on a Waters ZQ mass spectrometer connected to an Agilent 1100 HPLC system. Analytical HPLC was performed on an Agilent 1100 system. High resolution mass spectrometry (HRMS) was obtained from an Agilent MSD-TOF coupled to an Agilent 1100 HPLC system. During the analysis, if necessary, two quality check corrections are used and automatically corrected. The map is obtained according to the positive electric spray mode. The mass range obtained is m/z 100-1100. A pattern detection method using a quality peak. Flash chromatography is performed on a CombiFlash Companion system equipped with a RediSep gangue column or a Flash Master Personal system equipped with Strata SI-1 meteorite Gigatubes. Reverse HPLC was performed on a Gilson system equipped with a Phenomenex Synergi Hydro RP 150×10 mm, YMC ODS-A 100/150×20 mm or Chirobiotic T 250×10 mm column (Gilson 322 equipped with Gilson 321 balanced pump and Gilson 215 autosampler) On the pump). Reverse phase column chromatography was performed on a Gilson system (Gilson 321 Pump and Gilson FC204 Dissolution Collector) equipped with a Merck LiChroprep ^® RP-18 (40-63 μm) vermiculite column. The compounds were automatically named using ACD 6.0. All compounds were dried overnight in a vacuum oven.

Analytical HPLC and LCMS data were obtained from: System A: Phenomenex Synergi Hydro RP (C18, 30 x 4.6 mm, 4 μm), gradient 5-100% CH ₃ CN (+0.085% TFA) in water (+0.1% TFA) , 1.5 mL/min, gradient time 1.75 min, 200 nm, 30 ° C; or System B: Phenomenex Synergi Hydro RP (C18, 150 x 4.6 mm, 4 μm), gradient 5-100% CH ₃ CN (+0.085% TFA) Aqueous solution (+0.1% TFA), 1.5 mL/min, gradient time 7 min, 200 nm, 30 °C.

The palmar HPLC data was obtained from: System C: Chirobiotic V polar ionic mode (150 x 4.6 mm), 70% MeOH in 10 mM ammonium formate buffered water, 1.0 mL/min over 10 min, 200 nm, 30 °C.

Intermediate 1

4-isopropyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine hydrochloride

The histamine dihydrochloride (61.9 g, 336 mmol) was dissolved in NaOH (33.6 g, 841 mmol) in water (125 mL) and MeOH (500 mL). Aldehyde (61.4 mL, 672 mmol). The reaction mixture was heated at reflux <RTI ID=0.0></RTI> to <RTI ID=0.0></RTI> </RTI> <RTIgt; The residue was dissolved in warm MeOH (300 mL)EtOAc. The residue was stirred with MeOH (50 mL) &EtOAc. The resulting precipitate was filtered and washed with acetone (100 mL) to yield 4-isopropyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine hydrochloride (33.0 g, 48.7 %) white solid.

Analytical grade LCMS: purity >90% (System A, R _T = 0.51 min), ES ⁺ : 166.4 [MH] ⁺ .

Intermediate 2

4-isopropylphenyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid 4-nitrophenyl ester

Intermediate 1 (2.78 g, 8.28 mmol, 60% purity) and DIPEA (5.27 mL, 30.3 mmol) were taken in DCM (100 mL). The reaction mixture was cooled to 0 ° C, and 4-nitrophenyl chloroformate (4.07 g, 20.2 mmol). The reaction mixture was stirred at room temperature for 18 h. The reaction mixture with saturated aqueous NaHCO ₃ (5 × 100mL) washed, dehydrated (MgS04 _4), and the solvents were removed in vacuo to produce 4-isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4, Yellow gum of 5-c]pyridine-5-carboxylic acid 4-nitrophenyl ester (5.28 g, crude product).

Analytical grade HPLC: purity 41% (system B, R _T = 4.70 min); analytical grade LCMS: purity 86% (system A, R _T = 1.70 min), ES ⁺ : 331.0 [MH] ⁺ .

(4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl ester

NaH (0.40 g, 10.0 mmol, 60% homogenate in mineral oil) was suspended in anhydrous THF (20 mL), cooled to 0 ° C, and ( S )-3-hydroxytetrahydrofuran (0.88 g, 0.68 mL, 10.0 mmol). The suspension was stirred at 0&lt;0&gt;C for 30 min then EtOAc (EtOAc)EtOAc. After 5 and 29 h, respectively, two additional portions of NaH and THF containing ( S )-3-hydroxytetrahydrofuran were added. After 2 days, water (10 mL) was added to the reaction mixture to quench the reaction and solvent was evaporated in vacuo. The residue was dissolved in EtOAc (100mL), using 1M Na ₂ CO ₃ solution (4 × 100mL) washed, dehydrated (MgSO _4), and the solvents were removed in vacuo. The residue was purified by column chromatography (normal phase, 20 g, Strata SI-1, vermiculite Gigatube, DCM (200 mL), followed by a solution of 2%, 4% and 5% MeOH in DCM (200 mL) and reverse phase HPLC (YMC ODS-A 100×20 mm, 5 μm, 25 mL/min, gradient 30% to 60% over 7 min, then 100% (3 min) MeOH in 10% MeOH/aq) to yield 4-isopropyl-1 , 4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran-3-yl ester (34.8mg, 1.1%) of a white solid.

Analytical grade HPLC: purity 100% (system B, R _T = 3.63 min); analytical grade LCMS: purity 100% (system B, R _T = 4.01 min), ES ⁺ : 280.1 [MH] ⁺ .

4-isopropyl-l, 4,6,7-tetrahydro take -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran-3-yl ester (39.91mg) Dissolved in 10 mM ammonium formate buffer and MeOH (2 mL, 1:1), using reverse relative palm chromatography HPLC (Chirobiotic T 250×10 mm, 3 mL/min, isocratic operation of 70% MeOH in 10 mM ammonium formate buffer solution (40 min) ), pH 7.4) was purified 2 times, to produce a single diastereomer thereof, (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c ] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran-3-yl ester (6.90mg, 99% ee).

Analytical HPLC: purity 100% (system B, R _T = 3.63 min); palmar HPLC: purity 99.5% (system C, R _T = 2.22 min); analytical grade LCMS: purity 100% (system B, R _T ^{= 3.90min), ES +: 280.1} [MH] +; HRMS C 14 H 21 N 3 O 3: calcd 279.1583, found 279.1571.

(4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl ester Methanesulfonate

(4S)-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3S)-tetrahydrofuran-at room temperature The 3-yl ester free base (460 mg, 1.65 mmol) was dissolved in EtOAc (10 mL). Methanesulfonic acid (107 uL) was added in portions under gentle heating. The solution was allowed to cool to room temperature overnight. The resulting crystals were collected by chromatography, washed with EtOAc (EtOAc) (4S)-4-Isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3S)-tetrahydrofuran-3-yl ester A Sulfonate, 99% yield (615 mg) of a white solid. HPLC: retention time 2.27 min, purity 99.5%. Melting point: 189 ° C. LCMS: residence time 4.19 min, ES ⁺ 280.0 [MH] ⁺ , 100% purity. For palm HPLC: retention time 3.70 min, >99.5% de. ¹ H NMR (400 MHz, CDCl ₃ ): δ _H 8.72 (1H, m, NHH H NH ⁺ ), 5.29 (1H, m, OC H ), 5.05 (0.5H, d, J 8.4 Hz, CC H N), 4.89 (0.5H, d, J7.6Hz, CC H N), 4.59 (0.5H, m, NC H _A CH _B ), 4.39 (0.5H, m, NC H _A CH _B ), 3.97-3.85 (4H, m, C H ₂ OC H ₂ ), 3.20 (1H, m, NCHAC H _B ), 2.89 (3H, s, C H ₃ SO ₃ ^- ), 2.89-2.72 (2H, m, CC H ₂ CH ₂ N) , 2.23-2.07 (3H, m, C H (CH 3) 2, OCH 2 C H 2), 1.16 (3H, d, J 6.4Hz, C H 3) and 1.06-0.96 (3H, m, C H 3 ).

(4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl ester Sulfate

(4S)-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3S)-tetrahydrofuran-3-yl ester free Base (440.4 mg, 1.6 mmol) was dissolved in EtOAc (5 vol; 2.20 mL). The clear solution was heated to 40 ° C and maintained at this temperature for 30 min. H ₂ SO ₄ (1M in THF, 1 eq., 1.6 mL) was then added with stirring to give a white solid. This suspension was set to cool to 5 ° C at 1 ° C/min and maintained at this temperature for 20 h. The solid was filtered off with suction and dried under vacuum at room temperature overnight. (4S)-4-Isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3S)-tetrahydrofuran-3-yl ester sulfate Salt, 65% yield, white solid. The HPLC purity was 99.3%. Melting point: 106 ° C. _{^{1 H NMR (400MHz, CDCl 3}} ): δ H 13.70 (1H, br, H SO 4 -), 9.05 (1H, m, NHC H NH +), 5.28 (1H, m, OC H), 4.95 (0.5H ,d,J 8.4Hz,CC H N),4.86(0.5H,d,J 7.6Hz,CC H N),4.55(0.5H,m,NC H _A CH _B ),4.35(0.5H,m,NC H _A CH _B ), 3.95-3.74 (4H, m, C H ₂ OC H ₂ ), 3.15 (1H, m, NCH _A C H _B ), 2.78-2.67 (2H, m, CC H ₂ CH ₂ N) , 2.21-1.99 (3H, m, C H (CH ₃ ) ₂ , OCH ₂ C H ₂ ), 1.09 (3H, d, C H ₃ ) and 0.94-0.81 (3H, m, C H ₃ ).

Stability/hygroscopicity during long-term storage

Also Analysis (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran-3 The stability and hygroscopicity of the ester methanesulfonate was taken from a 3 g portion added to a double LDPE mat, sealed with a drawstring and a desiccant package was added to the foil pouch, followed by heat sealing. The foil pouch is then placed in an HDPE bucket with an HDPE lid. These conditions simulate typical GMP grade storage conditions. The stability was analyzed by HPLC and the hygroscopicity was analyzed by Karl Fischer (KF) titration. (4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl ester The mesylate salt degraded only 0.1% at 25 ° C / 60% RH for 3 years, did not absorb water, only degraded 0.1% after 6 months at 40 ° C / 75% RH, only water absorption of 0.1%.

Single crystal X-ray diffraction (SXRPD)

The data was collected on an Oxford Diffraction SuperNova dual source diffracted, zero copper, Atlas CCD diffractometer equipped with an Oxford Cryosystems Cobra cooling unit. The data was collected using CuK alpha radiation. The structure is typically parsed using a SHELXS or SHEMLD program and refined using the SHELXTL program that is part of the Bruker AXS SHELXTL suite of software. Unless otherwise stated, the carbon-bonded hydrogen atoms are geometrically placed and can be refined using a riding isotropic displacement parameter. The hydrogen atoms attached to the heteroatoms are positioned by differential Fourier synthesis and can be freely refined with an isotropic displacement factor.

Single crystal structure determination

(4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl ester The single crystal of the mesylate salt was slowly evaporated from ethyl acetate/methanol to grow.

The structural analysis is obtained by a direct method, and the full-matrix least-squares refinement is performed by F ² , and the weight w ⁻¹ =σ ² ( F _o ² )+(0.0490 P ) ² +(0.3000 P ), where P = ( F _o ² +2 Fc ² )/3, anisotropic shift parameter. Using the spherical harmonic function, the SCALE3 ABSPACK scaling algorithm is substituted for empirical absorption correction.

Absolute structural parameter = 0.002 (13). All data is ultimately ^{wR 2 = {Σ [w (} F o 2 - Fc 2) 2] / Σ [w (F o 2) 2] 1/2} = 0.0657, F o> 4 σ (F o) of 2651 the F value of the reflected common R ₁ = 0.024, 234 all data associated with the parameter S = 1.007. Finally Δ / σ (maximum) 0.000, △ / σ (average value) 0.000. The final difference spectrum is between +0.24 and -0.288e Å ^-3 .

The analytical crystal structure parameters are as follows (Table 3):

X-ray powder diffraction

(4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl ester The X-ray diffraction pattern of the methanesulfonate powder is 8.549, 9.851, 10.899, 12.295, 13.198, 13.393, 14.083, 15.897, 16.280, 17.097, 17.744, 18.289, 19.694, 20.180, 20.443, 20.597, 20.886, 21.948, 22.112. Waves appear at 22.444, 23.194, 23.653, 24.144, 24.714, 25.292, 25.590, 25.810, 26.526, 26.765, 27.084, 27.283, 27.662, 28.159, 28.996, 29.135, 29.912 and 30.868 degrees 2 θ. Considering normal experimental variations, the peaks identified above should be considered to be +/- 0.2 degrees 2 θ, such as: +/- 0.1, +/- 0.05, +/- 0.01, +/- 0.005, and +/ - 0.001. The relative intensities of these peaks are as follows ( Table 4 ):

Other feature analysis

(4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl ester The 1H NMR spectrum of the free base (upper line, labeled (A)) and the corresponding hydrochloride (lower line, labeled (B)) is shown in Figure 3.

(4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl ester The 1H NMR spectrum of the free base (upper line, labeled (C)) and the corresponding phosphate (lower line, labeled (D)) is shown in Figure 4.

Since the figure in this case is only test data, it is not a representative figure of this case.

Therefore, there is no designated representative map in this case.

Claims (25)

(4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl Ester mesylate, hydrated therewith. (4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl Ester sulfate, hydrated therewith. (4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylic acid (3 S )-tetrahydrofuran-3-yl Ester sulfate 1.5 H ₂ O. A crystalline (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] - pyridine-5-carboxylic acid (3 S) - tetrahydrofuran -3 a hydroxy ester mesylate having a space group P2(1)2(1)2(1) , and a unit cell size substantially as follows: A crystalline (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] - pyridine-5-carboxylic acid (3 S) - tetrahydrofuran -3 a mesylate salt of a base ester having an XRPD pattern comprising the following 2 theta value measured using CuK alpha radiation: 21.948. A salt as described in claim 5, having an XRPD pattern comprising the following 2 theta values measured using CuK alpha radiation: 17.744 and 21.948. A salt as described in claim 5 or 6, which has an XRPD pattern comprising the following 2 θ values measured using CuK α radiation: 17.744, 20.886, 21.948. A salt according to any one of claims 5 to 7 which has an XRPD pattern comprising the following 2 θ values measured using CuK α radiation: 17.744, 20.886, 21.948, and 9.851. A salt according to any one of claims 5 to 8 which has an XRPD pattern comprising the following 2θ values measured using CuK α radiation: 17.744, 20.886, 21.948, 9.851 and 16.280. A salt according to any one of claims 5 to 9 which has an XRPD pattern comprising the following 2θ values measured using CuK α radiation: 9.851, 16.280, 17.097, 17.744, 19.694, 20.443, 20.886, 21.948, 22.112, 23.194, 23.653, 24.144, 27.084, 27.283 and 29.912. A salt according to any one of claims 5 to 10, which has an XRPD pattern as shown in Table 4 and/or Figure 2. The salt of any one of claims 1 to 11 which has a purity greater than 95%. A salt according to any one of claims 1 to 11, which has a purity higher than 99%. The salt of any one of claims 1 to 11, which has a purity higher than 99.5%. The salt of any one of claims 1 to 14 which has an enantiomeric purity of greater than 95%. The salt of any one of claims 1 to 14 which has an enantiomeric purity of greater than 99%. The salt according to any one of claims 1 to 14, which has It has an enantiomeric purity higher than 99.5%. A pharmaceutical composition comprising the salt of any one of claims 1 to 17 in combination with one or more suitable excipients. A salt according to any one of claims 1 to 17, or a pharmaceutical composition according to claim 18, which is for use in the treatment of a disease selected from the group consisting of an inflammatory, an inflammatory disease, an immune or an autoimmune disease. A disease or condition, or inhibiting tumor growth, or for the manufacture of a medicament for treating a disease or condition selected from the group consisting of an inflammatory, inflammatory, immune or autoimmune disorder, or inhibiting tumor growth. A method of treating a disease selected from an inflammatory, inflammatory, immune or autoimmune disease, or inhibiting tumor growth, comprising administering an effective amount to an individual suffering from such a disease as claimed in claims 1 to 17 A salt as described, or a pharmaceutical composition as described in claim 18 of the patent application. A salt according to claim 17, or a pharmaceutical composition according to claim 18, or the method of claim 20, wherein the inflammatory or inflammatory disease or immunity or self Physical immune disorders are arthritis (including rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis and dry arthritis), synovitis, vasculitis, Sjogren's disease, and inflammation of the intestines Disorders (including Crohn's disease, ulcerative colitis, inflammatory bowel disease and intestinal bowel disease), atherosclerosis, multiple sclerosis, Alzheimer's disease, vascular dementia, Parkinson's disease Parkinson's disease, cerebral amyloid angiopathy, somatic cerebral arterial disease Complicated with subcortical cerebral infarction and white matter lesions, pulmonary inflammatory diseases (including asthma, chronic obstructive pulmonary disease and acute respiratory distress syndrome), fibrotic diseases (including idiopathic pulmonary fibrosis, cardiac fibrosis, liver fibrosis) And systemic sclerosis (scleroderma), skin inflammatory diseases (including contact dermatitis, atopic dermatitis and dryness), eye inflammatory diseases (including age-related macular degeneration, uveitis and diabetic retinopathy) Systemic inflammatory response syndrome, sepsis, inflammation of the liver and/or autoimmune disorders (including autoimmune hepatitis, primary biliary cirrhosis, alcoholic liver disease, sclerosing cholangitis, and autoimmune bile ducts) Inflammation, diabetes (type I or II) and/or its complications, chronic heart failure, congestive heart failure, ischemic disease (including stroke and ischemia-reperfusion injury) or myocardial infarction and/or its complications Or epilepsy. The salt according to claim 17, or the pharmaceutical composition according to claim 18, or the method according to claim 20, which is used for treatment selected from the group consisting of rheumatoid Arthritis, osteoarthritis, liver fibrosis, chronic obstructive pulmonary disease, multiple sclerosis, Sjogren's disease, Alzheimer's disease, Parkinson's disease, inflammatory bowel disease, Or a disease of vascular dementia. A method of manufacturing crystalline (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran -3 - method of the methanesulfonate ester, a system which is (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] pyridine-5 carboxylic acid (3 S) - tetrahydrofuran-3-yl ester free base form of the methanesulfonate. The method of claim 23, wherein the methanesulfonate is added with methanesulfonic acid to (4 S )-4-isopropyl-1,4,6,7-tetrahydro-5H-imidazole. [4,5-c] pyridine-5-carboxylic acid (3 S) - tetrahydrofuran-3-yl ester free base as the formers. A crystalline (4 S) -4- isopropyl-l, 4,6,7-tetrahydro -5H- imidazo [4,5-c] - pyridine-5-carboxylic acid (3 S) - tetrahydrofuran -3 A hydroxyester mesylate salt obtained by the process described in claim 23 or 24.