Crystalline forms of plasma kallikrein inhibitors

Abstract

To provide a compound useful for treating plasma kallikrein-dependent diseases or conditions.SOLUTION: The present invention provides pure crystalline forms of 1-benzyl-N-(4-carbamimidoylbenzyl)-1H-pyrazole-4-carboxamide acetate, and an amorphous form, pharmaceutical compositions thereof, and methods for making the same.SELECTED DRAWING: None

Description

Technical Field

[0001] [Cross - Reference to Related Applications] This application claims priority to U.S. Provisional Application No. 62 / 871,517, filed Jul. 8, 2019, which is hereby incorporated by reference in its entirety for all purposes.

[0002] The present invention relates to crystalline forms of plasma kallikrein inhibitors, methods for producing polymorphs of plasma kallikrein inhibitors substantially free of other polymorphs, and compositions comprising substantially pure polymorphs of plasma kallikrein inhibitors.

Background Art

[0003] Thrombosis is essential to prevent blood loss and enable the repair of damaged blood vessels, a process known as hemostasis. However, if a thrombus occludes a blood vessel and deprives tissues of oxygen, it can become pathological. Arterial thrombosis, the occlusion of an artery by a thrombus, most commonly occurs at the site of atherosclerotic plaques that have ruptured or ulcerated (V. Kou et al., Mt Sinai J Med (2006) 73:449 - 68). Specific occlusion of the coronary arteries results in acute coronary syndromes, including unstable angina and myocardial infarction (MI).

[0004] Fibrin clots can be generated by the activation of either of two different pathways, the intrinsic and extrinsic pathways, which converge on a common coagulation pathway (R.G. Macfarlane, Nature (1964) 202:498 - 99; E.W. Davie et al., Science (1964) 145:1310 - 12; K. Joseph et al., Adv Immunol (2005) 86:159 - 208). Experimental data suggest that both plasma kallikrein (PK)-deficient and factor XII (FXII)-deficient individuals, although having no bleeding phenotype, have a major defect in clot formation via the intrinsic pathway (O.D. Ratnoff et al., J Clin Invest (1955) 34:602 - 13; R.W. Colman, (2001) in “Hemostasis and Thrombosis: Basic Principles and Clinical Practice” (R.W. Colman et al., eds., Lippincott, Williams & Wilkins, Philadelphia, Pa., pp.103 - 122); E.D. Rosen et al., Nature (1997) 390:290 - 94; W.E. Hathaway et al., Blood (1965) 26:521 - 32; A.S. Lawrie et al., Clin Lab Haematol (1998) 20:179 - 86; and S.M. Bates et al., Circulation (2005) 112:53 - 60). In the intrinsic pathway, binding to a surface activates a small amount of FXII (FXIIa), which then activates PK through proteolysis. Importantly, PK subsequently generates more FXIIa within a feedback loop, which then activates factor XI (FXI) to FXIa, linking to the common pathway.The initial activation of the intrinsic pathway is through a small amount of FXIIa that activates a small amount of PK. However, it is the subsequent feedback activation of FXII by PK that controls the degree of activation of the intrinsic pathway, and thus it is downstream coagulation (W.E. Hathaway et al., Blood (1965) 26:521-32).

[0005] Current treatments for acute MI or ischemic stroke in the hospital setting require emergency measures to dissolve occlusive thrombi and enable reperfusion (restoration of blood flow). One common way to do this is by administering to the patient a fibrinolytic drug such as tissue plasminogen activator (t-PA) or streptokinase, which generates active plasmin from plasminogen. Plasmin cleaves the fibrin mesh of the thrombus and performs clot lysis. Such fibrinolytic drugs are the most frequently used therapeutic agents for reperfusion worldwide. However, thromboembolic recurrence is also highly associated with fibrinolytic therapy, and subsequent reocclusion rates can be as high as 50% in some studies (F. Zijlstra et al., N Engl J Med (1993) 328:680-84; B.R. Brodie et al., Circulation (1994) 90:156-62; G.W. Stone et al., Circulation (1999) 99:1548-54; H. Tamai et al., Am Heart J (2004) 147:E9; F.W. Verheugt et al., J Am Coll Cardiol (1996) 27:766-73).

[0006] Patients who have experienced an acute MI show clinical evidence of a hypercoagulable (procoagulant) state. This hypercoagulability paradoxically worsens in those undergoing fibrinolytic therapy. In such treated patients, an increase in thrombin generation is observed, as assessed from thrombin-antithrombin III (TAT) levels that are up to two-fold higher compared to the already high levels observed in those receiving only heparin (H.M. Hoffmeister et al., Circulation (1998) 98:2527-33). The increase in thrombin is proposed to be due to activation of the intrinsic pathway via plasmin. Activation of the intrinsic pathway system via plasmin is known to occur in blood (G.A. Ewald et al., Circulation (1995) 91:28-36), and it has been suggested that this occurs as a result of direct activation of FXII by plasmin.

[0007] It is not only fibrinolytic-induced hypercoagulability that leads to an increased rate of reocclusion; it is also likely that at least in part, it is due to the major weakness of fibrinolytic therapy in not being able to achieve complete fibrinolysis of the thrombus (E.C. Keeley et al., Lancet (2003) 361:13-20). Another problem with fibrinolytic therapy is that it is associated with a three-fold increased risk of intracranial hemorrhage (ICH) (V. Menon et al., Chest (2004) 126:549S-575S; Fibrinolytic Therapy Trialists’ Collaborative Group, Lancet (1994) 343:311-22). Therefore, an adjunctive anticoagulant therapy that does not increase the risk of bleeding but inhibits the formation of new thrombin would be highly beneficial.

[0008] Administration of an irreversible inhibitor of FXII to wild-type mice has been found to lead to a decrease in occluded blood vessels and a reduction in ischemic cortical damage, and inhibition of FXII is protective against arterial thrombosis such as that occurring during acute MI or thrombotic stroke (WO 2006 / 066878A1 pamphlet). However, peptide pharmaceuticals have numerous weaknesses, including limitations in their application to acute tests due to their short half-life, intravenous administration requiring medical intervention, and the generation of anti-peptide antibodies by patients receiving treatment.

[0009] Plasma kallikrein has also been implicated in diabetic macular edema and diabetic retinopathy (A. Clermont et al., Diabetes (2011) 60:1590-98; J. A. Phipps et al., Hypertension (2009) 53:175-81); hereditary angioedema due to C1 inhibitor deficiency (A. Banerji et al., N Engl J Med (2017) 376:717-28; E. Aygoren-Pursun et al., N Engl J Med (2018) 379(4):352-62); acute liver injury (M. Li et al., Biochem Biophys Res Commun (2018) 504(4):857-64); inflammation and anaphylaxis (L. Bender et al., Front Immunol (2017) 8:1115); hemorrhagic changes and exacerbation of cerebral edema after treatment with recombinant tPA (F. Simao et al., Blood (2017) 129(16):2280-90); and chemically induced kidney injury (H. Wang et al., J Immunotoxicol (2016) 13(4):567-79).

[0010] The behavior of drug polymorphs can have significant implications in pharmacology. Polymorphs are different solid forms of the same molecule and can have different physical properties as a result of the arrangement of molecules in the crystal lattice. These different properties can affect pharmaceutical parameters such as storage stability, compressibility, density, hygroscopicity, dissolution rate, and bioavailability. It is also often possible to convert from one polymorph to another, and in some cases this occurs spontaneously. Storage stability can be affected if one polymorph can convert to a different polymorph with different density or hygroscopicity (which can cause tablets to expand or break). One polymorphic form of a compound may dissolve more readily or dissolve rapidly and well compared to another, increasing bioavailability. If the polymorph converts to a form with low bioavailability, the efficacy of the dosage form may decrease, and if it converts to a form with high bioavailability, the toxicity limit of the drug may be exceeded. Furthermore, the physical properties of the form can affect manufacturing, for example, one polymorph may be more likely to form a solvate or may be difficult to filter and wash free of impurities.

[0011] Suitable plasma kallikrein inhibitors have been developed (see Sinha et al., WO 2008 / 016883 pamphlet; US Patent No. 8,258,170). However, the identification of the crystalline forms of such compounds and the suitability of the formulations in the solid form have not yet been reported. Since the solubility, density, stability, and bioavailability of different crystalline forms may be different, there remain unaddressed requirements for the identification of such forms. SUMMARY OF THE INVENTION

[0012] Provided herein are pure crystalline forms, amorphous forms, and methods for producing each form of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate.

[0013] One aspect of the present invention is a crystalline form of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate (Compound 1), which may be in Form I, Form II, Form III, or Form IV, and this crystalline form substantially does not contain any other polymorphic forms.

[0014] Another aspect of the present invention is an amorphous form of Compound 1 (Form V), and this amorphous form substantially does not contain any crystalline forms.

[0015] Another aspect of the present invention is a pharmaceutical composition comprising a therapeutically effective amount of a pure solid form of Compound 1 (Form I, Form II, Form III, Form IV, or Form V) together with a pharmaceutically acceptable carrier.

[0016] Another aspect of the present invention is a method for producing Form I that substantially does not contain any other crystalline forms.

[0017] Another aspect of the present invention is a method for producing Form II that substantially does not contain any other crystalline forms.

[0018] Another aspect of the present invention is a method for producing Form III that substantially does not contain any other crystalline forms.

[0019] Another aspect of the present invention is a method for producing Form IV that substantially does not contain any other crystalline forms.

[0020] Another aspect of the present invention is a method for producing Form V that substantially does not contain any other crystalline forms.

[0021] Another aspect of the present invention is a method for treating a disease mediated by plasma kallikrein by administering the pharmaceutical composition of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0022]

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DETAILED DESCRIPTION OF THE INVENTION

[0023] Definitions Unless otherwise specified, the following terms used in this specification and the claims have the meanings set forth below.

[0024] The term "alkyl", unless otherwise specified, means a straight-chain or branched-chain hydrocarbon group having the specified number of carbon atoms (i.e., C1-C8 means 1 to 8 carbons), either alone or as part of another substituent. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. For each definition in this specification (e.g., alkyl, alkoxy, alkylamino, alkylthio, alkylene, haloalkyl), when a prefix indicating the number of main-chain carbon atoms in the alkyl portion is not included, the group or portion thereof has 12 or fewer main-chain carbon atoms.

[0025] As used herein, the term "composition" is intended to encompass a product containing the specified components in the specified amounts, and any product directly or indirectly resulting from a combination of the specified amounts of the specified components.

[0026] The term "polymorphic form" refers to the crystalline or amorphous solid form of a compound. Multiple polymorphic forms have different orientations or arrangements of molecules within the crystal structure. Different arrangements can result in differences in intermolecular forces, which can generate differences in melting point, solubility, hardness, stability, bioavailability, chemical reactivity, and the like. Polymorphic forms of the same compound can be identified by these properties and by their X-ray diffraction patterns. Four crystalline polymorphic forms and an amorphous form of compound 1 have already been prepared.

[0027] The term "amorphous" refers to a solid form having at most a limited degree of crystallinity as determined by XRPD. In an amorphous solid, the atoms are present in a disordered structure. In a crystalline substance or in a crystalline region, the atoms have both long-range and short-range order. In contrast, an amorphous substance has only short-range order. The degree of crystallization of the active substance can be measured using X-ray diffraction methods, including, for example, dynamic differential thermal measurements or XRPD.

[0028] The term "solid form" means any crystalline or amorphous form of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate, and mixtures thereof.

[0029] The terms "compound of formula I" and "compound 1" refer to 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate salt.

[0030] As used herein, the term "Form I" refers to the pure anhydrous crystalline form of compound 1. The XRPD diffraction pattern includes the following characteristic peaks: 10.0, 18.1, 18.6, 20.1, and 23.9 degrees ± 0.5, ± 0.2, or ± 0.1 degrees, 2θ, and this XRPD is made using CuKα radiation. The complete diffraction pattern for Form I is shown in Figure 1. A "pure" crystalline form substantially does not contain any other crystalline form.

[0031] As used herein, the term "Form II" refers to the pure hydrated crystalline form of compound 1. The XRPD diffraction pattern includes the following characteristic peaks: 4.2, 5.6, 8.4, 17.8, and 19.8 degrees, ± 0.5, ± 0.2, or ± 0.1 degrees, 2θ, and this XRPD is made using CuKα radiation. The complete diffraction pattern for Form II is shown in Figure 2.

[0032] As used herein, the term "Form III" refers to another pure crystalline form of Compound 1. The XRPD diffraction pattern includes the following characteristic peaks: 7.7, 10.7, 20.1, 23.7, and 24.3 degrees, ±0.5, ±0.2, or ±0.1 degree, 2θ, and this XRPD is made using CuKα radiation. The complete diffraction pattern for Form III is shown in Figure 3.

[0033] As used herein, the term "Form IV" refers to another pure crystalline form of Compound 1. The XRPD diffraction pattern includes the following characteristic peaks: 18.5, 19.6, and 23.7 degrees ±0.5, ±0.2, or ±0.1 degree, 2θ, and this XRPD is made using CuKα radiation. The complete diffraction pattern for Form IV is shown in Figure 4.

[0034] As used herein, the term "Form V" refers to a pure amorphous form of Compound 1. The XRPD diffraction pattern contains few or no characteristic peaks. The complete diffraction pattern for Form V is shown in Figure 5.

[0035] As used herein, the term "substantially free of" refers to a solid form of Compound 1 having at most a limited amount of different polymorphs. In some embodiments, a solid form of Compound 1 contains less than 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, or 0.01 weight % of other polymorphic forms, and in such a case, the solid form of Compound 1 is substantially free of other polymorphic forms. For example, if Form I consists of less than about 10 weight %, less than about 5 weight %, or less than about 1 weight % of Form II, Form III, Form IV, and / or Form V, then Form I is substantially free of other polymorphic forms. The purity of such forms can be measured using XRPD, differential scanning calorimetry, thermogravimetric analysis, and other methods known in the art.

[0036] As used herein, the term "subject" includes animals such as mammals including, without limitation, primates (including humans), cattle, pigs, sheep, goats, horses, dogs, cats, rabbits, rats, mice, and the like.

[0037] The term "effective amount" refers to an amount that provides a therapeutic benefit in the treatment, prevention, or management of a plasma kallikrein-mediated pathological process or an obvious symptom of a plasma kallikrein-mediated pathological process. The specific amount that is effective can be readily determined by a conventional physician and may vary depending upon, for example, the type of pathological process, the patient's medical history and age, the stage of the pathological process, and factors known in the art such as the like. Inhibition of plasma kallikrein can be measured, for example, without limitation, using an in vitro colorimetric assay or a fluorogenic substrate cleavage assay, or by measuring the clotting time of ex vivo whole blood or plasma (see, e.g., U.S. Patent No. 8,258,170).

[0038] General Compound 1 is useful as a plasma kallikrein (PK) inhibitor for the prevention and treatment of plasma kallikrein-dependent diseases or conditions, including blood coagulation disorders such as thrombosis, as well as other PK-dependent diseases and conditions. For example, this compound inhibits the formation of thrombin via the intrinsic pathway, thus reducing the risk of new pathogenic thrombus formation (reocclusion), and also improves reperfusion induced by fibrinolysis when administered as adjuvant therapy with a fibrinolytic regimen. Compound 1 is also useful for treating other diseases and disorders mediated by plasma kallikrein, such as, for example, but not limited to, diabetic macular edema, diabetic retinopathy, hereditary angioedema due to C1 inhibitor deficiency, acute liver injury, inflammation and anaphylaxis, hemorrhagic changes and exacerbation of cerebral edema after treatment with recombinant tissue plasminogen activator (tPA), chemically-induced kidney injury, ischemic stroke, hemorrhagic stroke, hypertension and its vascular complications (including retinopathy and nephropathy), cerebral vasogenic edema, pulmonary hypertension, inflammation, pain, acute myocardial infarction (MI), deep vein thrombosis (DVT), complications resulting from fibrinolytic therapy following stroke or MI (e.g., due to tissue plasminogen activator, streptokinase), angina, angioedema, sepsis, arthritis, complications of cardiopulmonary bypass, capillary leak syndrome, inflammatory bowel disease, diabetes and its vascular complications (retinopathy, diabetic macular edema, nephropathy and neuropathy), age-related macular degeneration, retinal vein occlusion, cerebral edema, ischemia-reperfusion injury, angiogenesis (e.g., in cancer), asthma, anaphylaxis, and cerebrovascular complications of neurological conditions (e.g., Alzheimer's disease, Parkinson's disease, multiple sclerosis, CNS infections, and glioblastoma).

[0039] Plasma kallikrein (PK) is a serine protease that exists in plasma as an inactive zymogen precursor, plasma prekallikrein (pre-PK), and is proteolytically activated by FXIIa. In a positive feedback loop, PK proteolytically activates the zymogen FXII, further forming FXIIa and amplifying its own activation. FXIIa also activates the zymogen FXI to active FXIa, thereby initiating the intrinsic (contact) pathway of blood coagulation, generating thrombin, and cleaving fibrinogen. Importantly, PK cleaves high-molecular-weight kininogen (HMWK) to produce bradykinin. Bradykinin can open the tight junctions between endothelial cells that line blood vessels by activating the bradykinin receptors B1 and B2 present on the surface of endothelial cells, thus allowing fluids and plasma proteins, a condition known as increased vascular permeability, to extravasate from the blood vessels into the tissues. The breakdown of the tight junctions of the blood-brain barrier and the subsequent leakage of plasma and proteins into the brain (edema) have also been associated with Alzheimer's disease, Parkinson's disease, and multiple sclerosis (MS), as well as CNS infections and brain tumors. For example, peritumoral brain edema causes poor prognosis in patients with glioblastoma (K. Schoenegger et al., Eur J Neurol. (2009) 16(7):874-78). When vascular permeability is increased due to the formation of bradykinin, excess amounts of fluid can accumulate in many tissues and organs in various diseases, such as angioedema, cystoid macular edema, diabetic macular edema, macular edema after retinal vein occlusion, cerebral vasogenic edema following stroke or head trauma, and capillary leak syndrome. For example, compound 1 was found to reduce the retinal vascular permeability of angiotensin II-treated rodents, as does the BK receptor antagonist Hoe-140 (J.A. Phipps et al., Hypertension (2009) 53:175-81). It has also been found that the activation of pre-PK and the contact system can cause anaphylaxis in patients treated with contaminated heparin (T.K. Kishimoto et al., N. Engl. J. Med. (2008) 358:2457-67).

[0040] The importance of BK in angioedema is further illustrated in hereditary angioedema, in which individuals have little or no functional C1 inhibitor, the major endogenous inhibitor of PK. When high levels of bradykinin are generated in these patients, fluid and protein leak extravascularly from the plasma into soft tissues, producing severe edema. C1 inhibitor is also known to be involved in the pathogenesis of age-related macular degeneration (S. Ennis et al., Lancet (2008) 372:1828-34) and ischemia-reperfusion injury following organ transplantation or myocardial infarction (D. Inderbitzin et al., Eur. Surg. Res. (2004) 36:142-47; G. Horstick et al., Circulation (2001) 104:3125-31). Bradykinin and its receptors have been found to be involved in tumor angiogenesis (Y. Ikeda et al. Cancer Res (2004) 64:5178-85), pulmonary hypertension (L. Taraseviciene-Stewart et al. Peptides (2005) 26:1292-300) and asthma (P.J. Barnes, “Recent Progress on Kinins”, (1992) AAS38 / III, Birkhauser Verlag, Basel).

[0041] In patients with angioedema, small polypeptide PK inhibitors (DX-88, ecallantide) reduce edema in patients with hereditary angioedema (A. Williams et al., Transfus. Apher. Sci. (2003) 29:255-58; L. Schneider et al., J Allergy Clin Immunol. (2007) 120(2):416-22; J.H. Levy et al., Expert Opin. Invest. Drugs (2006) 15:1077-90). The bradykinin B2 receptor antagonist icatibant is also effective in the treatment of hereditary angioedema (K. Bork et al., J. Allergy Clin. Immunol. (2007) 119:1497-503). Since PK generates bradykinin, inhibition of PK blocks bradykinin production as well.

[0042] In thrombosis due to fibrinolytic therapy (e.g., tissue plasminogen activator, streptokinase), high levels of PK were seen among patients undergoing fibrinolytic therapy (H.M. Hoffmeister et al., J. Cardiovasc. Pharmacol. (1998) 31:764 - 72). Activation mediated by endogenous pathway plasmin was found to occur in plasma and blood and was markedly attenuated in plasma from individuals lacking any of the endogenous pathway components (G.A. Ewald et al., Circulation (1995) 91:28 - 36). Individuals with acute MI had elevated levels of activated PK and thrombin (H.M. Hoffmeister et al., Circulation (1998) 98:2527 - 33).

[0043] Ecarinide decreased cerebral edema, infarct volume, and neurological deficits in an animal model of ischemic stroke (C. Storini et al., J. Pharm. Exp. Ther. (2006) 318:849 - 54). C1 - INH decreased infarct size in a mouse model of middle cerebral artery occlusion (M.G. De Simoni et al., Am. J. Pathol. (2004) 164:1857 - 63; N. Akita et al., Neurosurg. (2003) 52:395 - 400). Compound 1 was found to decrease infarct volume and cerebral vasogenic edema in a rat model of ischemic stroke and to decrease the expansion of intracerebral hemorrhage in a model of hemorrhagic stroke (WO 2009 / 0971 pamphlet). B2 receptor antagonists were found to be neuroprotective with respect to infarct volume, brain swelling, and neutrophil accumulation in an animal model of ischemic stroke (S. Zausinger et al., Acta Neurochir. Suppl. (2003) 86:205 - 07; D.B. Lumenta et al., Brain Res. (2006) 1069:227 - 34; L. Ding - Zhou et al., Br. J. Pharmacol. (2003) 139:1539 - 47).

[0044] In diabetic patients, especially those with proliferative retinopathy, pre-PK levels are found to be higher and to correlate with fructosamine levels (B.-B. Gao et al., Nature Med. (2007) 13:181-88; K. Kedzierska et al., Archives Med. Res. (2005) 36:539-43). Pre-Pk is also elevated in diabetic patients and is found to be highest in patients with sensorimotor neuropathy (M. Christie et al., Thromb. Haemostas. (1984) 52:221-23). Pre-PK levels are elevated in diabetic patients, are associated with increased blood pressure, correlate independently with albumin excretion rate, and are elevated in diabetic patients with macroalbuminuria, suggesting that pre-PK can be a marker of progressive nephropathy (A.A. Jaffa et al., Diabetes (2003) 52:1215-21). B1 receptor antagonists have been found to reduce the increased vascular permeability and plasma leakage into various organs, including the skin and retina, in rats with streptozocin-induced diabetes (S.R. Lawson et al., Eur. J. Pharmacol. (2005) 514:69-78; S.R. Lawson et al., Regul Pept. (2005) 124:221-24). B1 receptor antagonists can also prevent streptozocin-treated mice from developing hyperglycemia and renal insufficiency (A. Zuccollo et al., Can. J. Physiol. Pharmacol. (1996) 74:586-89).

[0045] One aspect of the present invention is a crystalline form of 1-benzyl-N-(4-carbamimidoylbenzyl)-1H-pyrazole-4-carboxamide acetate (Compound 1) that substantially does not contain other polymorphic forms of Compound 1. One embodiment of the present invention is a crystalline form having less than 10% by weight of any other polymorphic form. An embodiment of the present invention is a crystalline form having less than 5% by weight of any other polymorphic form. An embodiment of the present invention is a crystalline form having less than 1% by weight of any other polymorphic form.

[0046] Another aspect of the present invention is an amorphous form of Compound 1 that substantially does not contain other polymorphic forms of Compound 1.

[0047] Another aspect of the present invention is a pharmaceutical composition comprising a pharmaceutically acceptable carrier and Compound 1 in a solid form that substantially does not contain any other solid forms of Compound 1.

[0048] Another aspect of the present invention is a method for producing Compound 1 in a solid form that substantially does not contain any other solid forms of Compound 1.

[0049] Preparation of the Compound The compound of formula I is produced by the process described below.

Chemical formula

[0050] The process of the present invention is optimized to a sufficient extent for commercialization in terms of yield and purity. 1-Benzyl-1H-pyrazole-4-carboxylic acid (“Compound 4”) can be obtained from commercial sources or prepared from commercially available precursor compounds by methods known in the art. For example, 1-benzyl-1H-pyrazole-4-carboxylic acid can be prepared from ethyl 1H-pyrazole-4-carboxylate by contacting it with a suitably substituted benzyl halide (e.g., benzyl bromide) in an aprotic solvent in the presence of a strong base, followed by hydrolysis of the ester. For example, ethyl 1H-pyrazole-3-carboxylate is treated with K2CO3 in acetone, followed by treatment with benzyl bromide to produce the compound ethyl 1-benzyl-1H-pyrazole-3-carboxylate. This ethyl ester is treated, for example, with KOH in methanol to provide the free acid (Compound 4).

[0051] A. Process Step 1: (A) Compound 4 can be coupled with 4-aminomethyl-benzonitrile using 1-propane-phosphonic acid cyclic anhydride (T3P®) and triethylamine (Et3N) in an aprotic solvent to produce 1-benzyl-N-(4-cyanobenzyl)-1H-pyrazole-4-carboxamide (“Compound 3”).

[0052] In one group of process embodiments, T3P® is provided as a 50% ethyl acetate solution. Compound 4 and 4-aminomethyl-benzonitrile may be provided in approximately equimolar amounts, or one reactant may be provided in an excess in the range of about 0.2 to about 5 equivalents compared to the other reactant. In process embodiments, the ratio of Compound 4 to 4-aminomethyl-benzonitrile is about 0.2 to about 5, about 0.5 to about 2, about 0.9 to about 1.2, or about 1.0. T3P® may be provided in a similar range of ratios. In process embodiments, the ratio of T3P® to Compound 4 may be about 0.5 to about 5, about 0.8 to about 4, about 1.0 to about 3, about 1.2 to about 2.0, and about 1.2 to about 1.8. Triethylamine may also be provided in a range of ratios to Compound 4. In process embodiments, the ratio of Et3N to Compound 4 is about 0.5 to about 10, about 1 to about 8, about 2 to about 5, and about 3 to about 5.

[0053] In some embodiments of the process, an aprotic solvent is used, such as, but not limited to, dichloromethane (DCM), tetrahydrofuran (THF), methyl ethyl ketone (MEK), dimethyl sulfoxide (DMSO), ethyl acetate (EtOAc), methyl t-butyl ether (MTBE), and mixtures thereof. In process embodiments, the aprotic solvent is DCM.

[0054] The coupling of Compound 4 and 4-(aminomethyl)benzonitrile is typically carried out in a temperature range of about 0 °C to about 100 °C. In some embodiments, the coupling reaction temperature is about 15 °C to approximately the reflux temperature of the selected aprotic solvent. In process embodiments, the reaction temperature is about 20 °C to about 80 °C. In process embodiments, the reaction temperature is about 20 °C to about 30 °C.

[0055] The above coupling reaction is typically carried out for an appropriate length of time such that the reaction is substantially complete, which may vary with the selected aprotic solvent and reaction temperature. In embodiments of the process, the reaction time is from about 30 minutes to about 48 hours. In embodiments of the process, the reaction time is from about 1 hour to about 24 hours. In other embodiments of the process, the reaction time is from about 4 hours to about 12 hours. In still other embodiments of the process, the reaction time is from about 6 hours to about 10 hours. In some embodiments of the process, the reaction time is about 8 hours.

[0056] In some embodiments of the process, the reaction is carried out under an inert atmosphere or under anhydrous conditions. In some embodiments of the process, the reaction is carried out under a nitrogen atmosphere.

[0057] (B) Compound 3, which is the product of the above coupling reaction, is purified by extraction. Generally, (i) an aprotic solvent containing Compound 3 is combined with water, (ii) mixed well (e.g., by stirring or shaking), (iii) the organic layer and the aqueous layer are separated, (iv) the aqueous layer is removed, and (v) the organic layer is dried to remove water. These steps may be repeated together or individually, one, two, or more than three times. Further, the water may also contain salts such as NaCl, NaHCO3, and the like. The aqueous layer may also be extracted with an organic solvent such as DCM, and the organic solvent can be combined with the other organic layer obtained. This organic layer may then be dried over an appropriate desiccant such as sodium sulfate.

[0058] In some embodiments of the process, Compound 3 in DCM is stirred with water, the layers are separated, then stirred with 10% aqueous NaHCO3, separated, stirred with saturated aqueous NaCl, separated, and then dried over Na2SO4.

[0059] In some embodiments of the process, the dried organic layer is concentrated under reduced pressure, taken up in acetone, washed with water, filtered, suction dried, and then vacuum dried (or dried under reduced pressure) to provide purified Compound 3. The drying step may be carried out at a high temperature, for example, above about 25 °C, above about 30 °C, above about 35 °C, above about 40 °C, above about 45 °C, above about 50 °C, above about 55 °C, and above about 60 °C, etc. The drying temperature is generally below the melting point of Compound 3 and may be below about 150 °C, below about 120 °C, below about 100 °C, below about 90 °C, below about 80 °C, below about 75 °C, below about 70 °C, and below about 65 °C.

[0060] B. Process Step 2: (A) Compound 3 is then contacted with hydroxylamine (NH2OH) or a salt thereof in the presence of a weak base in a suitable solvent to provide a solution of Compound 2. In embodiments of the process, the hydroxylamine is hydroxylamine hydrochloride. Hydroxylamine or a hydroxylamine salt is added to the reactants in a ratio of about 10 to about 0.5 with respect to Compound 3. In some embodiments of the process, the ratio of NH2OH or salt to Compound 3 is about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, about 2, or about 1. In some embodiments of the process, the ratio is at least about 0.5, about 1, about 2, or about 3.

[0061] In some embodiments of the process, the weak base used for the conversion of Compound 3 to Compound 2 is triethylamine or diisopropylamine. The weak base is generally added to the reaction mixture in a ratio of about 10 to about 0.5 with respect to Compound 3. In some embodiments of the process, the ratio of the weak base to Compound 3 is about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, about 2, or about 1. In some embodiments of the process, this ratio is at least about 0.5, about 1, about 2, or about 3.

[0062] In some embodiments of the process (the conversion of Compound 3 to Compound 2), the solvent is ethanol, isopropanol, methanol, DCM, EtOAc, or a mixture thereof.

[0063] In some embodiments of the process for the conversion of Compound 3 to Compound 2, the reaction is carried out at elevated temperatures such as, for example, above about 25°C, above about 30°C, above about 35°C, above about 40°C, above about 45°C, above about 50°C, above about 55°C, above about 60°C, about 65°C, and above about 70°C. The reaction temperature is generally below the reflux temperature of the selected solvent and may be below about 120°C, below about 100°C, below about 90°C, below about 80°C, below about 75°C, below about 70°C, and below about 65°C.

[0064] In some embodiments of the process for the conversion of Compound 3 to Compound 2, the reaction time is generally the length of time required for the reaction to substantially complete, which may vary with the specific reactants, aprotic solvent, and reaction temperature selected. In some embodiments of the process, the reaction time is from about 30 minutes to about 48 hours. In embodiments of the process, the reaction time is from about 1 hour to about 24 hours. In embodiments of the process, the reaction time is from about 4 hours to about 12 hours. In embodiments of the process, the reaction time is from about 6 hours to about 10 hours. In some embodiments of the process, the reaction time is about 7 hours.

[0065] (B) The solution of Compound 2 is then (i) concentrated, (ii) the compound is precipitated by adding water, (iii) the solid is filtered, (iv) washed, and (v) dried to provide purified Compound 2. In some embodiments of the process, (i) the concentration is effected by heating the solution containing Compound 2, reducing the pressure, or both. In some embodiments of the process, the solution is concentrated to a volume of about 20% of the volume of the reaction mixture by heating under reduced pressure. In some embodiments of the process, the solid is dried by suction filtration, drying under reduced pressure, drying at elevated temperature, or a combination thereof. In some embodiments of the process, the solid is first dried by suction filtration and then dried at elevated temperature under reduced pressure to provide purified Compound 2.

[0066] C. Process Step 3: (A) Compound 2 is then subjected to reduction conditions in a protic solvent at a high temperature to provide crude Compound 1. In some embodiments of the process, the reduction conditions include catalytic hydrogenation. In some embodiments of the process, Raney nickel and hydrogen are used for catalytic hydrogenation. In some embodiments of the process, the protic solvent is acetic acid. This process provides Compound 1 as an acetate in some embodiments.

[0067] In some embodiments of the process for converting Compound 2 to Compound 1, a high reaction temperature above about 30 °C, above about 35 °C, above about 40 °C, above about 45 °C, above about 50 °C, above about 55 °C, or above about 60 °C is used. The reaction temperature is generally below the reflux temperature of the solvent and below about 120 °C, below about 100 °C, below about 90 °C, below about 80 °C, below about 75 °C, below about 70 °C, below about 65 °C, below about 60 °C, or below about 55 °C. In some embodiments of the process, the reaction temperature is about 50 °C to 55 °C.

[0068] In some embodiments of the process for converting Compound 2 to Compound 1, a metal catalyst is utilized in catalytic hydrogenation. In some embodiments, the metal catalyst contains nickel. In other embodiments, the metal catalyst contains Raney nickel.

[0069] The amount of catalyst used for the reduction of Compound 2 to Compound 1 may vary depending on the selected catalyst and other reaction conditions. In some embodiments of the process, the amount of Raney nickel used (expressed as mol% based on the amount of Compound 2) is at least about 1 mol%, at least about 5 mol%, at least about 10 mol%, at least about 15 mol%, at least about 20 mol%, at least about 25 mol%, at least about 30 mol%, at least about 35 mol%, at least about 40 mol%, at least about 45 mol%, at least about 50 mol%, or at least about 60 mol%.

[0070] Similarly, the amount of hydrogen used (cm of catalyst 3The kg pressure (displayed as kg pressure per unit area) also varies depending on the amount of catalyst selected and other reaction conditions. In some embodiments of the process, the amount of hydrogen is at least about 1 kg / cm 3 , at least about 2 kg / cm 3 , at least about 3 kg / cm 3 , at least about 4 kg / cm 3 , at least about 5 kg / cm 3 , at least about 6 kg / cm 3 , at least about 7 kg / cm 3 , at least about 8 kg / cm 3 , at least about 9 kg / cm 3 , at least about 10 kg / cm 3 , at least about 11 kg / cm 3 , at least about 12 kg / cm 3 , at least about 15 kg / cm 3 , at least about 20 kg / cm 3 , or at least about 25 kg / cm 3 . In some embodiments, in this reaction, about 20 mol% of Raney nickel and about 10 kg / cm 3 of H2 are utilized.

[0071] The reaction time is generally the length of time required for the reduction to be substantially complete and may vary with the specific conditions selected and the reaction temperature. In some embodiments, the reaction time is at least about 30 minutes, at least about 1 hour, at least about 4 hours, at least about 8 hours, at least about 10 hours, at least about 12 hours, at least about 16 hours, at least about 20 hours, or at least about 24 hours. In other embodiments, the reaction time is less than about 48 hours, less than about 40 hours, less than about 36 hours, less than about 30 hours, less than about 24 hours, less than about 18 hours, or less than about 14 hours. In some embodiments, the reaction time is about 12 hours.

[0072] (B) The crude compound 1 reaction mixture is (i) filtered, (ii) the residue is washed with a first solvent, (iii) concentrated to about 10-20% of the reaction mixture volume, and (iv) the reaction mixture is poured into a second solvent in which the compound dissolves poorly, (v) filtered, and (vi) dried to provide a semi-purified product. The first solvent (step ii) may be a lower alkyl alcohol, dimethyl sulfoxide (DMSO), or dimethylformamide (DMF). In some embodiments, the first solvent is methanol or ethanol. In some embodiments of the process, the second solvent (step iv) is ethyl acetate.

[0073] The filtration process (steps i and v) may include suction filtration and may further include washing the solid with an additional second solvent. In embodiments of the process, the drying process (step vi) may include suction drying, drying under reduced pressure, drying at a high temperature, or a combination thereof. In other embodiments of the process, the drying process may include suction drying followed by drying under reduced pressure at a temperature of at least 35°C. In some embodiments, the reduced pressure is less than 600 mmHg, less than 500 mmHg, less than 400 mmHg, less than 300 mmHg, or less than 200 mmHg. In some embodiments, the drying temperature is at least about 40°C, at least about 45°C, or at least about 50°C. One of ordinary skill in the art will understand that the drying temperature is less than either the melting point or the decomposition temperature of compound 1. In some embodiments, the drying temperature is less than about 120°C, less than about 110°C, less than about 100°C, less than about 90°C, less than about 80°C, less than about 70°C, or less than about 65°C.

[0074] (C) The semi - purified product (Compound 1) at this point may still contain unacceptable amounts of nickel (or other catalytic metals). To further purify the product, the dried solid is (i) treated with water, (ii) heated at a high temperature, stirred to form a slurry, (iii) cooled, (iv) filtered, (v) dried for a first time, (vi) incorporated into a mixture of ethanol and acetic acid, (vii) heated to a maintenance temperature, (viii) cooled, (ix) filtered, and (x) dried for a second time to provide a product depleted in nickel. In some embodiments of the process, the high temperature in step (ii) is at least about 30°C, at least about 35°C, at least about 40°C, at least about 45°C, at least about 50°C, at least about 55°C, at least about 60°C, or at least about 65°C. In some embodiments of the process, the high temperature is less than about 100°C, less than about 90°C, less than about 80°C, less than about 70°C, or less than about 65°C. In some embodiments of the process, the high temperature is about 55°C. In some embodiments of the process, the filtering step in step (iv) includes suction filtering and washing with water. In some embodiments of the process, the drying step in step (v) includes suction drying and then drying under reduced pressure at a temperature of at least about 35°C. In some embodiments of the process, the reduced pressure is less than 600 mmHg, less than 500 mmHg, less than 400 mmHg, less than 300 mmHg, or less than 200 mmHg. In some embodiments of the process, the drying temperature in step (v) is at least about 40°C, at least about 45°C, or at least about 50°C. In an embodiment of the process, the drying temperature is less than about 120°C, less than about 110°C, less than about 100°C, less than about 90°C, less than about 80°C, less than about 70°C, or less than about 65°C. In some embodiments of the process, the drying temperature in step (v) is about 45°C.

[0075] In step (vi), the ratio of ethanol to acetic acid may range from about 1:20 to about 20:1. In some embodiments of the process, the ratio is about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, or about 10:1, v / v ethanol:acetic acid.

[0076] In some embodiments of the process, the holding temperature in step (vii) is at least about 30 °C, at least about 35 °C, at least about 40 °C, at least about 45 °C, at least about 50 °C, at least about 55 °C, at least about 60 °C, or at least about 65 °C. In some embodiments of the process, the holding temperature is approximately the reflux temperature of the solvent mixture of ethanol and acetic acid, or less than about 80 °C, less than about 75 °C, less than about 70 °C, less than about 65 °C, or less than about 60 °C. In some embodiments of the process, the holding temperature is approximately the reflux temperature of this solvent mixture.

[0077] In some embodiments of the process, the mixture is maintained at the holding temperature for a time of at least about 10 minutes, at least about 20 minutes, at least about 30 minutes, at least about 45 minutes, at least about 60 minutes, at least about 90 minutes, at least about 120 minutes, at least about 150 minutes, at least about 240 minutes, or at least about 3 hours. In some embodiments of the process, this time is less than or equal to about 5 hours, less than or equal to about 4 hours, less than or equal to about 3 hours, less than or equal to about 2 hours, less than or equal to about 1 hour, less than or equal to about 30 minutes. In some embodiments of the process, this time is about 1 hour.

[0078] The cooling described in step (viii) is generally carried out over a time period of at least about 10 minutes, at least about 20 minutes, at least about 30 minutes, at least about 45 minutes, at least about 1 hour, or at least about 2 hours. The final temperature of step (viii) is less than about 35 °C, less than about 30 °C, less than about 25 °C, less than about 20 °C, less than about 15 °C, less than about 10 °C, or less than about 5 °C. In some embodiments of the process, the final temperature of step (viii) is approximately ambient temperature. In some embodiments, step (viii) further includes stirring the mixture.

[0079] The drying in step (ix) may further include washing with a lower alkyl alcohol. In some embodiments, the filtered solid is washed with ethanol. The drying step of step (x) may include a step of suction drying, a step of drying under reduced pressure, a step of drying at a high temperature, or a combination thereof. In some embodiments of the process, the drying process of step (x) includes suction drying, followed by drying under reduced pressure at a temperature of at least 35 °C. In an embodiment of the process, the reduced pressure is less than 600 mmHg, less than 500 mmHg, less than 400 mmHg, less than 300 mmHg, or less than 200 mmHg. In some embodiments of the process, the drying temperature of step (x) is at least about 40 °C, at least about 45 °C, or at least about 50 °C. As described above, the drying temperature used is less than the melting point of Compound 1 and less than the decomposition temperature of Compound 1. In some embodiments of the process, the drying temperature is less than about 120 °C, less than about 110 °C, less than about 100 °C, less than about 90 °C, less than about 80 °C, less than about 70 °C, or less than about 65 °C. In some embodiments of the process, the drying temperature of step (x) is about 45 °C. In some embodiments of the process, steps (vi) to (x) are repeated 1, 2, or 3 times. In an embodiment of the process, steps (vi) to (x) are repeated once.

[0080] Crystal form Aspects of the present invention are crystalline forms of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate (Compound 1) that substantially do not contain other polymorphic forms of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate. Embodiments of the present invention are forms having less than 10% by weight of any other polymorphic form. Embodiments of the present invention are forms having less than 5% by weight of any other polymorphic form. Embodiments of the present invention are forms having less than 1% by weight of any other polymorphic form.

[0081] A. Form I: (A) The nickel-depleted product of Process Step 3 above is further purified by (i) contacting the compound with a first solvent, (ii) raising the mixture to a first elevated temperature, (iii) adding a second solvent, (iv) cooling the resulting mixture to the crystallization temperature, (v) stirring the mixture, (vi) filtering the solid, and (vii) drying the solid to provide Compound 1 as pure anhydrous crystalline Form I.

[0082] In some embodiments, the first solvent in step (i) is methanol, ethanol, 1-propanol, or 2-propanol, or a mixture thereof. In some embodiments, the lower alkyl alcohol is methanol.

[0083] In some embodiments, the first high temperature of step (ii) is at least about 30°C, at least about 35°C, at least about 40°C, at least about 45°C, at least about 50°C, at least about 55°C, at least about 60°C, or at least about 65°C. The high temperature is below the reflux temperature of the first solvent, or below about 80°C, below about 75°C, below about 70°C, below about 60°C, below about 55°C, below about 50°C, below about 45°C, below about 40°C, or below about 35°C. In some embodiments, the high temperature is about 55°C. In some embodiments, step (ii) further includes maintaining the mixture at or near the high temperature until Compound 1 is completely dissolved and a clear solution is formed. In some embodiments, step (ii) further includes slowly cooling the solution to a second high temperature. In some embodiments, the second high temperature is about 5°C, about 10°C, about 15°C, or about 20°C lower than the first high temperature. The second high temperature is about 5°C, about 10°C, about 15°C, or about 20°C higher than 20°C. In some embodiments, step (ii) also includes filtering the solution.

[0084] In some embodiments, the second solvent in step (iii) is MTBE or THF. In some embodiments, the second solvent is MTBE. In some embodiments, the first solvent and the second solvent are anhydrous. In some embodiments, the second solvent is added slowly over a long period of at least about 20 minutes, at least about 30 minutes, at least about 45 minutes, at least about 60 minutes, at least about 75 minutes, at least about 90 minutes, at least about 105 minutes, at least about 120 minutes, at least about 150 minutes, at least about 180 minutes, or at least about 240 minutes. The long period is less than about 24 hours, less than about 18 hours, less than about 12 hours, less than about 8 hours, less than about 6 hours, less than about 5 hours, less than about 4 hours, less than about 3 hours, less than about 2 hours, or less than about 1 hour. In some embodiments, the long period is about 2 hours. The ratio of the first solvent to the second solvent may vary from about 1:20 to about 20:1, v / v. In some embodiments, the ratio of MeOH to MTBE is about 5:1, about 4:1, about 3:1, about 2.7:1, about 2.5:1, about 2.3:1, about 2:1, about 1.5:1, about 1.3:1, about 1.2:1, about 1:1, about 1:1.5, about 1:2, about 1:3, or about 1:4.

[0085] In some embodiments, step (iii) further includes adding seed crystals.

[0086] The crystallization temperature in step (iv) is about 35°C or less, less than about 30°C, less than about 25°C, less than about 20°C, less than about 15°C, less than about 10°C, or about 5°C. In some embodiments, the crystallization temperature is about 25°C. One of ordinary skill in the art will understand that cooling generally occurs over a long period. In some embodiments, the cooling time in step (iv) is at least about 20 minutes, at least about 30 minutes, at least about 45 minutes, at least about 60 minutes, at least about 75 minutes, at least about 90 minutes, at least about 105 minutes, at least about 120 minutes, at least about 150 minutes, or at least about 180 minutes. In some embodiments, the cooling time is from about 45 minutes to about 90 minutes.

[0087] In some embodiments of the present invention, step (v) further includes adding an additional amount of the second solvent over a long period of time. In some embodiments, the long period of time in step (v) is at least about 20 minutes, at least about 30 minutes, at least about 45 minutes, at least about 60 minutes, at least about 75 minutes, at least about 90 minutes, at least about 105 minutes, at least about 120 minutes, or at least about 150 minutes. In other embodiments, the long period of time is less than about 24 hours, less than about 18 hours, less than about 12 hours, less than about 8 hours, less than about 6 hours, less than about 5 hours, less than about 4 hours, less than about 3 hours, less than about 2 hours, or less than about 1 hour.

[0088] The filtering step of step (vi) can further include washing the solid with an additional amount of the second solvent. The drying step of step (vii) may include suction drying, drying under reduced pressure, drying at a high temperature, or a combination thereof. In some embodiments, the drying of the invention in step (x) includes suction drying and subsequently drying under reduced pressure at a temperature of at least 35°C. In some embodiments, the reduced pressure is less than 600 mmHg, less than 500 mmHg, less than 400 mmHg, less than 300 mmHg, or less than 200 mmHg. In some embodiments, the drying temperature in step (vii) is at least about 40°C, at least about 45°C, or at least about 50°C. As described above, the drying temperature is less than the melting point of Compound 1 and less than the decomposition temperature of Compound 1. In some embodiments, the drying temperature is less than about 120°C, less than about 110°C, less than about 100°C, less than about 90°C, less than about 80°C, less than about 70°C, or less than about 65°C. In some embodiments, the drying temperature in step (vii) is about 45°C.

[0089] Aspects of the present invention are the crystalline form of Compound 1 ( "Form I") which is anhydrous, has a melting point of about 253 ° C, and a water solubility of about 8.3 mg / mL at 25 ° C. Embodiments of the present invention are that Form I is a crystalline form of Compound 1 having an XRPD pattern that includes peaks at 10.0, 18.1, 18.6, 20.1, and 23.9 degrees ± 0.5 degrees, ± 0.2 degrees, or ± 0.1 degrees, 2θ, and this XRPD pattern is made using CuKα radiation. Embodiments of the present invention are the crystalline form of Compound 1, wherein the XRPD pattern further includes peaks at 20.5 degrees ± 0.5 degrees, ± 0.2 degrees, or ± 0.1 degrees, 2θ. Embodiments of the present invention are the crystalline form of Compound 1 having an XRPD pattern that is substantially similar to the pattern of Figure 1.

[0090] Aspects of the present invention are methods of producing Form I by contacting a solid form of Compound 1 with a solvent selected from the group consisting of methanol, isopropanol, tetrahydrofuran, 2-methyl-tetrahydrofuran, anhydrous acetone, dichloromethane, diethyl ether, 3-methyl-1-butanol, and nitromethane, or mixtures thereof, for a time sufficient to produce Form I. Embodiments of the present invention are methods wherein the solvent is methanol.

[0091] B. Form II: Compound 1 in any solid form is converted to Form II by the following process. Compound 1 is contacted with (i) a mixture of an organic solvent and water, (ii) heated for a first time and maintained, (iii) a second solvent is added, (iv) the mixture is cooled and maintained for a second time to crystallize, and (v) the resulting solid is separated to provide Compound 1 as pure crystalline Form II. In some embodiments, the organic solvent in step (i) is methanol, isopropanol, THF, or acetone. In some embodiments, the organic solvent is methanol. In some embodiments, the mixture of organic solvent and water contains at least about 1% water, at least about 2% water, at least about 3% water, at least about 4% water, or at least about 5% water. In some embodiments, the mixture of organic solvent and water contains at least about 10% water, at least about 15% water, at least about 20% water, at least about 25% water, at least about 30% water, at least about 35% water, at least about 40% water, at least about 45% water, or at least about 50% water. In some embodiments, the mixture of organic solvent and water contains up to about 45% water, up to about 40% water, up to about 35% water, up to about 30% water, up to about 25% water, up to about 20% water, up to about 15% water, or up to about 10% water.

[0092] The high temperature in step (ii) is at least about 30°C, at least about 35°C, at least about 40°C, at least about 45°C, at least about 50°C, at least about 55°C, at least about 60°C, or at least about 65°C. The high temperature is below the reflux temperature of the solvent mixture or below about 80°C, below about 75°C, below about 70°C, below about 60°C, below about 55°C, below about 50°C, below about 45°C, below about 40°C, or below about 35°C. In some embodiments, the high temperature is about 55°C. In some embodiments, step (ii) further includes maintaining the mixture at or near the high temperature until Compound 1 is completely dissolved and a clear solution is formed. In some embodiments, step (ii) also includes filtering this solution. In some embodiments, the high temperature is about 55°C. In some embodiments, step (ii) further includes maintaining the mixture at or near the high temperature until Compound 1 is completely dissolved and a clear solution is formed. In some embodiments, this time may be about 1 minute, about 2 minutes, about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 45 minutes, about 60 minutes, or about 120 minutes. In some embodiments, this time is less than about 180 minutes, less than about 120 minutes, less than about 60 minutes, less than about 45 minutes, or less than about 30 minutes.

[0093] The second solvent in step (iii) is MTBE or THF. In some embodiments, the second solvent is MTBE. Step (iii) may optionally include adding seed crystals of Form II either before or after the addition of the second solvent.

[0094] The crystallization temperature in step (iv) is below about 35°C, less than about 30°C, less than about 25°C, less than about 20°C, less than about 15°C, less than about 10°C, or less than about 5°C. In some embodiments, the crystallization temperature is about 25°C.

[0095] The cooling in step (iv) generally occurs over an extended period of time. In some embodiments, the cooling time of step (iv) is at least about 20 minutes, at least about 30 minutes, at least about 45 minutes, at least about 60 minutes, at least about 75 minutes, at least about 90 minutes, at least about 105 minutes, at least about 120 minutes, at least about 150 minutes, or at least about 180 minutes. In some embodiments, this cooling time is from about 45 minutes to about 90 minutes. In some embodiments, the crystallization time / maintenance time of step (iv) is at least about 20 minutes, at least about 30 minutes, at least about 45 minutes, at least about 60 minutes, at least about 75 minutes, at least about 90 minutes, at least about 105 minutes, at least about 120 minutes, or at least about 150 minutes. In some embodiments, this time is less than about 24 hours, less than about 18 hours, less than about 12 hours, less than about 8 hours, less than about 6 hours, less than about 5 hours, less than about 4 hours, less than about 3 hours, less than about 2 hours, or about 1 hour.

[0096] The separating step of step (v) is substantially similar to steps (vi) and (vii) of the process of Form I.

[0097] Aspects of the present invention are crystalline forms of Compound 1 (``Form II'') that are hydrated and characterized by a melting point of about 253 °C and a water solubility of about 5.5 mg / mL at 25 °C. Embodiments of the present invention are crystalline forms of Compound 1 having an XRPD pattern that includes peaks at 4.2, 5.6, 8.4, 17.8, and 19.8 degrees ±0.5 degrees, ±0.2 degrees, or ±0.1 degrees, 2θ, where this XRPD pattern is made using CuKα radiation. Embodiments of the present invention are crystalline forms of Compound 1 where this XRPD pattern further includes a peak at 12.7 degrees ±0.5 degrees, ±0.2 degrees, or ±0.1 degrees, 2θ. Embodiments of the present invention are crystalline forms of Compound 1 having an XRPD pattern that is substantially similar to the pattern of Figure 2.

[0098] Aspects of the present invention include contacting a solid form of Compound 1 with a mixture of water and a solvent selected from isopropanol, tetrahydrofuran, acetone, and ethyl acetate, or combinations thereof, for a time sufficient to produce Form II. In one embodiment, the solution is ethyl acetate. In another embodiment, the solution is methanol. In another embodiment, the method further includes adding MIBK to the solution.

[0099] C. Form III: (A) Compound 1 in any solid form is converted to Form III by the following process. Compound 1 is (i) contacted with an organic solvent, (ii) heated and maintained for a first time, (iii) a second solvent is added, (iv) the mixture is cooled and maintained for a second time to crystallize, and (v) the resulting solid is separated to provide Compound 1 as pure crystalline Form III.

[0100] In some embodiments, the organic solvent in step (i) is MeOH. In some embodiments, the organic solvent contains less than about 10% water. In some embodiments, the organic solvent contains about 10% or less water, about 8% or less water, about 5% or less water, about 2% or less water, about 1% or less water, or about 0.1% or less water.

[0101] The high temperature in step (ii) is at least about 30 °C, at least about 35 °C, at least about 40 °C, at least about 45 °C, at least about 50 °C, at least about 55 °C, at least about 60 °C, or at least about 65 °C. The high temperature is below the reflux temperature of the solvent mixture, or below about 80 °C, below about 75 °C, below about 70 °C, below about 60 °C, below about 55 °C, below about 50 °C, below about 45 °C, below about 40 °C, or below about 35 °C. In some embodiments, the high temperature is about 55 °C. In some embodiments, step (ii) further includes maintaining the mixture at or near the high temperature until compound 1 is completely dissolved and a clear solution is formed. In some embodiments, step (ii) also includes filtering this solution. In some embodiments, the high temperature is about 55 °C. In some embodiments, step (ii) further includes maintaining the mixture at or near the high temperature until compound 1 is completely dissolved and a clear solution is formed. In some embodiments, this time may be about 1 minute, about 2 minutes, about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 45 minutes, about 60 minutes, or about 120 minutes. In some embodiments, this time is less than about 180 minutes, less than about 120 minutes, less than about 60 minutes, less than about 45 minutes, or less than about 30 minutes.

[0102] In some embodiments, the second solvent in step (iii) is acetonitrile (CH3CN). The ratio of acetonitrile to the MeOH solution is at least about 1:1, at least about 1:2, at least about 1:3, at least about 1:4, at least about 1:5, at least about 1:7, at least about 1:10, at least about 1:20, at least about 1:30, at least about 1:40, or at least about 1:50. Step (iii) may optionally include adding seed crystals of Form III either before or after the addition of the second solvent.

[0103] The crystallization temperature in step (iv) is about 35 °C or lower, less than about 30 °C, less than about 25 °C, less than about 20 °C, less than about 15 °C, less than about 10 °C, or less than about 5 °C. In some embodiments, the crystallization temperature is about 25 °C. Cooling occurs over a long period of time. In some embodiments, the cooling time in step (iv) is at least about 20 minutes, at least about 30 minutes, at least about 45 minutes, at least about 60 minutes, at least about 75 minutes, at least about 90 minutes, at least about 105 minutes, at least about 120 minutes, at least about 150 minutes, or at least about 180 minutes. In some embodiments, the cooling time is from about 45 minutes to about 90 minutes. In some embodiments, the crystallization time / maintenance time of step iv is at least about 20 minutes, at least about 30 minutes, at least about 45 minutes, at least about 60 minutes, at least about 75 minutes, at least about 90 minutes, at least about 105 minutes, at least about 120 minutes, or at least about 150 minutes. In some embodiments, the time is less than about 24 hours, less than about 18 hours, less than about 12 hours, less than about 8 hours, less than about 6 hours, less than about 5 hours, less than about 4 hours, less than about 3 hours, less than about 2 hours, or less than about 1 hour.

[0104] The separating step in step (v) is substantially similar to steps (vi) and (vii) of the process of Form I.

[0105] (B) Form III can also be produced by an alternative process. Compound 1 is contacted with (i) an organic solvent, (ii) heated and maintained for a first time, (iii) the solvent is rapidly removed under reduced pressure, and (iv) the resulting solid is separated to provide Compound 1 as pure crystalline Form III. In some embodiments, the organic solvent in step (i) is MeOH, or a mixture of MeOH and DCM. In some embodiments, the organic solvent is MeOH. In some embodiments, the organic solvent is a mixture of MeOH and DCM. In some embodiments, the ratio of MeOH to DCM is about 1:1. In some embodiments, the organic solvent contains less than about 10% water. In some embodiments, the organic solvent contains about 10% or less water, about 8% or less water, about 5% or less water, about 2% or less water, about 1% or less water, or about 0.1% or less water.

[0106] The high temperature in step (ii) is at least about 30°C, at least about 35°C, at least about 40°C, at least about 45°C, at least about 50°C, at least about 55°C, at least about 60°C, or at least about 65°C. The high temperature is below the reflux temperature of the solvent mixture, or below about 80°C, about 75°C, about 70°C, about 60°C, about 55°C, about 50°C, about 45°C, about 40°C, or about 35°C. In some embodiments, the high temperature is about 55°C. In some embodiments, step ii further includes maintaining the mixture at or near the high temperature until Compound 1 is completely dissolved and a clear solution is formed. In some embodiments, step (ii) also includes filtering the solution. In some embodiments, the high temperature is about 55°C. In some embodiments, this time may be about 1 minute, about 2 minutes, about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 45 minutes, about 60 minutes, or about 120 minutes. In some embodiments, this time is less than about 180 minutes, less than about 120 minutes, less than about 60 minutes, less than about 45 minutes, or less than about 30 minutes.

[0107] The solvent removal in step (iii) is carried out rapidly under reduced pressure. This pressure may be less than about 700 mmHg, less than about 600 mmHg, less than about 500 mmHg, less than about 400 mmHg, less than about 350 mmHg, less than about 300 mmHg, less than about 250 mmHg, less than about 200 mmHg, less than about 150 mmHg, less than about 100 mmHg, or less than about 50 mmHg. The rapid solvent removal is carried out in less than about 30 minutes, less than about 20 minutes, less than about 15 minutes, less than about 10 minutes, less than about 9 minutes, less than about 8 minutes, less than about 7 minutes, less than about 6 minutes, less than about 5 minutes, less than about 4 minutes, less than about 3 minutes, less than about 2 minutes, or less than about 1 minute.

[0108] The separating step of step (iv) is substantially similar to steps (vi) and (vii) of the process of Form I.

[0109] Aspects of the present invention are crystalline forms of Compound 1 ( "Form III") characterized by not being an anhydrate, having a melting point of about 249 ° C, and a water solubility of about 7.2 mg / mL at 25 ° C. Embodiments of the present invention are crystalline forms of Compound 1 having an XRPD containing peaks at 7.7, 10.7, 20.1, 23.7 and 24.3 degrees ± 0.5 degrees, ± 0.2 degrees, or ± 0.1 degrees, 2θ, this XRPD pattern being that of a crystalline form of Compound 1 made using CuKα radiation. Embodiments of the present invention are crystalline forms of Compound 1 in which this XRPD pattern further includes peaks at 18.5 and 26.5 degrees ± 0.5 degrees, ± 0.2 degrees, or ± 0.1 degrees, 2θ. Embodiments of the present invention are crystalline forms of Compound 1 having an XRPD pattern substantially similar to the pattern of Figure 3.

[0110] Aspects of the present invention are methods of producing Form III by contacting Compound 1 in any solid form with a solvent containing acetonitrile for a time sufficient to produce Form III. Embodiments of the present invention are methods in which this solvent contains less than about 10% water. Embodiments of the present invention are methods in which this solvent contains less than about 5% water. Embodiments of the present invention are methods in which this solvent contains less than about 1% water.

[0111] D. Form IV: (A) Any solid form of Compound 1 is converted to Form IV by the following process. Compound 1 is contacted with (i) an organic solvent, (ii) heated and maintained for a first time, (iii) a second solvent is added, (iv) the mixture is cooled and maintained for a second time to crystallize, and (v) the resulting solid is separated to provide Compound 1 as pure crystalline Form IV.

[0112] In some embodiments, the organic solvent in step (i) is MeOH. In some embodiments, the organic solvent contains less than about 5% water, less than about 2% water, less than about 1% water, or less than about 0.1% water.

[0113] The high temperature in step (ii) is at least about 30°C, at least about 35°C, at least about 40°C, at least about 45°C, at least about 50°C, at least about 55°C, at least about 60°C, or at least about 65°C. The high temperature is below the reflux temperature of this solvent mixture, or below about 80°C, below about 75°C, below about 70°C, below about 60°C, below about 55°C, below about 50°C, below about 45°C, below about 40°C, or below about 35°C. In some embodiments, the high temperature is about 55°C. In some embodiments, step (ii) further includes maintaining the mixture at or near the high temperature until Compound 1 is completely dissolved and a clear solution is formed. In some embodiments, step (ii) also includes filtering this solution. In some embodiments, the high temperature is about 55°C. In some embodiments, step (ii) further includes maintaining the mixture at or near the high temperature until Compound 1 is completely dissolved and a clear solution is formed. In some embodiments, this time may be about 1 minute, about 2 minutes, about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 45 minutes, about 60 minutes, or about 120 minutes. In some embodiments, this time is less than about 180 minutes, less than about 120 minutes, less than about 60 minutes, less than about 45 minutes, or less than about 30 minutes.

[0114] The second solvent in step (iii) is ethyl acetate (EtOAc), isopropyl acetate, MEK, or MIBK. In some embodiments, the second solvent is ethyl acetate (EtOAc), isopropyl acetate, MEK, or MIBK. In some embodiments, the second solvent is EtOAc. In some embodiments, the second solvent is MEK. In some embodiments, the second solvent is MIBK.

[0115] The ratio of the second solvent (in step (i)) to the MeOH solution is at least 0.1:1, at least about 0.2:1, at least about 0.3:1, at least about 0.4:1, at least about 0.5:1, at least about 1:1, at least about 1:2, at least about 1:3, at least about 1:4, at least about 1:5, at least about 1:7, at least about 1:8, at least about 1:9, at least about 1:10, at least about 1:20, at least about 1:30, at least about 1:40, or at least about 1:50.

[0116] Step (iii) may optionally include the step of adding seed crystals of Form IV either before or after the addition of the second solvent.

[0117] The crystallization temperature in step (iv) is about 35 °C or less, less than about 30 °C, less than about 25 °C, less than about 20 °C, less than about 15 °C, less than about 10 °C, or less than about 5 °C. In some embodiments, the crystallization temperature is about 25 °C. The cooling occurs over a long period of time. In some embodiments, the cooling time in step (iv) is at least about 20 minutes, at least about 30 minutes, at least about 45 minutes, at least about 60 minutes, at least about 75 minutes, at least about 90 minutes, at least about 105 minutes, at least about 120 minutes, at least about 150 minutes, or at least about 180 minutes. In some embodiments, the cooling time is from about 45 minutes to about 90 minutes. In some embodiments, the crystallization time / maintenance time in step (iv) is at least about 20 minutes, at least about 30 minutes, at least about 45 minutes, at least about 60 minutes, at least about 75 minutes, at least about 90 minutes, at least about 105 minutes, at least about 120 minutes, or at least about 150 minutes. This time is less than about 72 hours, less than about 48 hours, less than about 24 hours, less than about 18 hours, less than about 12 hours, less than about 8 hours, less than about 6 hours, less than about 5 hours, less than about 4 hours, less than about 3 hours, less than about 2 hours, or less than about 1 hour.

[0118] The separating step in step (v) is substantially similar to steps (vi) and (vii) of the process of Form I.

[0119] Aspects of the present invention are crystalline forms of Compound 1 characterized by having a melting point of about 251 °C (“Form IV”). Embodiments of the present invention are crystalline forms of Compound 1 having an XRPD pattern that includes peaks at 18.5, 19.6 and 23.7 degrees ±0.5 degrees, ±0.2 degrees, or ±0.1 degrees, 2θ, where this XRPD pattern is made using CuKα radiation. Embodiments of the present invention are crystalline forms of Compound 1 where this XRPD pattern further includes peaks at 24.6 degrees ±0.5 degrees, ±0.2 degrees, or ±0.1 degrees, 2θ. Embodiments of the present invention are crystalline forms of Compound 1 having an XRPD pattern that is substantially similar to the pattern of FIG. 4.

[0120] E. Form V - Amorphous Any solid form of Compound 1 is converted to Form V by the following process. Compound 1 is contacted with (i) an aqueous solvent, (ii) heated and maintained for a first time, (iii) this solution is frozen, freeze-dried, and (iv) the resulting solid is separated to provide Compound 1 as pure amorphous Form V.

[0121] In some embodiments, the aqueous solvent in step (i) is water, deionized water, or water for injection. In an embodiment of the present invention, this aqueous solvent is water for injection. In an embodiment of the present invention, this aqueous solvent is deionized water.

[0122] The high temperature in step (ii) is at least about 30°C, at least about 35°C, at least about 40°C, at least about 45°C, at least about 50°C, at least about 55°C, at least about 60°C, or at least about 65°C. This high temperature is below the reflux temperature of this solvent mixture, or below about 80°C, about 75°C, about 70°C, about 60°C, about 55°C, about 50°C, about 45°C, about 40°C, or about 35°C. In an embodiment of the present invention, the high temperature is about 45°C. In some embodiments, step (ii) further includes maintaining this mixture at or near the high temperature until Compound 1 is completely dissolved and a clear solution is formed. In some embodiments, step (ii) also includes filtering this solution. In some embodiments, the high temperature is about 55°C. In some embodiments, step (ii) further includes maintaining this mixture at or near the high temperature until Compound 1 is completely dissolved and a clear solution is formed. In some embodiments, this time may be about 1 minute, about 2 minutes, about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 45 minutes, about 60 minutes, or about 120 minutes. In some embodiments, this time is less than about 180 minutes, less than about 120 minutes, less than about 60 minutes, less than about 45 minutes, or less than about 30 minutes.

[0123] The lyophilization step (iii) occurs over an extended period of time. In some embodiments, the lyophilization time of step (iii) is at least about 20 minutes, at least about 30 minutes, at least about 45 minutes, at least about 60 minutes, at least about 75 minutes, at least about 90 minutes, at least about 105 minutes, at least about 120 minutes, at least about 150 minutes, at least about 180 minutes, at least about 3 hours, at least about 4 hours, at least about 5 hours, at least about 6 hours, at least about 7 hours, at least about 8 hours, at least about 9 hours, or at least about 10 hours. This time is less than about 24 hours, less than about 18 hours, less than about 12 hours, less than about 8 hours, less than about 6 hours, less than about 5 hours, less than about 4 hours, less than about 3 hours, less than about 2 hours, or less than about 1 hour.

[0124] The separation step of step (iv) generally only requires the step of recovering the amorphous solid from the lyophilization apparatus.

[0125] Aspects of the present invention are the amorphous form of Compound 1 ( "Form V") characterized by a melting point of about 240 ° C and a water solubility of about 7.1 mg / mL at 25 ° C. Embodiments of the present invention are amorphous forms of Compound 1 having an XRPD pattern substantially similar to the pattern of Figure 5.

[0126] Formulation Compound 1 is formulated and administered according to methods known in the art. A pharmaceutical composition containing the active ingredient may be, for example, tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions and self-emulsifying agents as described in US Patent Application No. 2002-0012680, hard or soft capsules, syrups, elixirs, solutions, buccal patches, oral gels, chewing gums, chewable tablets, effervescent powders and effervescent tablets, and may be in a form suitable for oral use. Compositions intended for oral use can be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such compositions can contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents, antioxidants and preservatives to obtain a pharmaceutically refined and palatable preparation. Tablets contain the active ingredient mixed with non-toxic pharmaceutically acceptable additives suitable for the manufacture of tablets. These additives can be inert diluents such as cellulose, silicon dioxide, aluminum oxide, calcium carbonate, sodium carbonate, glucose, mannitol, sorbitol, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents such as corn starch or alginic acid; binders such as PVP, cellulose, PEG, starch, gelatin or gum arabic; and lubricants such as magnesium stearate, stearic acid or talc. These tablets may or may not be coated, or may be coated enterically or in another manner by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a longer-lasting effect. For example, time-delay materials such as glyceryl monostearate or glyceryl distearate may be used. They can also be coated by the techniques described in US Patents Nos. 4,256,108, 4,166,452 and 4,265,874 to form osmotic therapeutic tablets for controlled release.

[0127] Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oily medium such as groundnut oil, liquid paraffin or olive oil. Further, emulsions can be prepared with non-aqueous miscible components such as oil and can be stabilized with surfactants such as mono- and diglycerides, PEG esters and the like.

[0128] An aqueous suspension contains the active material mixed with additives suitable for the manufacture of an aqueous suspension. Such additives are suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone, tragacanth gum and gum arabic, and the dispersing or wetting agent can be a natural phosphatide such as lecithin, or a condensate of alkylene oxide and fatty acid such as polyoxyethylene stearate, or a condensate of ethylene oxide and long-chain aliphatic alcohol such as heptadecaethyleneoxycetanol, or a condensate of ethylene oxide and partial ester derived from fatty acid and hexitol such as polyoxyethylene sorbitol monooleate, or a condensate of ethylene oxide and partial ester derived from fatty acid and hexitol anhydride such as polyoxyethylene sorbitan monooleate. An aqueous suspension may also contain one or more preservatives such as ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents such as sucrose or saccharin.

[0129] The oily suspension can be formulated by suspending the active ingredient in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or a mineral oil such as liquid paraffin. The oily suspension may contain a thickening agent such as beeswax, solid paraffin or cetyl alcohol. In order to obtain a palatable oral preparation, sweeteners and flavoring agents such as those shown above can also be added. These compositions can also be preserved by adding an antioxidant such as ascorbic acid.

[0130] A powder or granule having suitable dispersibility for the preparation of an aqueous suspension by adding water provides an active ingredient mixed with a dispersant or wetting agent, a suspending agent and one or more preservatives. Suitable dispersants or wetting agents, and suspending agents are exemplified by those already described above. Other additives such as sweeteners, flavoring agents and coloring agents may also be present.

[0131] The pharmaceutical composition of the present invention may be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil such as olive oil or peanut oil, or a mineral oil such as liquid paraffin, or a mixture thereof. Suitable emulsifiers may be natural rubbers such as gum arabic or tragacanth gum, natural phosphatides such as soybean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides such as sorbitan monooleate, and condensates of this partial ester with ethylene oxide such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners and flavoring agents.

[0132] Syrups and elixirs can be formulated with sweeteners such as glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain demulcents, preservatives, and flavoring agents and coloring agents. Oral solutions can be prepared, for example, in combination with cyclodextrin, PEG and surfactants.

[0133] The pharmaceutical composition may be in the form of an aqueous or oily suspension for sterile injection. This suspension can be formulated using suitable dispersing or wetting agents and suspending agents as described above according to known techniques. The sterile injectable preparation may be a solution or suspension for sterile injection as a solution in a non-toxic parenterally acceptable diluent or solvent, for example in 1,3-butanediol. Acceptable media and solvents that can be used include water, Ringer's solution, and isotonic sodium chloride solution. Furthermore, sterile fixed oils are also conventionally used as solvents or suspending media. For this purpose, any non-irritating fixed oil containing synthetic monoglycerides or diglycerides can be used. Furthermore, fatty acids such as oleic acid are also used in the preparation of injectables.

[0134] Compound 1 can also be administered in the form of a suppository for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating additive that is solid at room temperature but liquid at rectal temperature and thus melts in the rectum to release the drug. Such materials include cocoa butter and polyethylene glycol. Furthermore, the compound can also be administered via delivery to the eye by means of a solution or ointment. Still further, transdermal delivery of the subject compound can also be achieved by means of an iontophoresis-type patch or the like. For topical use, creams, ointments, jellies, solutions or suspensions containing the compound of the present invention are used. As used herein, topical application shall also include the use of gargles and mouthwashes.

[0135] Compound 1 can also be formulated for attachment in a medical device that can include any of a variety of conventional grafts, stents including stent grafts, catheters, balloons, baskets or other devices that can be placed or permanently implanted into a body cavity. As one particular example, it would be desirable to have a device and method capable of delivering the compound of the present invention to a body region treated by an interventional procedure.

[0136] Compound 1 can be attached inside a medical device such as a stent and reach a treatment site for the treatment of a part of the body. The stent is used as a delivery medium for a therapeutic agent (i.e., a drug). Intravascular stents are generally permanently implanted within coronary or peripheral blood vessels. The stent designs include those of U.S. Patent No. 4,733,655 (Palmaz), U.S. Patent No. 4,800,882 (Gianturco), and U.S. Patent No. 4,886,062 (Wiktor). Such designs include both metal and polymer stents, as well as self-expanding and balloon-expandable stents. Also, for example, as disclosed in U.S. Patent No. 5,102,417 (Palmaz), as well as International Publication No. WO 91 / 12779 pamphlet (Medtronic, Inc.) and International Publication No. WO 90 / 13332 pamphlet (Cedars-Sanai Medical Center), U.S. Patent No. 5,419,760 (Narciso, Jr.) and U.S. Patent No. 5,429,634 (Narciso, Jr.), stents can also be used to deliver drugs at the site of contact with the vascular structure.

[0137] The term "deposited" means that the inhibitor is coated, adsorbed, positioned, or incorporated in another manner into the device by methods known in the art. For example, the inhibitor can be embedded in a polymer material that covers or extends over the medical device and released therefrom ("matrix type"), or surrounded by it and released therethrough ("reservoir type"). In the latter example, one or more methods for producing such materials known in the art can be used to trap the inhibitor within the polymer material or couple it to the polymer material. In other formulations, the inhibitor may be linked to the surface of the medical device without the need for a coating by a separable bond and release over time, and can also be removed by an active mechanical or chemical process, or be in a permanently immobilized form that presents the inhibitor at the implantation site.

[0138] The polymer may be either a biostable polymer or a bioabsorbable polymer, depending on the desired release rate or degree of desired polymer stability. Bioabsorbable polymers that can be used include poly(L-lactic acid), polycaprolactone, polyglycolide (PGA), poly(lactide-co-glycolide) (PLLA / PGA), poly(hydroxybutyrate), poly(hydroxybutyrate-co-valerate), polydioxanone, polyorthoester, polyanhydride, poly(glycolic acid), poly(D-lactic acid), poly(L-lactic acid), poly(D,L-lactic acid), poly(D,L-lactide) (PLA), poly(L-lactide) (PLLA), poly(glycolic acid-co-trimethylene carbonate) (PGA / PTMC), polyethylene oxide (PEO), polydioxanone (PDS), polyphosphoester, polyphosphoester urethane, poly(amino acid), cyanoacrylate, poly(trimethylene carbonate), poly(iminocarbonate), copoly(ether-ester) (e.g., PEO / PLA), polyalkylene oxalate, polyphosphazene, and biomolecules such as fibrin, fibrinogen, cellulose, starch, collagen, and hyaluronic acid, poly ε-caprolactone, polyhydroxybutyric acid, polyorthoester, polyacetal, polydihydropyran, polycyanoacrylate, crosslinkable or amphiphilic block copolymers of hydrogels, and other suitable bioabsorbable polymers known in the art are non-limitingly included.Also, biostable polymers with relatively low chronic tissue reactivity, such as polyurethane, silicone, and polyester, can be used, and if they can dissolve and cure or polymerize on the medical device, polyolefin, polyisobutylene, and ethylene-α-olefin copolymer; acrylic polymer and copolymer, halogenated vinyl polymer and copolymer, such as polyvinyl chloride, etc.; polyvinyl pyrrolidone; polyvinyl ether, such as polyvinyl methyl ether, etc.; polyhalogenated vinylidene, such as polyvinylidene fluoride and polyvinylidene chloride, etc.; polyacrylonitrile, polyvinyl ketone; polyvinyl aromatic, such as polystyrene; polyvinyl ester, such as polyvinyl acetate; copolymer of vinyl monomer with each other and with olefin, such as ethylene-methyl methacrylate copolymer, acrylonitrile-styrene copolymer, ABS resin, and ethylene-vinyl acetate copolymer, etc.; pyran copolymer; polyhydroxy-propyl-methacrylamide-phenol; polyhydroxyethyl-aspartoamide-phenol; polyethylene oxide-polylysine substituted by palmitoyl residue; polyamide, such as nylon 66 and polycaprolactam, etc.; alkyd resin, polycarbonate; polyoxymethylene; polyimide; polyether; epoxy resin, polyurethane; rayon; rayon-triacetate; cellulose, cellulose acetate, cellulose butyrate; cellulose acetate butyrate; cellophane; cellulose nitrate; cellulose propionate; cellulose ether; and other polymers such as carboxymethyl cellulose can also be used.

[0139] The polymer and the semipermeable polymer matrix can also be formed as shaped articles such as valves, stents, tubes, artificial organs, and the like. Typically, the polymer is applied to the surface of the implantable device by spin coating, dipping, or spraying. Other methods known in the art can also be utilized for this purpose. Spraying methods include conventional methods and microdeposition methods using inkjet-type dispensers. Additionally, photolithography or 3D printing can be used to deposit the polymer onto the implantable device in order to place the polymer only on specific portions of the device. This coating of the device provides a uniform layer around the device, thereby improving the diffusion of various analytes through the coating of the device.

[0140] Compound 1 can be formulated for release from the polymer coating into the environment in which the medical device is placed. For example, the compound is released in a controlled manner over a long-term time frame (e.g., several months) using at least one of several well-known techniques involving a polymer carrier or layer for controlling elution. Some of these techniques are described previously in U.S. Patent Application Publication No. 2004 / 0243225, the entire disclosure of which is incorporated herein by reference in its entirety.

[0141] Furthermore, for example, as described in U.S. Patent No. 6,770,729, which is incorporated herein by reference in its entirety, the reaction conditions of the reagent and the polymer composition can be manipulated to control the release of the inhibitor from the polymer coating. For example, the diffusion coefficient of one or more polymer coatings can be adjusted to control the release of the inhibitor from the polymer coating. In one variation of this subject matter, an analyte present in the environment in which the medical device is disposed (e.g., an analyte that promotes the disintegration or hydrolysis of a portion of the polymer) can control the ability of the analyte to access one or more components within the polymer composition (and thereby, for example, regulate the release of the inhibitor from the polymer coating) by controlling the diffusion coefficient of one or more polymer coatings. Another embodiment of the process involves a device having a plurality of polymer coatings, each having a plurality of diffusion coefficients. In such an embodiment of the process, the release of the inhibitor from the polymer coating can be regulated by the plurality of polymer coatings.

[0142] The release of the inhibitor from the polymer coating can be controlled by adjusting one or more properties of the polymer composition, such as the presence of one or more endogenous or exogenous compounds or the pH of the polymer composition. For example, certain polymer compositions can be designed to release the inhibitor in response to a decrease in the pH of the polymer composition. Alternatively, certain polymer compositions can be designed to release the inhibitor in response to the presence of hydrogen peroxide.

[0143] Aspects of the present invention are pharmaceutical compositions for treating plasma kallikrein-dependent diseases or conditions, the pharmaceutical compositions comprising a therapeutically effective amount of a solid form of Compound 1 and a pharmaceutically acceptable carrier, wherein this solid form of Compound 1 is Form I, Form II, Form III, Form IV, or Form V, and substantially does not contain any other polymorphic or amorphous forms of Compound 1. Embodiments of the present invention are pharmaceutical compositions wherein this solid form is Form I. Embodiments of the present invention are pharmaceutical compositions wherein this solid form is Form II. Embodiments of the present invention are pharmaceutical compositions wherein this solid form is Form III. Embodiments of the present invention are pharmaceutical compositions wherein this solid form is Form IV. Embodiments of the present invention are pharmaceutical compositions wherein this solid form is Form V.

[0144] Aspects of the present invention are the use of a crystalline form of Compound 1 for the manufacture of a medicament for the treatment of plasma kallikrein-dependent diseases or conditions, wherein this solid form of Compound 1 substantially does not contain any other polymorphic or amorphous forms of Compound 1. Another aspect of the present invention is the use of a crystalline form of Compound 1 for the manufacture of a medicament for the treatment of plasma kallikrein-dependent diseases or conditions, wherein this solid form of Compound 1 substantially does not contain any other polymorphic or amorphous forms of Compound 1.

[0145] Administration The pharmaceutical compositions of the present invention are administered by any of the accepted methods of administration for therapeutically effective amounts of agents that serve a similar utility. The actual amount of Compound 1 administered depends on a number of factors such as the severity of the disease to be treated, the age and relative health of the subject, the potency of the compound being used, the route and form of administration, and other factors.

[0146] Aspects of the invention are methods of treating a plasma kallikrein-dependent disease or condition by administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition of the invention. Embodiments of the invention are such that the plasma kallikrein-dependent disease or condition is selected from diabetic macular edema, diabetic retinopathy, hereditary angioedema due to C1 inhibitor deficiency, acute liver injury, inflammation, anaphylaxis, chemically sensitized kidney injury, ischemic stroke, hemorrhagic stroke, hypertension, hypertensive vascular complications, retinopathy, nephropathy, cerebral vascular edema, pulmonary hypertension, inflammation, pain, acute myocardial infarction, deep vein thrombosis, complications resulting from fibrinolytic therapy, angina, angioedema, sepsis, arthritis, complications of cardiopulmonary bypass, capillary leak syndrome, inflammatory bowel disease, diabetes, diabetic retinopathy, diabetic macular edema, diabetic nephropathy, diabetic neuropathy, age-related macular degeneration, retinal vein occlusion, cerebral edema, ischemia-reperfusion injury, cancer-related angiogenesis, asthma, anaphylaxis, and cerebral vascular complications of Alzheimer's disease, Parkinson's disease, multiple sclerosis, central nervous system infections, and glioblastoma.

Example

[0147] Example The following examples are provided by way of illustration and are not intended to limit the claimed invention. In the following examples, vacuum concentration is carried out at 500 - 600 mmHg unless otherwise specified. The following abbreviations are used: DCM = dichloromethane; MeOH = methanol; EtOH = ethanol; AcOH = acetic acid; EtOAc and AcOEt = ethyl acetate; T3P® = 50% 1-propanephosphonic anhydride in EtOAc; MTBE = methyl t-butyl ether; MEK = methyl-ethyl ketone (2-butanone); MIBK = methyl-isobutyl ketone (4-methyl-2-pentanone).

[0148] Powder X-ray diffraction patterns were collected at ambient conditions (approximately 20°C) on a PANalytical X'Pert PRO X-ray diffractometer (Malvern Panalytical, Malvern, UK) using CuKα radiation (45 kV, 40 mA), a θ-θ goniometer, a collecting mirror, a divergence slit (½″), Soller slits (4 mm) in both the incident and diverging beams, a transmission type thin section sample stage (Kapton® polyimide, 12.7 μm thick film), and a PIXcel detector. The software used for data collection was X'Pert Data Collector, version 2.2f, and the data were presented using X'Pert Data Viewer, version 1.2d. The data collection range was 0.202004°s -1 The continuous operation speed was 2.994 to 35°2θ.

[0149] Differential scanning calorimeter data were collected on a PerkinElmer Pyris 6000 DSC equipped with a 45 position sample holder. Certified indium was used to verify the energy and temperature calibration of the instrument. In the following examples, a predetermined amount of sample (0.5-3.0 mg) was placed in an aluminum pan with small holes and heated at 20°C / min from 30°C to 350°C or ramped as specified by the test. A purge of 20 mL / min of dry nitrogen was maintained over the sample. Instrument control, data acquisition and data analysis were performed by Pyris Software v11.1.1 revision H.

[0150] Thermogravimetric analysis data were collected on a PerkinElmer Pyris 1 TGA equipped with an autosampler at 20 positions. The temperature of this apparatus was calibrated using certified weights as well as certified Almel and certified Parkaloy. In the following examples, a predetermined amount of sample (~5 mg) was loaded into an aluminum crucible with a pre-set tare value and heated from ambient temperature to 400 °C at 20 °C / min, unless otherwise specified. A nitrogen purge at 20 mL / min was maintained over the sample. Apparatus control, data acquisition, and data analysis were performed using Pyris Software v11.1.1 Revision H.

[0151] Example 1: Synthesis of the Compound of Formula IV The compound of Formula IV is purchased from a commercial source or prepared by the method described below or by other methods known in the art. [Chemical formula]

[0152] Ethyl 1H-pyrazole-4-carboxylate (23.5 g, 1 equiv) and acetone (587 mL) were charged into a round-bottom flask at 20 - 25 °C under a N2 atmosphere, and the mixture was stirred for 10 minutes, after which K2CO3 (70.4 g, 3 equiv) was added. The whole reaction mixture was cooled to 0 - 5 °C, and benzyl bromide (28.66 g, 1.1 equiv) was added very slowly at 0 - 5 °C over 15 minutes. The reaction mixture was warmed to 20 - 25 °C, heated to 50 - 60 °C, and maintained at that temperature for 3 hours. After the reaction was complete (monitored by HPLC), the reaction mixture was concentrated under reduced pressure at 45 - 50 °C, quenched with 10% NaOH, and extracted with DCM (117 mL). The aqueous layer was separated, back-extracted with DCM (117 mL), and the combined organic layers were dried over sodium sulfate and concentrated under reduced pressure at 45 - 50 °C. Petroleum ether or n-heptane (117 mL) was added to the concentrate, stirred for 1 hour, filtered, and concentrated under reduced pressure at 40 - 45 °C for 12 hours to yield ethyl 1-benzyl-1H-pyrazole-4-carboxylate (32.5 g).

[0153] Ethyl 1-benzyl-1H-pyrazole-4-carboxylate (30 g) and methanol (300 mL) were charged into a 3 L round-bottom flask, and the resulting solution was stirred at 24 °C for 10 minutes. Subsequently, KOH (14.6 g, 2 equivalents) was added, and the mixture was heated to 65 - 70 °C and maintained for 4 hours. After the reaction was completed (judged by HPLC), the reaction mixture was concentrated under reduced pressure to 40 - 60 mL at 45 - 50 °C. The resulting residue was dissolved in water (300 mL) and extracted with DCM (2×150 mL). The aqueous layer was separated and acidified to pH 2 with 6N HCl. The precipitated solid was filtered, washed with water (30 mL), and concentrated under reduced pressure at 45 - 50 °C for 12 hours to afford 1-benzyl-1H-pyrazole-4-carboxylic acid (19.5 g) as a light brown solid with a purity of 99.1% by HPLC. 1 1H NMR (400 MHz, DMSO-d6): δ 5.36 (s, 2H), 7.26 - 7.37 (m, 5H), 7.83 (s, 1H), 8.38 (s, 1H), 12.33 (broad s, 1H).

[0154] Example 2: Synthesis of the compound of formula III The compound of formula III was prepared as described below.

Chemical formula

[0155] DCM (285 mL) and 4-aminomethyl-benzonitrile hydrochloride (19.2 g, 1.2 equivalents) were charged into a 3 L round-bottom flask, and the mixture was cooled to 0 °C. Triethylamine (39.4 g, 3 equivalents) was added at 0 °C, and the resulting mixture was stirred for 30 minutes. Next, 1-benzyl-1H-pyrazole-4-carboxylic acid (19 g, 1 equivalent) was added at 0 - 5 °C, and the temperature was raised to 20 - 25 °C. 50% Ethyl acetate solution of 1-propanephosphonic anhydride (T3P®, Spectrochem, 72 mL, 1.28 equivalents) was added, and the mixture was stirred at 20 - 25 °C for 3 hours. After the reaction was completed, water (95 mL) was added and stirred for 10 - 15 minutes, and the organic layer was separated. The aqueous layer was extracted again with DCM (95 mL), and the combined organic layers were washed with water (95 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure to about 40 mL. Then, acetone (95 mL) was added, and the mixture was co-distilled until only 20 - 30 mL remained in the pot. Then, water (285 mL) was added, and the mixture was stirred at 20 - 25 °C for 1 hour. The resulting solid was filtered, washed with acetone:water (1:3 v / v, 10 mL), and suction filtered and dried under reduced pressure at 45 - 50 °C for 12 hours to give 1-benzyl-N-(4-cyano-benzyl)-1H-pyrazole-4-carboxamide (26.9 g, 94%) as a bright brown solid with a purity of 98.51% by HPLC. 1 1H NMR (400 MHz, DMSO-d6): δ 4.49 (d, J = 5.9 Hz, 2H), 5.37 (s, 2H), 7.27 - 7.38 (m, 5H), 7.48 (d, J = 8.1 Hz, 2H), 7.79 (d, J = 8.1 Hz, 2H), 7.95 (s, 1H), 8.31 (s, 1H), 8.77 (t, J = 5.9 Hz, 1H).

[0156] Example 3: Synthesis of the compound of formula II The compound of formula II is prepared as described below.

Chemical formula

[0157] Ethanol (250 mL) and 1-benzyl-N-(4-cyanobenzyl)-1H-pyrazole-4-carboxamide (25 g) were charged into a 1 L round-bottom flask at 20 - 25 °C under a nitrogen atmosphere. Hydroxylamine hydrochloride (16.3 g, 3 equivalents) and triethylamine (24.64 g, 3 equivalents) were added to the reaction mixture at 20 - 25 °C. The mixture was then heated to 60 - 65 °C and maintained at that temperature for 7 hours. After the reaction was complete, the mixture was concentrated under reduced pressure to about 30 - 50 mL at 45 - 50 °C. Then water (250 mL) was added and the mixture was stirred at ambient temperature for 30 minutes. The resulting solid was filtered, washed with water (125 mL), suction filtered to dryness, and further dried under reduced pressure at 45 - 50 °C for 12 hours to give 1-benzyl-N-(4-(N-hydroxycarbamimidoyl)benzyl)-1H-pyrazole-4-carboxamide (25.5 g, 92.3% yield) as a pale yellow solid with a purity of 95.41% by HPLC. 1 1H NMR (400 MHz, DMSO-d6): δ 4.40 (d, J = 5.9 Hz, 2H), 5.34 (s, 2H), 5.76 (broad s, 2H), 7.25 - 7.37 (m, 7H), 7.61 (d, J = 8.2 Hz, 2H), 7.91 (s, 1H), 8.27 (s, 1H), 8.63 (t, J = 5.9 Hz, 1H), 9.57 (broad s, 1H).

[0158] Example 4: Synthesis of the compound of formula I The compound of formula I is prepared as described below.

Chemical formula

[0159] Acetic acid (2100 g) and 1-benzyl-N-(4-(N-hydroxycarbamimidoyl)benzyl)-1H-pyrazole-4-carboxamide (200 g) were stirred in a hydrogenation apparatus at 25 °C for 10 - 15 minutes.

[0160] Raney nickel (40 g) and water (1 volume) were stirred in a flask and allowed to settle for 5 minutes. The water was decanted, another volume of water was added, stirred, allowed to settle for 5 minutes, and then decanted. Acetic acid (1 volume) was added, the mixture was stirred for 10 minutes, then allowed to settle for 5 minutes, and then decanted. The Raney nickel was charged into a hydrogenation apparatus together with acetic acid (1 volume). The reaction mixture was heated to 60 °C and hydrogen was applied for 30 minutes (10 Kg pressure). The resulting mixture was cooled to ambient temperature and the resulting solid was suction filtered over Celite® for 30 minutes. The solid was washed with MeOH (784 g), concentrated to 1 - 2 volumes, and charged with EtOAc (2 L). The mixture was stirred at 25 °C for 1 hour, suction filtered, washed with EtOAc (400 g), and suction dried for 2 hours. The product was further dried at 25 °C for 2 hours under reduced pressure (<300 mmHg), followed by drying at 45 °C for 12 hours under reduced pressure (<300 mmHg) to yield 1 - benzyl - N - (4 - carbamimidoylbenzyl) - 1H - pyrazole - 4 - carboxamide acetate (220 g) as the crude product.

[0161] Example 5: Purification The dried product (219 g) was cooled to 25 °C, charged into a round - bottom flask with water (2190 mL), and stirred for 10 minutes to form a slurry. The slurry was heated to 55 °C over 20 minutes, stirred at that temperature for 1 hour, cooled to 25 °C over 20 minutes with stirring, and further stirred at 25 °C for 30 minutes. The solid was filtered, washed with water (220 mL), and suction dried for 2 hours. The product was dried again at 45 °C for 12 hours under reduced pressure (<300 mmHg), cooled to 25 °C, and charged into a round - bottom flask. Anhydrous ethanol (1250 g) and acetic acid (183 g) were added thereto, the mixture was heated to the reflux temperature (75 °C) over 30 minutes, and reflux was maintained for 30 minutes. Then the mixture was slowly cooled to 25 °C over 30 minutes, stirred at 25 °C for 45 minutes, filtered, washed with EtOH, and suction dried at 25 °C for 2 hours. Then the product was dried at 45 °C for 10 hours under reduced pressure (<300 mmHg).

[0162] The dried product (143 g) was cooled to 25 °C and charged into a round-bottom flask together with absolute ethanol (1027 g) and acetic acid (150 g). The mixture was heated to the reflux temperature (75 °C) over 30 minutes and the reflux was maintained for 1 hour. Then the mixture was slowly cooled to 25 °C over 45 minutes, stirred at 25 °C for 45 minutes, filtered, washed with EtOH, and dried under suction at 25 °C for 2 hours. Then the product was dried at 45 °C for 12 hours under reduced pressure (<300 mmHg) to give purified 1-benzyl-N-(4-carbamimidoylbenzyl)-1H-pyrazole-4-carboxamide acetate (126 g, 56% yield, purity 99.6% by HPLC) having less than 30 ppm of Nickel.

[0163] Example 6: Pure Crystal Form 1 A mixture of crude 1-benzyl-N-(4-carbamimidoylbenzyl)-1H-pyrazole-4-carboxamide acetate (Compound 1, 10 g) in MeOH (450 mL) was charged into a 2 L round-bottom flask and the mixture was heated to 50 - 55 °C to obtain a clear solution. The solution was maintained at 50 - 55 °C for 30 minutes, filtered, and charged into the reactor at 50 - 55 °C. MTBE (450 mL) was slowly added at 50 - 55 °C and the mixture was cooled to 25 °C over 1 hour. A white suspension was observed as it cooled. MTBE (450 mL) was slowly added at 20 - 25 °C and the resulting mixture was stirred for 16 hours, filtered, and washed with MTBE (10 mL). The product was dried at 50 - 55 °C for 24 hours under reduced pressure to give anhydrous crystalline polymorphic form ( "Form 1") of pure 1-benzyl-N-(4-carbamimidoylbenzyl)-1H-pyrazole-4-carboxamide acetate as an off-white solid (8.25 g, 82.5% yield). The purity of the product was >99% by HPLC and it contained less than 14.5 ppm of nickel. 11H NMR (300 MHz, DMSO-d6): δ 1.71 (s, 3H), 4.47 (d, J = 5.4 Hz, 2H), 5.36 (s, 2H), 7.26 - 7.37 (m, 5H), 7.46 (d, J = 7.8 Hz, 2H), 7.74 (d, J = 7.8 Hz, 2H), 7.92 (s, 1H), 8.29 (s, 1H), 8.77 (broad, 1H), 10.34 (broad, 3H). 13 13C NMR (75 MHz, DMSO-d6): δ 24.7, 41.7, 55.0, 118.4, 127.4 (2C), 127.5 (2C), 127.8 (2C), 128.2 (2C), 128.6 (2C), 131.6, 136.8, 145.2, 161.8, 165.7, 176.5. The powder X-ray diffraction (XRPD) pattern included peaks at 10.0, 18.1, 18.6, 20.1, and 23.9 degrees, ±0.5 degrees, ±0.2 degrees, or ±0.1 degrees, 2θ, (CuKα radiation) and was consistent with the XRPD pattern of Figure 1. From the thermal tests, the DSC thermogram showed a single temperature profile with only a major melting endotherm at 251 °C and a single weight loss indicating the disappearance of acetate (Figure 2).

[0164] Example 7: Pure Crystalline Form 2 (A) Crystallization : Weighed 53 mg of Compound 1 into a vial, dissolved it in 3:1 MeOH:H2O (1 mL, 20 volumes) at 55 °C, and clarified it into a crystallization tube at 50 °C.

[0165] Filled the solution with MTBE (1 mL) and maintained it at 50 °C for about 10 minutes. The solution developed into a suspension within 3 minutes of MTBE addition. The mixture was cooled to 25 °C over about 1 hour and further equilibrated for 1 hour but remained a suspension. The solid was separated by filtration and vacuum dried at 50 °C for about 16 hours to produce an off-white solid with a recovery rate of 62.3%. The powder X-ray diffraction (XRPD) pattern included peaks at 4.2, 5.6, 8.4, 17.8, and 19.8 degrees, ±0.5, ±0.2, or ±0.1 degrees, 2θ, (CuKα radiation) and was consistent with the XRPD pattern of Figure 2.

[0166] (B) Seeding Crystallization: Compound 1 (1.008 g) was weighed into a container and filled with 3:1 MeOH:H2O (22 mL, 22 volumes). The mixture was heated to 55 °C to achieve dissolution. The solution was clarified at 50 °C and filled into a container. Seed crystals of Form II (about 10 mg) were added to this solution and observed to remain.

[0167] The container was filled with MTBE (22 mL, 22 volumes) and the mixture was cooled to 25 °C over about 1 hour. As it cooled, it was observed to grow into a fine off-white suspension. The mixture was further equilibrated for 45 minutes and then filled with additional MTBE (22 mL, 22 volumes). The suspension was equilibrated at 25 °C for a further 16.5 hours. The solid was separated by filtration and vacuum dried at 50 °C for about 23.5 hours to give a white, free-flowing powder solid in 73.5% yield. The powder X-ray diffraction (XRPD) pattern contained peaks at 4.2, 5.6, 8.4, 17.8, and 19.8 degrees, ±0.5, ±0.2, or ±0.1 degrees, 2θ, (CuKα radiation) and was consistent with the XRPD pattern of Figure 2. From the thermal tests (Figure 9), a DSC thermograph typical of Form II with a prominent endotherm at 158 °C was revealed prior to the major melting endotherm at 254 °C. The TGA thermograph showed a weight loss of 4.067 wt% from 40 - 105 °C, suggesting that there was almost stoichiometric loss of water prior to the weight loss due to the disappearance of acetate.

[0168] Example 8: Pure Crystalline Form 3 (A) Via CH3CN: Compound 1 (25 mg) was weighed into a crystallization tube and filled with acetonitrile (MeCN) in 0.1 mL (4 volume) aliquots up to 0.5 mL (20 volumes) while noting the immediate observations. The mixture was stirred with a magnetic stirrer bar, heated to 50 °C, and observations were noted after 4 hours. Since it did not dissolve clearly, the volume of the solvent was doubled to 1 mL (40 volumes) and immediate observations were noted. The mixture was cooled to 25 °C and equilibrated for 65 hours. Observations were noted and half of the mixture was separated. Before separating the remainder, the mixture was heated to 50 °C and equilibrated for 6 hours.

[0169] Before analysis by XRPD test, the solid was vacuum dried at 50 °C for 17 hours. The powder X-ray diffraction (XRPD) pattern included peaks at 7.7, 10.7, 20.1, 23.7, and 24.3 degrees and was consistent with the XRPD pattern in Figure 3. From the thermal test of Form III (Figure 10), there was evidence of a small shoulder at 234 °C and a single endothermic peak at 249 °C in the DSC thermograph. There was a weight loss due to disproportionation of acetate during melting by TGA.

[0170] (B) Via MeOH: Compound 1 (1.050 g) was weighed into a flask and filled with MeOH (50 mL, 50 volumes). The mixture was heated to 50 °C and maintained for 20 minutes. Since it only partially dissolved, it was further heated to 60 °C and additional MeOH (10 mL, total 60 volumes) was added to achieve dissolution. The solution was clarified and reduced to dryness in vacuo and further vacuum dried at 50 °C for 16.5 hours to yield a pink, large quantity of low-density solid with a recovery rate of 76.1%.

[0171] Example 9: Pure Crystal Form 4 (A) From Form I: Compound 1 (211 mg) was weighed into a vial and filled with MeOH (8.44 mL, 40 volumes). The mixture was heated to 50 °C and maintained for 1 hour. Since it only partially dissolved, it was further heated to achieve dissolution. The solution was clarified and filled into crystallization tubes at 50 °C in 1 mL aliquots. EtOAc was filled to 1 mL in 0.1 mL aliquots. The mixture was cooled to 20 °C over 2 hours and further equilibrated at 20 °C for 19.25 hours.

[0172] The solid was vacuum dried at 50 °C for about 22.5 hours. The powder X-ray diffraction (XRPD) pattern included peaks at 18.5, 19.6, and 23.7 degrees 2θ, ±0.5, ±0.2, or ±0.1 degrees and was consistent with the XRPD pattern in Figure 4. From the thermal test of Form IV, a single temperature profile with only a major melting endotherm at 253 °C in the DSC thermograph (Figure 11) and a small weight loss of about 0.40 wt% from 100 - 150 °C before disappearance of acetate were revealed.

[0173] (B) From amorphous: Amorphous compound 1 (4 × 50 mg) and Form 4 (4 × 2 mg) were weighed into four crystallization tubes, and four types of solvents were charged (THF, EtOAc, MIBK, and MEK: 2 mL, 40 vol). Before cooling the mixture to 25 °C, it was stirred with a magnetic stirrer bar at 50 °C for about 21 h. Before heating the suspension to 50 °C, it was equilibrated at 25 °C for about 7.5 h. Before cooling the suspension to 25 °C, it was equilibrated at 50 °C for about 16 h, and the solid was separated by filtration.

[0174] The solid was vacuum dried at 50 °C for about 72 h. XRPD demonstrated that the solid recovered from THF or EtOAc was Form 1, while that recovered from MIBK or MEK was Form 4.

[0175] Example 10: Pure amorphous Form 5 Compound 1 (495 mg) was weighed into a flask containing deionized water (250 mL), and dissolution was achieved by stirring at 45 °C. The solution was clarified in a 3 L flask, frozen, and lyophilized for about 9 h. A white and large amount of solid was separated with a recovery rate of about 83.6%. From the test by XRPD, it was found that the solid was mainly amorphous, but a trace amount of Form II was present.

[0176] The above process was repeated, and the solid was lyophilized for about 6.25 h, producing a white and large amount of solid with a recovery rate of 70.5%. From the test by XRPD, it was found that the substance was mainly amorphous (Figure 5). From the thermal test (Figure 12), a small endothermic event at about 112 °C similar to Form II, and the onset and peak of the main endotherms at about 229 °C and 240 °C, which were slightly decreased compared to Form II, became apparent. A weight loss of 1.300 wt% was observed from 40 to 80 °C, which was mainly the remaining surface moisture. A second significant weight loss of 15.837 wt% from 170 to 240 °C corresponding to melting indicated the disappearance of acetate (15.01 wt%), and perhaps the disappearance of further trapped water or a small decomposition event led to the onset of decomposition from 250 °C.

[0177] Example 11: Pressure and Grinding Stability In the manufacturing process of solid dosage forms, high compression forces and / or grinding may be required, which may cause changes in the crystal structure of some compounds. Forms I and II of Compound 1 were compressed at a force of 11×10 5 N for about 25 - 28 hours to evaluate the effect of compression force on the form stability of these crystal forms.

[0178] From the XRPD tests, it was found that the diffraction resolution of Form I decreased slightly. From the tests of Form II, as shown in Figure 6, it was found that the diffraction pattern changed significantly to a pattern with reduced crystallinity. From the thermal tests of Form I, it was found that there was no change in the TGA thermogram after pressurization. In the DSC thermogram after pressurization, the intensity and temperature of the endothermic melting decreased. From the thermal tests of Form II, it was found that the moisture content increased slightly according to the TGA thermogram, but this may have been due to the deterioration of the sample after separation rather than the pressurization effect. From the DSC thermogram, a decrease in intensity was revealed after pressurization, but an increase in the main endothermic melting temperature and an additional significant endotherm at about 150 °C were revealed.

[0179] Samples of Form I and Form II (about 100 mg each) were subjected to a grinding operation to evaluate the effect on the solid form and the observed microscopy. Attention was paid to whether an amorphous component was generated, and in particular, with respect to the hydrate of Form II, there was interest in examining the effect on the moisture content of the batch.

[0180] It was clear from the dataset of both samples after grinding that there were no changes either thermally or crystallographically. Form II showed small changes in reflection resolution and intensity, as shown in Figure 7, but neither sample showed an amorphous component either thermally or by XRPD compared to the input. The most significant change regarding the pressurized material was that Form II had an increase in moisture content to 3.93 wt% (from 2.76 wt%) at the time of manufacture, which was consistent with the expected hydrate.

[0181] A microscopic test was performed comparing both samples to their initial state. After milling, both samples were very well dispersed in silicone oil. Form II had more particles maintaining a particle size of 5 - 10 μm, although most of the solids were <5 μm. Form I was broken down into solids of mainly <5 μm particles (mostly 1 - 2 μm) that were very fine and evenly dispersed, as would be expected assuming a relatively small particle size was observed in the input lot. In summary, under manual stress, Form I showed better stability than the hydrated Form II.

Claims

1. A crystalline form of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate, substantially free of other polymorphic forms of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate.

2. The crystalline form is anhydrous 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate, characterized by a melting point of about 253 °C and a water solubility of about 8.3 mg / mL at 25 °C (“Form I”), the crystalline form according to claim 1.

3. Form I is characterized by a powder X-ray diffraction (XRPD) pattern comprising peaks at 10.0, 18.1, 18.6, 20.1, and 23.9 degrees ± 0.5 degrees, 2θ, said XRPD being made using CuKα radiation, the crystalline form according to claim 2.

4. The XRPD pattern further comprises a peak at 20.5 degrees ± 0.5 degrees, 2θ, the crystalline form according to claim 3.

5. The XRPD pattern is substantially similar to the pattern of FIG. 1, the crystalline form according to claim 3 or 4.

6. The 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate is hydrated, characterized by a melting point of about 253 °C and a water solubility of about 5.5 mg / mL at 25 °C (“Form II”), the crystalline form according to claim 1.

7. Form II is characterized by an XRPD pattern comprising peaks at 4.2, 5.6, 8.4, 17.8, and 19.8 degrees ± 0.5 degrees, 2θ, said XRPD being made using CuKα radiation, the crystalline form according to claim 6.

8. The XRPD pattern further comprises a peak at 12.7 degrees ± 0.5 degrees, 2θ, the crystalline form according to claim 7.

9. The XRPD pattern is substantially similar to the pattern of FIG. 2, the crystalline form according to claim 7 or 8.

10. The 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate is an anhydride, characterized by a melting point of about 249 °C and a water solubility of about 7.2 mg / mL at 25 °C (“Form III”), the crystalline form according to claim 1.

11. Form III is characterized by an XRPD pattern comprising peaks at 7.7, 10.7, 20.1, 23.7 and 24.3 degrees ± 0.5 degrees, 2θ, said XRPD being made using CuKα radiation, the crystalline form according to claim 10.

12. The XRPD pattern further comprises peaks at 18.5 and 26.5 degrees ± 0.5 degrees, 2θ, the crystalline form according to claim 11.

13. The XRPD pattern is substantially similar to the pattern of FIG. 3, the crystalline form according to claim 11 or 12.

14. The form is characterized by having a melting point of about 251 ° C. and is characterized by an XRPD pattern comprising peaks at 18.5, 19.6 and 23.7 degrees ± 0.5 degrees, 2θ, 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate, said XRPD being made using CuKα radiation ("Form IV"), the crystalline form according to claim 1.

15. The XRPD pattern further comprises a peak at 24.6 degrees ± 0.5 degrees, 2θ, the crystalline form according to claim 14.

16. The XRPD pattern is substantially similar to the pattern of FIG. 4, the crystalline form according to claim 14 or 15.

17. Characterized by a melting point of about 240 ° C. and a water solubility of about 7.1 mg / mL in water at 25 ° C. ("Form V"), an amorphous form of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate.

18. A pharmaceutical composition for treating a plasma kallikrein-dependent disease or condition, comprising a therapeutically effective amount of the solid form according to any one of claims 1 to 17 and a pharmaceutically acceptable carrier.

19. The solid form is the form I according to any one of claims 2 to 5 and a pharmaceutically acceptable carrier, the pharmaceutical composition according to claim 18.

20. The solid form is the form II according to any one of claims 6 to 9 and a pharmaceutically acceptable carrier, the pharmaceutical composition according to claim 18.

21. The solid form is the form III according to any one of claims 10 to 13 and a pharmaceutically acceptable carrier, the pharmaceutical composition according to claim 18.

22. The pharmaceutical composition according to claim 18, wherein the solid form is Form IV according to any one of claims 14 to 16 and a pharmaceutically acceptable carrier.

23. The pharmaceutical composition according to claim 18, wherein the solid form is Form V according to claim 17 and a pharmaceutically acceptable carrier.

24. The method for producing Form I according to any one of claims 2 to 5, comprising contacting the solid form of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate with a solvent selected from the group consisting of methanol, ethanol, isopropanol, tetrahydrofuran, 2-methyl-tetrahydrofuran, acetone anhydrous, dichloromethane, diethyl ether, 3-methyl-1-butanol, and nitromethane and mixtures thereof for a time sufficient to form Form I.

25. The method according to claim 24, wherein the solvent is methanol.

26. The method according to claim 24 or claim 25, wherein the solvent further comprises a solvent selected from the group consisting of tetrahydrofuran, t-butyl-methyl ether, and heptane.

27. The method for producing Form II according to any one of claims 6 to 9, comprising contacting the solid form of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate with a solvent selected from the group consisting of water and a solvent selected from the group consisting of isopropanol, tetrahydrofuran, acetone, and ethyl acetate, or a mixture thereof, for a time sufficient to form Form II.

28. The method according to claim 27, wherein the mixture comprises at least about 10% water.

29. The method according to claim 27 or claim 28, wherein the solvent is ethyl acetate.

30. (a)Dissolving the solid form of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate in a sufficient amount of water to form a solution; (b)Freezing the solution; and (c)Lyophilizing the product of step (b), which is the method for producing Form V according to claim 17.

31. A process for producing Form III according to any one of claims 10 to 13, comprising contacting the solid form of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate with acetonitrile for a time sufficient to produce Form III.

32. The crystalline form according to claim 1, having less than 10% by weight of any other polymorphic form.

33. The crystalline form according to claim 32, having less than 5% by weight of any other polymorphic form.

34. The crystalline form according to claim 32 or claim 33, having less than 1% by weight of any other polymorphic form.

35. A method for treating a plasma kallikrein-dependent disease or condition, said method comprising administering an effective amount of the crystalline form according to any one of claims 1 to 17, or the pharmaceutical composition according to any one of claims 18 to 23, to a subject in need thereof. Treatment method.

36. The plasma kallikrein-dependent disease or condition is diabetic macular edema, diabetic retinopathy, hereditary angioedema due to C1 inhibitor deficiency, acute liver injury, inflammation, anaphylaxis, chemically sensitized kidney injury, ischemic stroke, hemorrhagic stroke, hypertension, vascular complications of hypertension, retinopathy, nephropathy, cerebral vascular edema, pulmonary hypertension, inflammation, pain, acute myocardial infarction, deep vein thrombosis, complications resulting from fibrinolytic therapy, angina pectoris, angioedema, sepsis, arthritis, complications of cardiopulmonary bypass, capillary leak syndrome, inflammatory bowel disease, diabetes, diabetic retinopathy, diabetic macular edema, diabetic nephropathy, diabetic neuropathy, age-related macular degeneration, retinal vein occlusion, cerebral edema, ischemia-reperfusion injury, cancer-related angiogenesis, asthma, anaphylaxis, and Alzheimer's disease, Parkinson's disease, multiple sclerosis, central nervous system infections, and cerebral vascular complications of glioblastoma, selected from the group consisting of, the method according to claim 35.

37. Use of a crystalline form of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate substantially free of other polymorphic forms thereof for the treatment of plasma kallikrein-dependent diseases or conditions.

38. The use according to claim 37, wherein the solid form is the form according to any one of claims 1 to 17.

39. Use of a solid form of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate, substantially free of other polymorphic forms of 1-benzyl-N-(4-carbamimidoyl-benzyl)-1H-pyrazole-4-carboxamide acetate, for the manufacture of a medicament for the treatment of a plasma kallikrein-dependent disease or condition.

40. Use according to claim 39, wherein the solid form is a form according to any one of claims 1 to 17.

41. Crystalline form according to any one of claims 3, 4, 7, 8, 11, 12, 14, 15, wherein the XRPD peaks are within ±0.2 degrees 2θ of the stated peak positions.

42. Crystalline form according to claim 41, wherein the XRPD peaks are within ±0.1 degrees 2θ of the stated peak positions.

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