WO2021211938A1 - Solid forms of inhibitors of plasma kallikrein - Google Patents

Abstract

The present invention provides compounds and compositions thereof which are useful as inhibitors of Plasma Kallikrein (pKal) and which exhibit desirable characteristics for the same.

Description

SOLID FORMS OF INHIBITORS OF PLASMA KALLIKREIN

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and the benefit of U.S. Provisional Patent

Application Serial No. 63/011,795, filed on April 17, 2020, entitled “SOLID FORMS OF INHIBITORS OF PLASMA KALLIKREIN,” the disclosure of which is incorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION

[0002] Plasma kallikrein (pKal) is a serine protease zymogen in blood that is converted to its catalytically active form by coagulation factor Xlla, and contributes to the innate inflammatory response and intrinsic cascade of blood coagulation. The mechanisms that lead to the activation of this pathway in vivo include interactions with polyphosphates released from activated platelets and deficiency of Cl inhibitor (Cl-INH), the primary physiological inhibitor of pKal. pKal-mediated cleavage of high-molecular weight kininogen generates the potent vasodilator and pro-inflammatory nonapeptide bradykinin (BK), which activates the bradykinin 2 receptor. Subsequent cleavage of BK by carboxypeptidases generates des-Arg9-BK, which activates the B1 receptor. Both B1 and B2 receptors are expressed by vascular, glial, and neuronal cell types, with the highest levels of retinal expression detected in the ganglion cell layer and inner and outer nuclear layers. Activation of B1 and B2 receptors causes vasodilation and increases vascular permeability.

[0003] pKal is also associated with a number of disorders, such as hereditary angioedema (HAE), an autosomal dominant disease characterized by painful, unpredictable, recurrent attacks of inflammation affecting the hands, feet, face, abdomen, urogenital tract, and the larynx. Prevalence for HAE is uncertain but is estimated to be approximately 1 case per 50,000 persons without known differences among ethnic groups. HAE is caused by deficient (Type I) or dysfunctional (Type II) levels of Cl-INH, which inhibits pKal, bradykinin, and other serine proteases in the blood. Individuals with hereditary angioedema (HAE) are deficient in Cl-INH and consequently undergo excessive bradykinin generation, which in turn cause painful, debilitating, and potentially fatal swelling attacks. If left untreated, HAE can result in a mortality rate as high as 40% primarily due to upper airway obstruction. Consequently, there is a great need in the art for effective inhibitors of pKal. [0004] Chemical compounds can form one or more different pharmaceutically acceptable salts and/or solid forms, including amorphous and polymorphic crystal forms. Individual salts and solid forms of bioactive chemical compounds can have different properties. There is a need for the identification and selection of appropriate salts and/or solid forms of bioactive chemical compounds (including appropriate crystalline forms, where applicable) for the development of pharmaceutically acceptable dosage forms for the treatment of various diseases or conditions associated with pKal.

SUMMARY OF THE INVENTION

[0005] The present disclosure provides novel salts and solid forms useful as inhibitors to plasma kallikrein (pKal). In general, salt forms or free base forms, and pharmaceutically acceptable compositions thereof, are useful for treating or lessening the severity of a variety of diseases or disorders as described in detail herein.

BRIEF DESCRIPTION OF THE DRAWINGS

[0006] Figure 1 provides X-ray powder diffraction (XRPD) pattern of Compound 6

Pattern 1 (oxalate).

[0007] Figure 2 provides TGA/DSC curves for Compound 6 Pattern 1 (oxalate).

[0008] Figure 3 provides X-ray powder diffraction (XRPD) pattern of Compound 6

Pattern 4 (oxalate).

[0009] Figure 4 provides TGA/DSC curves for Compound 6 Pattern 4 (oxalate).

DETAILED DESCRIPTION OF THE INVENTION

General Description of Certain Aspects of the Invention:

[0010] PCT patent application publication number WO2019/178129, filed March 12,

2019 and published September 19, 2019 (“the ‘129 publication”), the entirety of which is hereby incorporated herein by reference, describes certain plasma kallikerin (pKal) inhibitor compounds. Such compounds include N-((7-chloro-8-fluoroimidazo[l,5-a]pyridin-l- yl)methyl)-l-((6-cyclopropylimidazo[l,2-a]pyridin-2-yl)methyl)-lH-l, 2, 3-tri azole-4- carboxamide:

[0011] Compound 1, which is a free base, is one of many compounds identified as a small molecule inhibitor of pKal in the ‘129 publication. In the ‘129 publication, Compound 1 is identified as compound 1-148 and its synthesis is described in detail at Example 148, which is reproduced herein for ease of reference.

[0012] Compound 1 has shown potency against plasma kallikrein in an in vitro assay

(See, e.g., Table 1 of the ‘129 publication). For example, the ‘129 publication reports that Compound 1 has an EC 50 < 1 nM as measured in an in vitro kallikrein kinase assay. Accordingly, Compound 1 is useful for treating one or more disorders associated with activity of pKal.

[0013] The present disclosure refers to various free base solid forms of Compound 1, salt forms of Compound 1 and solid forms thereof, pharmaceutical compositions thereof, and methods of preparing solid forms of Compound 1 and salts and solid forms thereof. Salt forms and solid forms (e.g., crystalline solid forms) impart or may impart characteristics such as improved aqueous solubility, stability, absorption, bioavailability, and ease of formulation. As used herein, unless otherwise indicated the term “salt” refers to a salt or co-crystal of two or more (e.g., two) component molecules (e.g., Compound 1 and a co-former). In the combination of an acid and a base compound for the preparation of a solid form, a A pK_a (pKa(base)- pK_a(acid)) > 1 generally will permit the formation of a salt compound where the two compounds are ionized. Where this threshold is not met, non-ionic interactions (e.g., hydrogen bonds) can still occur between neutral acid and the base compounds to form, e.g., a co-crystal. In some embodiments, a provided solid form is a salt. In other embodiments, a provided solid form is a co-crystal.

Compound 3 (L-malic Acid x Compound 1 )

[0014] According to one embodiment, the present disclosure refers to a chemical species Compound 2 comprising Compound 1 and L-malic acid:

Compound 3

Compound 4 (Succinic acid x Compound 1)

[0015] According to one embodiment, the present disclosure refers to a chemical species Compound 4 comprising Compound 1 and succinic acid:

Compound 5 (Fumaric acid x Compound 1)

[0016] According to one embodiment, the present disclosure refers to a chemical species Compound 5 comprising Compound 1 and fumaric acid:

Compound 5

Compound 6 (Oxalic acid x Compound 1)

[0017] According to one embodiment, the present disclosure provides a chemical species (e.g., a solid form) Compound 6 comprising Compound 1 and oxalic acid:

Compound 6 [0018] In one embodiment, a solid form of Compound 6 has a stoichiometry of

(Compound 1): (oxalic acid) that is about 1:1. As used herein, the term “about”, when used in reference to a stoichiometric ratio refers to 1:(1±0.2) ratio of (Compound 1): (co-former, e.g., an acid), e.g., a 1:(1±0.2) ratio, a 1:(1±0.1) ratio, or a 1:(1±0.05) ratio.

[0019] In some embodiments, the present disclosure provides Compound 6 substantially free of impurities. As used herein, the term “substantially free of impurities” means that the compound contains no significant amount of extraneous matter. Such extraneous matter may include excess oxalic acid, excess Compound 1, residual solvents, or any other impurities that may result from the preparation of, and/or isolation of, Compound 6. In certain embodiments, at least about 95% by weight of Compound 6 is present. In still other embodiments of the invention, at least about 99% by weight of Compound 6 is present.

[0020] According to one embodiment, Compound 6 is present in an amount of at least about 97.0, 97.5, 98.0, 98.5, 99.0, 99.5, or 99.8 weight percent where the percentages are based on the total weight of the composition. According to another embodiment, Compound 6 contains no more than about 3.0 area percent HPLC of total organic impurities and, in certain embodiments, no more than about 1.5 area percent HPLC total organic impurities relative to the total area of the HPLC chromatogram. In other embodiments, Compound 6 contains no more than about 1.0 area percent HPLC of any single impurity; no more than about 0.6 area percent HPLC of any single impurity, and, in certain embodiments, no more than about 0.5 area percent HPLC of any single impurity, relative to the total area of the HPLC chromatogram.

[0021] The structure depicted for Compound 6 is also meant to include all tautomeric forms of Compound 6. Additionally, structures depicted here are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structure except for the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a ¹³C- or ¹⁴C-enriched carbon are within the scope of this invention.

[0022] It has been found that Compound 6 can exist in a variety of solid forms.

Exemplary such forms include polymorphs such as those described herein.

[0023] In some embodiments, Compound 6 is amorphous. In some embodiments,

Compound 4 is amorphous, and is substantially free of crystalline Compound 6. [0024] In certain embodiments, Compound 6 is a crystalline solid. In other embodiments, Compound 6 is a crystalline solid substantially free of amorphous Compound 6. As used herein, the term “substantially free of amorphous Compound 6” means that the compound contains no significant amount of amorphous Compound 6. In certain embodiments, at least about 95% by weight of crystalline Compound 6 is present. In still other embodiments of the invention, at least about 99% by weight of crystalline Compound 6 is present.

[0025] It has been found that Compound 6 can exist in at least two polymorphic forms. In some embodiments, the present invention provides a polymorphic form of Compound 6 referred to herein as Pattern 1. In some embodiments, the present invention provides a polymorphic form of Compound 6 referred to herein as Pattern 4.

Compound 6 Pattern 1

[0026] In some embodiments, Compound 6 Pattern 1 has at least 1, 2, 3, 4 or 5 X-ray

Powder Diffraction (XRPD) peaks selected from the angles (2 theta ± 0.2) listed in Table 1 below.

Table 1. XRPD Peak Positions for Compound 6 Pattern 1

[0027] In some embodiments, Compound 6 Pattern 1 is characterized by an X-ray powder diffraction (XRPD) pattern having diffractions at angles (2 theta ± 0.2) and corresponding d-spacing (angstroms ± 0.2) of:

Table 2. XRPD Peak Positions and d-Spacing for Compound 6 Pattern 1

[0028] In some embodiments, Compound 6 Pattern 1 is characterized in that it has one or more peaks in its X-ray powder diffraction pattern selected from those at about 4.7,

6.1, 18.7, 24.0, 24.2, and 24.6. In some embodiments, Compound 6 Pattern 1 is characterized in that it has two or more peaks in its X-ray powder diffraction pattern selected from those at about 4.7, 6.1, 18.7, 24.0, 24.2, and 24.6. In some embodiments, Compound 6 Pattern 1 is characterized in that it has three or more peaks in its X-ray powder diffraction pattern selected from those at about 4.7, 6.1, 18.7, 24.0, 24.2, and 24.6. In some embodiments, Compound 6 Pattern 1 is characterized in that it has four or more peaks in its X-ray powder diffraction pattern selected from those at about 4.7, 6.1, 18.7, 24.0, 24.2, and 24.6. In some embodiments, Compound 6 Pattern 1 is characterized in that it has five or more peaks in its X-ray powder diffraction pattern selected from those at about 4.7, 6.1, 18.7, 24.0, 24.2, and

24.6. In some embodiments, Compound 6 Pattern 1 is characterized in that it has all six peaks in its X-ray powder diffraction pattern selected from those at about 4.7, 6.1, 18.7, 24.0, 24.2, and 24.6. In some embodiments, Compound 6 Pattern 1 is characterized in that it has all six peaks in its X-ray powder diffraction pattern selected from those at about 4.7, 6.1,

18.7, 24.0, 24.2, and 24.6, corresponding to d-spacing (angstroms ± 0.2) of 18.88, 14.36,

4.73, 3.71, 3.67, and 3.62 (respectively).

[0029] In certain embodiments, the X-ray powder diffraction pattern of Compound 6

Pattern 1 is substantially similar to the XRPD provided in Figure 1.

[0030] Methods for preparing Compound 6 Pattern 1 are described infra. Compound 6 Pattern 4

[0031] In some embodiments, Compound 6 Pattern 4 has at least 1, 2, 3, 4 or 5 X-ray

Powder Diffraction (XRPD) peaks selected from the angles (2 theta ± 0.2) listed in Table 3 below.

Table 3. XRPD Peak Positions for Compound 6 Pattern 4

[0032] In some embodiments, Compound 6 Pattern 4 is characterized by an X-ray powder diffraction (XRPD) pattern having diffractions at angles (2 theta ± 0.2) and corresponding d-spacing (angstroms ± 0.2) of:

Table 4. XRPD Peak Positions and d-Spacing for Compound 6 Pattern 4

[0033] In some embodiments, Compound 6 Pattern 4 is characterized in that it has one or more peaks in its X-ray powder diffraction pattern selected from those at about 6.4, 11.9, 19.2, 26.1, 26.6, and 27.2. In some embodiments, Compound 6 Pattern 4 is characterized in that it has two or more peaks in its X-ray powder diffraction pattern selected from those at about 6.4, 11.9, 19.2, 26.1, 26.6, and 27.2. In some embodiments, Compound 6 Pattern 4 is characterized in that it has three or more peaks in its X-ray powder diffraction pattern selected from those at about 6.4, 11.9, 19.2, 26.1, 26.6, and 27.2. In some embodiments, Compound 6 Pattern 4 is characterized in that it has four or more peaks in its X-ray powder diffraction pattern selected from those at about 6.4, 11.9, 19.2, 26.1, 26.6, and

27.2. In some embodiments, Compound 6 Pattern 4 is characterized in that it has five or more peaks in its X-ray powder diffraction pattern selected from those at about 6.4, 11.9,

19.2, 26.1, 26.6, and 27.2. In some embodiments, Compound 6 Pattern 4 is characterized in that it has all six peaks in its X-ray powder diffraction pattern selected from those at about 6.4, 11.9, 19.2, 26.1, 26.6, and 27.2. In some embodiments, Compound 6 Pattern 4 is characterized in that it has all six peaks in its X-ray powder diffraction pattern selected from those at about 6.4, 11.9, 19.2, 26.1, 26.6, and 27.2, corresponding to d-spacing (angstroms ± 0.2) of 13.89, 7.43, 4.63, 3.42, 3.35, and 3.27 (respectively).

[0034] In certain embodiments, the X-ray powder diffraction pattern is substantially similar to the XRPD provided in Figure 3.

[0035] Methods for preparing Compound 6 Pattern 4 are described infra.

General Methods of Providing the Compounds

[0036] Compound 1 is prepared according to the methods described in detail in the

‘129 publication, the entirety of which is hereby incorporated herein by reference.

[0037] The acid addition compounds of general formula A, which formula encompasses, inter alia, Compounds 3-6, and/or particular forms thereof, are prepared from Compound 1, according to the general Scheme below.

Compound 1 Compound A

[0038] In this scheme, “Acid” represents, e.g., any of the co-formers described herein.

For instance, each of Compounds 3-6, and forms thereof, are prepared from Compound 1 by combining Compound 1 with an appropriate acid to form the product Compound. Thus, another aspect of the present disclosure refers to a method for preparing Compounds 3-6, and forms thereof, by combining Compound 1 with an appropriate acid to form the product Compound.

[0039] As described generally above, in some embodiments, the present invention provides a method for preparing Compound A:

Compound A comprising steps of: combining Compound 1:

Compound 1 with a suitable co-former (e.g., a suitable acid) and optionally a suitable solvent under conditions suitable for forming Compound A.

[0040] In some embodiments, Compound 1 is treated with a co-former selected from: oxalic acid, L-malic acid, fumaric acid, and succinic acid.

[0041] In some embodiments, a suitable co-former is oxalic acid.

[0042] A suitable solvent may be any solvent system (e.g., one solvent or a mixture of solvents) in which Compound 1 and/or an acid are soluble, or are at least partially soluble. [0043] Examples of suitable solvents useful in the present invention include, but are not limited to protic solvents, aprotic solvents, polar aprotic solvent, or mixtures thereof. In certain embodiments, suitable solvents include an ether, an ester, an alcohol, a ketone, or a mixture thereof. In some embodiments, a solvent is one or more organic alcohols. In some embodiments, a solvent is chlorinated. In some embodiments, a solvent is an aromatic solvent.

[0044] In certain embodiments, a suitable solvent is methanol, ethanol, isopropanol, t- butanol, acetonitrile, tetrahydrofuran (THF), or acetone wherein said solvent is anhydrous or in combination with water or dichloromethane (DCM). In some embodiments, suitable solvents include n-heptane, ethyl acetate, methyl ethyl ketone (MEK), tert-butyl methyl ether (TBME), isopropyl acetate (IP AC), methyl isobutyl ketone (MIBK), dimethylformamide (DMF), dimethylacetamide (DMAC), dimethylsulfoxide (DMSO), toluene, trifluorotoluene, anisole, chlorobenzene, cumene, or N-methylpyrrolidone (NMP). In some embodiments, a suitable solvent is acetone. In some embodiments, a suitable solvent is methanol. In some embodiments, a suitable solvent is ethyl acetate. In some embodiments, a suitable solvent is a combination of said solvents.

[0045] In some embodiments, the present invention provides a method for preparing a free base form of Compound 1 or Compound A, comprising one or more steps of removing a solvent and adding a solvent. In some embodiments, an added solvent is the same as the solvent removed. In some embodiments, an added solvent is different from a solvent removed. Means of solvent removal are known in the synthetic and chemical arts and include, but are not limited to, any of those described herein and in the Exemplification.

[0046] In some embodiments, a method for preparing a free base form of Compound

1 or Compound A comprises one or more steps of heating or cooling a preparation.

[0047] In some embodiments, a method for preparing a free base form of Compound

1 or Compound A comprises one or more steps of agitating or stirring a preparation.

[0048] In some embodiments, a method for preparing a free base form of Compound

1 or Compound A comprises a step of adding a suitable co-former to a solution or slurry of compound 1.

[0049] In some embodiments, a method for preparing a free base form of Compound

1 or Compound A comprises a step of adding a suitable acid to a solution or slurry of compound 1. [0050] In some embodiments, a method for preparing a free base form of Compound

1 or Compound A comprises a step of heating.

[0051] In certain embodiments, a free base form of Compound 1 or Compound A precipitates from the mixture. In another embodiment, a free base form of Compound 1 or Compound A crystallizes from the mixture. In other embodiments, a free base form of Compound 1 or Compound A crystallizes from solution following seeding of the solution (i.e., adding crystals of a free base form of Compound 1 or Compound A to the solution).

[0052] A free base form of Compound 1 or Compound A can precipitate out of the reaction mixture, or be generated by removal of part or all of the solvent through methods such as evaporation, distillation, filtration (e.g., nanofiltration, ultrafiltration), reverse osmosis, absorption and reaction, by adding a suitable anti-solvent, for example but not limited to, heptane, cumene, toluene, and TBME, by cooling or by different combinations of these methods.

[0053] As described generally above, a free base form of Compound 1 or Compound

A is optionally isolated. It will be appreciated that a free base form of Compound 1 or Compound A may be isolated by any suitable physical means known to one of ordinary skill in the art. In certain embodiments, precipitated solid free base form of Compound 1 or Compound A is separated from the supernatant by filtration. In other embodiments, precipitated solid free base form of Compound 1 or Compound A is separated from the supernatant by decanting the supernatant.

[0054] In certain embodiments, a free base form of Compound 1 or Compound A is separated from the supernatant by filtration.

[0055] In certain embodiments, an isolated free base form of Compound 1 or

Compound A is dried in air. In other embodiments isolated free base form of Compound 1 or Compound A is dried under reduced pressure, optionally at elevated temperature.

Methods of Use

[0056] In certain embodiments, compounds of the present invention (e.g., Compound

6) are for use in medicine. In some embodiments, compounds of the present invention are useful as serine protease zymogen inhibitor. In certain embodiments, compounds of the present invention are selective inhibitors of plasma kallikrein (pKal). In some embodiments, the present invention provides methods of decreasing pKal activity. Such methods include contacting pKal with an effective amount of a provided compound. Therefore, the present invention further provides methods of inhibiting pKal activity by contacting pKal with a compound of the present invention.

[0057] In some embodiments, provided compounds are useful for the treatment of diseases and disorders that may be alleviated by inhibiting (i.e., decreasing) pKal activity. By “diseases” is meant diseases or disease symptoms. Thus, the present invention provides methods of treating pKal-mediated disorders in a subject in need thereof. Such methods include administering to the subject a therapeutically effective amount of a provided compound.

[0058] Exemplary pKal-mediated disorders include edema, which refers to swelling in the whole body of a subject or a part thereof due to inflammation or injury when small blood vessels become leaky and releases fluid into nearby tissues. In some examples, the edema is hereditary angioedema (HAE). In other examples, the edema occurs in eyes, e.g., diabetic macular edema (DME). The present disclosure provides methods of inhibiting the activity of pKal. In certain embodiments, the application provides a method of inhibiting the activity of pKal in vitro via contacting any of the compounds described herein with pKal molecules in a sample, such as a biological sample. In certain embodiments, the application provides a method of inhibiting the activity of pKal in vivo via delivering an effective amount of any of the compounds described herein to a subject in need of the treatment through a suitable route.

[0059] In certain embodiments, provided methods comprise administering to a subject in need thereof (e.g., a subject such as a human patient with edema) any of the compounds described herein. In certain embodiments, the methods comprise administering Compound 6, or a pharmaceutically acceptable composition thereof, to a subject in need thereof. In some embodiments, the method comprises administering a pharmaceutical composition comprising Compound 6.

[0060] In certain embodiments, the subject to be treated by any of the methods described herein is a human patient having, suspected of having, or at risk for edema, for example, HAE or DME. A subject having an edema can be identified by routine medical examination, e.g., laboratory tests. A subject suspected of having an edema might show one or more symptoms of the disease/disorder. A subject at risk for edema can be a subject having one or more of the risk factors associated with the disease, for example, deficiency in Cl inhibitor (Cl-INH) as for HAE.

[0061] In certain embodiments, provided herein are methods of alleviating one or more symptoms of HAE in a human patient who is suffering from an HAE attack. Such a patient can be identified by routine medical procedures. An effective amount of one or more of the provided compounds can be given to the human patient via a suitable route, for example, those described herein. The compounds described herein may be used alone, or may be used in combination with other anti -HAE agents, for example but not limited to, a Cl esterase inhibitor (e.g., Cinryze^® or Berinert^®), a pKal inhibitor (e.g., ecallantide or lanadelumab) or a bradykinin B2 receptor antagonist (e.g., Firazyr^®).

[0062] In other embodiments, provided herein are methods or reducing the risk of

HAE attack in a human HAE patient who is in quiescent stage. Such a patient can be identified based on various factors, including history of HAE attack. An effective amount of one or more of the compounds can be given to the human patient via a suitable route, for example, those described herein. The compounds described herein may be used alone, or may be used in combination with other anti -HAE agents, for example but not limited to, a Cl esterase inhibitor (e.g., Cinryze^® or Berinert^®), a pKal inhibitor (e.g., ecallantide or lanadelumab) or a bradykinin B2 receptor antagonist (e.g., Firazyr^®).

[0063] In yet other embodiments, provided herein are prophylactic treatment of HAE in human patients having risk to HAE attacks with one or more of the compounds described herein. Patients suitable for such prophylactic treatment may be human subjects having history of HAE attacks (e.g., human subjects experiencing more than 2 attacks per month). Alternatively, patients suitable for the prophylactic treatment may be human subjects having no HAE attack history but bearing one or more risk factors for HAE (e.g., family history, genetic defects in Cl-INH gene, etc.) Such prophylactic treatment may involve the compounds described herein as the sole active agent, or involve additional anti-HAE agents, such as those described herein.

[0064] In certain embodiments, provided herein are methods for preventing or reducing edema in an eye of a subject (e.g. , a human patient). In some examples, the human patient is a diabetic having, suspected of having, or at risk for diabetic macular edema (DME). DME is the proliferative form of diabetic retinopathy characterized by swelling of the retinal layers, neovascularization, vascular leak, and retinal thickening in diabetes mellitus due to leaking of fluid from blood vessels within the macula. In certain embodiments of practicing this method, an effective amount of one or more of the compounds described herein, or pharmaceutically acceptable salts thereof, may be delivered into the eye of the subject where treatment is needed. For example, the compound may be delivered by intraocular injection, or intravitreal injection. A subject may be treated with the compound as described herein, either as the sole active agent, or in combination with another treatment for DME. Non-limiting examples of treatment for DME include laser photocoagulation, steroids, VEGF pathway targeting agents (e.g., Lucentis® (ranibizumab) or Eylea^® (aflibercept)), and/or anti-PDGF agents.

[0065] In certain embodiments, the methods disclosed herein comprise administering to the subject an effective amount of Compound 6. In some embodiments, the effective amount is a therapeutically effective amount. In some embodiments, the effective amount is a prophylactically effective amount.

[0066] In certain embodiments, the subject being treated is an animal. The animal may be of either sex and may be at any stage of development. In certain embodiments, the subject is a mammal. In certain embodiments, the subject being treated is a human. In certain embodiments, the subject is a domesticated animal, such as a dog, cat, cow, pig, horse, sheep, or goat. In certain embodiments, the subject is a companion animal, such as a dog or cat. In certain embodiments, the subject is a livestock animal, such as a cow, pig, horse, sheep, or goat. In certain embodiments, the subject is a zoo animal. In another embodiment, the subject is a research animal such as a rodent (e.g., mouse, rat), dog, pig, or non-human primate. In certain embodiments, the animal is a genetically engineered animal.

In certain embodiments, the animal is a transgenic animal.

[0067] Certain methods described herein may comprise administering one or more additional pharmaceutical agent(s) in combination with the compounds described herein. The additional pharmaceutical agent(s) may be administered at the same time as Compound 6, or at different times than Compound 6. For example, Compound 6 and any additional pharmaceutical agent(s) may be on the same dosing schedule or different dosing schedules. All or some doses of the Compound 6 may be administered before all or some doses of an additional pharmaceutical agent, after all or some does an additional pharmaceutical agent, within a dosing schedule of an additional pharmaceutical agent, or a combination thereof.

The timing of administration of Compound 6 and additional pharmaceutical agents may be different for different additional pharmaceutical agents. [0068] In certain embodiments, the additional pharmaceutical agent comprises an agent useful in the treatment of an edema, such as HAE or DME. Examples of such agents are provided herein.

Assays

[0069] To develop useful pKal inhibitors, candidate inhibitors capable of decreasing pKal activity may be identified in vitro. The activity of the inhibitor compounds can be assayed utilizing methods known in the art and/or those methods presented herein.

Pharmaceutical Compositions

[0070] In another aspect, the present invention provides pharmaceutical compositions comprising any of the compounds described herein (e.g., Compound 6) or any of the compounds described herein (e.g., Compound 6) in combination with a pharmaceutically acceptable excipient (e.g., carrier).

[0071] The pharmaceutical compositions include optical isomers, diastereomers, or pharmaceutically acceptable salts of the inhibitors disclosed herein.

[0072] A “pharmaceutically acceptable carrier,” as used herein refers to pharmaceutical excipients, for example, pharmaceutically, physiologically, acceptable organic or inorganic carrier substances suitable for enteral or parenteral application that do not deleteriously react with the active agent. Suitable pharmaceutically acceptable carriers include water, salt solutions (such as Ringer's solution), alcohols, oils, lipids, gelatins, and carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethy cellulose, and polyvinyl pyrrolidine. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with the compounds of the invention.

[0073] The compounds of the invention can be administered alone or can be coadministered to the subject. Coadministration is meant to include simultaneous or sequential administration of the compounds individually or in combination (more than one compound). The preparations can also be combined, when desired, with other active substances (e.g. to reduce metabolic degradation). Formulations

[0074] Compounds of the present invention can be prepared and administered in a wide variety of oral, parenteral, and topical dosage forms. Thus, the compounds of the present invention can be administered by injection (e.g. intravenously, intramuscularly, intracutaneously, subcutaneously, intraduodenally, or intraperitoneally). Also, the compounds described herein can be administered by inhalation, for example, intranasally. Additionally, the compounds of the present invention can be administered transdermally. It is also envisioned that multiple routes of administration (e.g., intramuscular, oral, transdermal) can be used to administer the compounds of the invention. Accordingly, the present invention also provides pharmaceutical compositions comprising a pharmaceutically acceptable carrier or excipient and one or more compounds of the invention.

[0075] For preparing pharmaceutical compositions from the compounds of the present invention, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier can be one or more substance that may also act as diluents, flavoring agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material.

[0076] In powders, the carrier is a finely divided solid in a mixture with the finely divided active component. In tablets, the active component is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.

[0077] The powders and tablets preferably contain from 5% to 70% of the active compound. Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. The term “preparation” is intended to include the formulation of the active compound with encapsulating material as a carrier providing a capsule in which the active component with or without other carriers, is surrounded by a carrier, which is thus in association with it. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

[0078] For preparing suppositories, a low melting wax, such as a mixture of fatty acid glycerides or cocoa butter, is first melted and the active component is dispersed homogeneously therein, as by stirring. The molten homogeneous mixture is then poured into convenient sized molds, allowed to cool, and thereby to solidify.

[0079] Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water/propylene glycol solutions. For parenteral injection, liquid preparations can be formulated in solution in aqueous polyethylene glycol solution.

[0080] When parenteral application is needed or desired, particularly suitable admixtures for the compounds of the invention are injectable, sterile solutions, preferably oily or aqueous solutions, as well as suspensions, emulsions, or implants, including suppositories. In particular, carriers for parenteral administration include aqueous solutions of dextrose, saline, pure water, ethanol, glycerol, propylene glycol, peanut oil, sesame oil, polyoxyethylene-block polymers, and the like. Ampoules are convenient unit dosages. The compounds of the invention can also be incorporated into liposomes or administered via transdermal pumps or patches. Pharmaceutical admixtures suitable for use in the present invention include those described, for example, in Pharmaceutical Sciences (17th Ed., Mack Pub. Co., Easton, PA) and WO 96/05309.

[0081] Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizers, and thickening agents as desired. Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known suspending agents.

[0082] Also included are solid form preparations that are intended to be converted, shortly before use, to liquid form preparations for oral administration. Such liquid forms include solutions, suspensions, and emulsions. These preparations may contain, in addition to the active component, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like.

[0083] The pharmaceutical preparation is preferably in unit dosage form. In such form the preparation is subdivided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules. Also, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form. [0084] The quantity of active component in a unit dose preparation may be varied or adjusted according to the particular application and the potency of the active component. The composition can, if desired, also contain other compatible therapeutic agents.

Examples

[0085] The examples below are meant to illustrate certain embodiments of the invention, and not to limit the scope of the invention.

General Experimental

[0086] Abbreviations

Ή NMR Proton Nuclear Magnetic Resonance API Active Pharmaceutical Ingredient ASR Analytical Service Report ca. Approximately CCD Charge Coupled Detector DCM Dichloromethane DMSO Dimethyl sulfoxide DSC Differential Scanning Calorimetry DVS Dynamic Vapour Sorption eq Equivalents

EtOH Ethanol

GVS Gravimetric Vapour Sorption

H2O Water

HPLC High Performance Liquid Chromatography

IC Ion Chromatography

ID Identification

IDR Intrinsic Dissolution Rate IP Intellectual Property

IPA 2-Propanol

IPAC Isopropyl Acetate

KF Karl Fischer

MDSC Modulated Differential Scanning Calorimetry

MeCN Acetonitrile

MEK Methyl ethyl ketone

MeOH Methanol

MIBK Methyl isobutyl ketone

MSZW Metastable Zone Width

N/A Not Applicable

NMP /V-Methylpyrrolidone

NMR Nuclear Magnetic Resonance

PLM Polarised Light Microscopy

RH Relative Humidity

RM Reaction Mixture

RT Room Temperature

SCXRD Single Crystal X-Ray Diffraction

TBME /CT/-Butyl methyl ether t-BuOH /e/V-Butanol

TFA Trifluoroacetic acid

TGA Thermal Gravimetric Analysis

THF Tetrahydrofuran

UV Ultraviolet

VH-XRPD Variable Humidity X-Ray Powder Diffraction vol Volumes

VT-XRPD Variable Temperature X-Ray Powder Diffraction XRPD X-Ray Powder Diffraction

Instruments and Methods A. X-rav powder diffraction (XRPD)

[0087] For XRPD analysis, the following X-ray powder diffractometers were used.

The parameters used are listed in Table 5. Unless otherwise stated, 2-theta (2Q) values disclosed herein were obtained using a Bruker AXS D8 Advance with parameters described in Table 5.

Table 5. XRPD Parameters

B Thermogravimetric (TGA) and Differential Scanning Calorimetry (DSC)

[0088] TGA data were collected using a TA Q500/Q5000 TGA from TA Instruments.

DSC was performed using a TA Q200/Q2000 DSC from TA Instruments. Detailed parameters used are listed in Table 6.

Table 6. TGA and DSC Parameters

C. HPLC

Agilent 1100 HPLC was utilized and detailed chromatographic conditions are listed in Table 7.

Table 7. Chromatographic conditions and parameters

D. Solution NMR

[0089] Solution NMR was collected on Bruker 400M NMR Spectrometer using

DMSO-rie unless otherwise stated.

E. Intrinsic Dissolution Rate (IDR)

[0090] About 40 mg of the sample was compressed in a 6 mm disc recess, under 100 kg for 2 minutes, with greaseproof paper on the compression base, to form nondisintegrating discs. The discs were then plugged with a bung so that only one surface was exposed to the media during analysis and transferred to the Sirius inForm dissolution apparatus. Analysis was performed at 37 °C in 40 mL media (36 mL ISA water, 4 mL 0.1 M Acetate Phosphate Buffer). Dissolution data were collected over 4 pH sectors (pH 2.0, 5.5, 6.5, and 7.4) for a total of 2 hours - 30 minutes per sector, with UV spectra collected every 30 seconds. A stir speed of 225 rpm was used with a 10 mm path length probe. The IDR was calculated based on the surface area of the 3/6/8 mm disc recess used (28.3 mm2 surface area). XRPD analysis was performed on all samples, both after compression of the material into the disc recess, and post dissolution analysis to observe any change in form. XRPD diffractograms were collected on the Bruker C2.

Example 1: N-((7-chloro-8-fluoroimidazo[l,5-a]pyridin-l-yl)methyl)-l-((6- cyclopropylimidazo[l,2-a]pyridin-2-yl)methyl)-lH-l,2,3-triazole-4-carboxamide (Compound 1)

[0091] The synthesis of Compound 1 is described in detail at Example 148 of the

‘897 application, which is reproduced herein for ease of reference.

Synthesis of 5-cyclopropylpyridin-2-amine.

3 3, ₂ 95 °C, 14 h

[0092] A mixture of 5-bromopyridin-2-amine (100 g, 585 mmol), cyclopropylboronic acid (60 g, 701 mmol), Pd(AcO)2 (6.5 g, 29 mmol), SPhos (24 g, 58.5 mmol) and K3PO4 (372 g, 1.755 mol) in toluene/PhO (1.2 L/0.12 L) was stirred at 90 °C for 14 h under N2. The reaction was concentrated in vacuo to give the crude, which was purified with silica gel chromatography (PE/EA = 1/2) to give the 5-cyclopropylpyridin-2-amine (61 g, yield: 78%) as ayellow solid. ESI-MS [M+H]⁺: 135.1.

Synthesis of 2-(chloromethyl)-6-cyclopropylimidazo[l,2-a]pyridine

[0093] A mixture of 5-cyclopropylpyridin-2-amine (61 g, 455 mmol) and 1,3- dichloropropan-2-one (172 g, 1365 mmol) in EtOH (1 L) was stirred at 95 °C for 13 h. The reaction was concentrated to remove the EtOH. The pH of the residue was adjusted to 9 by addition of aqueous NaHCCb and extracted with EtOAc (1 L x 3). The combined organic layers were washed with brine, dried over Na2SC>4 and concentrated in vacuo to give the crude, which was purified with silica gel chromatography (EA) to give the 2-(chloromethyl)- 6-cyclopropylimidazo[l,2-a]pyridine (40 g, yield: 42%) as a yellow solid. ESI-MS [M+H]⁺: 207.1.

Synthesis of 2-(azidomethyl)-6-cyclopropylimidazo[l,2-a]pyridine.

[0094] To a solution of 2-(chloromethyl)-6-cyclopropylimidazo[l,2-a]pyridine (40 g,

193 mmol) in DMF (600 mL) was added NaN3 (18.8 g, 290 mmol). The resulting reaction was stirred at RT for 2 h. The reaction was diluted with EhO (500 mL) and extracted with EtOAc (500 mL x 3). The combined organic layers were washed with brine, dried over Na2S04 and concentrated in vacuo to give the crude, which was purified with silica gel chromatography (PE/EA = 2/1) to give the 2-(azidomethyl)-6-cyclopropylimidazo[l,2- ajpyridine (35 g, yield: 85%) as a yellow solid. ESI-MS [M+H]⁺: 214.1.

Synthesis of ethyl l-((6-cyclopropylimidazo[l,2-a]pyridin-2-yl)methyl)-lH-l,2,3- triazole-4-carboxylate.

[0095] A mixture of 2-(azidomethyl)-6-cyclopropylimidazo[l,2-a]pyridine (35 g,

163.5 mmol), ethyl propiolate (17.6 g, 180 mmol), CuS04 (2.6 g, 16.35 mmol) and sodium ascorbate (3.3 g, 16.35 mmol) in EhO/t-BuOH (150 mL/150 mL) was stirred at RT for 3 h. Yellow solid was precipitated after 3 h and the mixture was filtered. The cake was dried to give the ethyl l-((6-cyclopropylimidazo[l,2-a]pyridin-2-yl)methyl)-lH-l,2,3-triazole-4- carboxylate (29 g, yield: 57%) as a yellow solid, which was used in the next step without further purification. ESI-MS [M+H]⁺: 312.1.

Synthesis of l-((6-cyclopropylimidazo[l,2-a]pyridin-2-yl)methyl)-lH-l,2,3-triazole-4- carboxylic acid.

[0096] A mixture of ethyl l-((6-cyclopropylimidazo[l,2-a]pyridin-2-yl)methyl)-lH- l,2,3-triazole-4-carboxylate (29 g, 93.2 mmol) and LiOH (6.7 g, 279.6 mmol, solution in 50 mL H2O) in THF/EtOH (150 mL/150 mL) was stirred at 50 °C for 2 h. The reaction was concentrated to remove most of the solvent. The pH of the residue was adjusted to 4 by 2 N HC1 and a pink solid was precipitated out. The mixture was filtered and the filter cake was dried to give l-((6-cyclopropylimidazo[l,2-a]pyridin-2-yl)methyl)-lH-l,2,3-triazole-4- carboxylic acid (20 g, 77%) as a pink solid. ESI-MS [M+H]⁺: 284.1.

Synthesis of N-((7-chloro-8-fluoroimidazo[l,5-a]pyridin-l-yl)methyl)-l-((6- cyclopropylimidazo[l,2-a]pyridin-2-yl)methyl)-lH-l,2,3-triazole-4-carboxamide.

[0097] To a suspension of l-((6-cyclopropylimidazo[l,2-a]pyridin-2-yl)methyl)-lH- l,2,3-triazole-4-carboxylic acid (37 mg, 0.13 mmol) and (7-chloro-8-fluoroimidazo[l,5- a]pyridin-l-yl)methanamine hydrochloride (35 mg, 0.15 mmol) in DMF (3 mL) was added HOBT (40 mg, 0.3 mmol) and EDCI (57 mg, 0.3 mmol), followed by DIPEA (65 mg, 0.5 mmol). The resulting mixture was stirred at RT for 12 h. The reaction mixture was poured into H2O (15 mL) slowly. The suspension mixture was stirred for 1 h, and filtered. The filtered cake was washed with H2O (20 mL) and MeOH (20 mL) then dried under vacuum pump to give the N-((7-chloro-8-fluoroimidazo[l,5-a]pyridin-l-yl)methyl)-l-((6- cyclopropylimidazo[l,2-a]pyridin-2-yl)methyl)-lH-l,2,3-triazole-4-carboxamide as pale solid (30 mg, yield: 50%). ESI-MS [M+H]⁺: 465.0. Purity: 98.4% (214 nm), 98.5% (254 nm). ¾ NMR (400 MHz, DMSO-de): 8.70 (t, J = 5.4 Hz, 1H), 8.55 (s, 1H), 8.44 (d, J = 2.4 Hz, 1H), 8.35 (s, 1H), 8.21 (d, J = 7.4 Hz, 1H), 7.83 (s, 1H), 7.41 (d, J = 9.3 Hz, 1H), 7.01 (dd, J = 9.4, 1.8 Hz, 1H), 6.76 (dd, J = 7.3, 6.6 Hz, 1H), 5.73 (s, 2H), 4.70 (d, J = 5.5 Hz, 2H), 1.94- 1.90 (m, 1H), 0.94-0.89 (m, 2H), 0.69-0.65 (m, 2H). Example 2: Compound 6 (oxalate salt)

Compound 6

Compound 6 Pattern 1

[0098] Compound 6 Pattern 1 was isolated from experiments in both acetone and ethyl acetate. This material was shown to contain one equivalent of oxalic acid by ion chromatography, and had a single melt in its DSC thermogram with onset at 212.8 °C. The XRPD pattern of the material after storage at conditions of 40 °C/75%RH for one week was slightly changed to that of the material isolated from screening.

[0099] Scale-up : Compound 1 obtained from the synthetic procedure described in

Example 1 was charged to an HEL polyblock 100 mL vessel. Acetone (100 vol, 50 mL) was charged to the vessel and the resulting suspension was heated to 50 °C with 500 rpm suspended magnetic stirring. One equivalent of oxalic acid (1 M in THF) was added to the suspension. The suspension was held isothermally for 30 minutes with stirring then cooled to 5 °C at 0.1 °C/min. The suspension was then held isothermally for 14 hours and the resulting material isolated by filtration and dried under suction. The material was isolated in high yield (95 %) and purity (98.4 %).

Table 8. Characterization of the Scaled-up Compound 6 Pattern 1

[0100] Figure 1 provides the XRPD pattern of the Compound 6 Pattern 1.

[0101] Figure 2 provides the TGA/DSC curves of Compound 6 Pattern 1.

[0102] Thermal analysis of the material showed thermal decomposition of the material to occur at approximately 200 °C, and the DSC trace contained a single endotherm with a shoulder (onset 211.3 °C, 197 J/g). GVS analysis of the material showed it to be slightly hygroscopic with a 0.6 % mass increase observed between 0 and 90 %RH, with no observed change to the material as evidenced by XRPD post analysis. The material was shown to be unchanged by XRPD after storage for one week at conditions of 40 °C/75%RH and 25 °C/97%RH.

Further Experiments of Additional Forms of Compound 6

[0103] 30 mg Compound 1 was charged to 7 mL vials. To these, 100 vol of selected solvents (55 vol used for NMP) was added, and the samples heated to 50 °C (60 °C for DMSO samples as complete dissolution not observed at 50 °C). All samples were stirred with 500 rpm magnetic stirring. Samples were stirred for 30 minutes, then 1 equivalent of the oxalic acid was added (1 M in THF). Samples were then stirred for three hours isothermally and then cooled to 5 °C at 0.1 °C/min (25 °C for DMSO samples). Samples were held isothermally for 16 hours then isolated by filtration. Samples with NMP as solvent were observed to be clear solutions. To these, 55 vol TBME was added, and the samples stirred for a further 30 minutes prior to being isolated by filtration. Solids were analyzed by XRPD, then dried in a vacuum oven overnight at 40 °C before a second analysis by XRPD. Experiment number = 11 -20.

Table 9. Results of the Polymorphism Assessment of the Oxalate Salt

Compound 6 Pattern 4

[0104] As noted above, Compound 6 Pattern 2 was isolated from the screening experiments in IP A. About 10 mg of Compound 6 Pattern 2 was then heated to 175 °C at a rate of 10 °C in a TGA pan, held isothermally for 5 minutes, then cooled to ambient temperature to obtain Compound 6 Pattern 4. The TGA thermogram of the material, which showed no mass loss corresponding to desolvation demonstrated the material to be a solvent- free form. The TGA thermogram was seen to contain a mass loss between 200 and 260 °C corresponding to 0.84 molar equivalents of oxalic acid. The temperature of this mass loss coincides with an endotherm in the DSC trace of the material (onset 223.8 °C, 215 J/g).

[0105] Scale-up : Compound 1 obtained from the synthetic procedure described in

Example 1 was charged to an HEL polyblock 140 mL vessel. To this, 100 vol (50 mL) IPA was added and the sample heated to 50 °C and held isothermally for 30 minutes with 500 rpm stirring using an HEL poly block. One equivalent oxalic acid was added (1 M in THF), the sample was stirred for 3 hours, and then cooled to 5 °C at 0.1 °C/min. The sample was then stirred for 3 days, with aliquots removed intermittently and analysed by XRPD. After 3 days, the sample was adjudged to have fully converted and was therefore isolated by filtration and dried under vacuum. This sample was heated to 175 °C for 5 minutes using a Karl Fischer oven, then analysed by XRPD. Table 10. Characterization of the Scaled -up Compound 6 Pattern 4

[0106] Figure 3 provides the XRPD pattern of the Compound 6 Pattern 4.

[0107] Figure 4 provides the TGA/DSC curves of Compound 6 Pattern 4.

[0108] Compound 6 Pattern 4 was shown to be slightly hygroscopic by GVS with a

0.66 % mass loss observed between 0 and 90 %RH. Analysis of the material post GVS analysis showed it to have been unchanged as demonstrated by XRPD. The material was shown to be unchanged by XRPD after storage for one week at conditions of 40 °C/75 %RH and 25 °C/97%RH.

[0109] Alternatively, Compound 6 Pattern 4 can be prepared according to the following procedure: Compound 1 obtained from the synthetic procedure described in Example 1 (500 mg) was charged to a 100 mL round bottom flask. To this, 50 vol (25 mL) MeCNiEhO 1:1 was added and the suspension heated to 50 °C. To this, two equivalents of oxalic acid (1 M in THF) were added and clarification of the suspension was observed. The solution was stirred isothermally for 30 minutes, then cooled to 5 °C at 0.1 °C/min. The resulting suspension was held isothermally for 12 hours, then isolated by filtration and dried under suction. The 523.19 mg material was recovered with a yield of 87.6%. The material was shown to have an XRPD pattern consistent with Oxalate Salt - Pattern 4. Ion chromatography showed the material to contain 1.0 equivalent of oxalic acid, demonstrating that although two equivalents of oxalic acid were used, the material is a monosalt.

Competitive Slurry between Compound 6 Pattern 1 and Compound 6 Pattern 1

[0110] 120 mg of both Compound 6 Pattern 1 and Compound 6 Pattern 4 were dispensed into a 20 mL vial. These were mixed, and then aliquots (20 mg) of this mixture was dispensed into HPLC vials and 90 vol selected solvents were added. Samples were slurried using a Polar Bear with 500 rpm magnetic stirring. Aliquots were removed after four days and analyzed by XRPD.

[0111] The results of the competitive slurries of Compound 6 Pattern 1 and

Compound 6 Pattern 4 are shown in Table 11. In all but one experiment (Experiment 1), after four days of slurrying, the sole detectable phase by XRPD was Compound 6 Pattern 4.

In the remaining experiment in which ethanol was used as a solvent and the material was slurried at 5 °C the XRPD pattern of the isolated material was shown to be a mixture of Compound 6 Pattern 4 and a novel pattern, hypothesized to be an ethanol solvate. Taken together, the competitive slurry data demonstrates that Compound 6 Pattern 4 is the more thermodynamically stable of the two solvent-free oxalate salts.

Table 11. Results of Competitive Slurry of Compound 6 Pattern 4 and Compound 6 Pattern 1

Example 3: Solid Form Solubility and Scalability

[0112] The tabulated IDR sector 1 average values of the oxalate solid forms and reference compound solid forms are shown in Table 12. It is evident from the IDR data, that three stable solid forms (oxalate, Compound 6 Pattern 1 and Pattern 4; succinate, Compound 4 Pattern 1; and L-malate, Compound 3 Pattern 1) exhibit increased sector 1 IDR compared to free base form Compound 1 Form 1. One solid form, Compound 4 Pattern 1, fumarate, exhibit decreased sector 1 IDR compared to free base form Compound 1 Form 1. From these data it can be seen that the L-malate solid form (Compound 3 Form 1) has the highest IDR. However, no solid materials of L-malate, Pattern 1 could be isolated during solution-based scale-up experiments. In addition, further solution-based scale up experiments for succinate showed that external HC1 is required for solution clarification (dissolution of all materials), making it a less attractive candidate for scale up production.

[0113] Compound 6 Pattern 4 is more thermodynamically stable than Compound 6

Pattern 1 in competitive slurry experiments. A solution-based and operationally simple scalable crystallization procedure for the isolation of phase pure Compound 6 Pattern 4 in high yield, purity and low residual solvent content was identified.

Table 12. Sector 1 IDR Results of Scaled-up Solid Forms and Free Base Material of Compound 1

Reference Example 1: Compound 1 Form 1 (free base)

[0114] Compound 1 Form 1 can be prepared from an amorphous form of Compound 1.

Acetone (100 vol, 42 mL) was charged to the vessel and the amorphous form was slurried at 5 °C with 500 rpm suspended magnetic stirring for 3 days. The solid was subsequently isolated by filtration and dried under suction for 30 minutes. Recovery: 339.33 mg, Yield: 80.2%.

Table 13. Characterisation of Compound 1 Form 1

Reference Example 2: Compound 3 (L-malate salt)

Compound 3 Pattern 1

[0115] Compound 1 obtained from the synthetic procedure described in Example 1 was charged to an HEL polyblock 100 mL vial. Ethyl acetate (100 vol, 50 mL) was subsequently charged to the vial and the resulting suspension was heated to 50 °C with 500 rpm suspended magnetic stirring. To this, one equivalent L-malic was added (1 M in THF). The suspension was held isothermally for 16 hours with stirring, during which time, aliquots were removed and analyzed by XRPD. After 16 hours, conversion was not observed to have occurred. Suspension then cooled to 5 °C at 0.5 °C/min and held isothermally for 72 hours then a further aliquot was isolated and analyzed by XRPD. Conversion was observed to have occurred. The resulting solid was isolated by filtration and dried under suction. The material was isolated in 78.4 % yield with a purity of 98.2 %.

Table 14. Characterization of the Scaled-up Compound 3 Pattern 1

Reference Example 3: Compound 4 (succinate salt)

Compound 4

Compound 4 Pattern 1

[0116] Compound 1 obtained from the synthetic procedure described in Example 1 was charged to an HEL polyblock 100 mL vial. Acetone (100 vol, 50 mL) was subsequently charged to the vial and the resulting suspension was heated to 50 °C with 500 rpm suspended magnetic stirring. To this, one equivalent succinic acid (1 M in MeOH) was added to give a suspension. The suspension was held isothermally for 16 hours with stirring, and aliquots were removed and analyzed by XRPD. After 16 hours, conversion was observed to have occurred. Suspensions were then cooled to 5 °C at 0.5 °C/min, held isothermally for 1 hour then isolated by filtration. The material was isolated in 80.5 % yield with a purity of 98.2 %.

Table 15. Characterization of the Scaled-up Compound 4 Pattern 1

Claims

CLAIMS We Claim:

1. A solid form of Compound 1 :

1 selected from the group consisting of a Compound 1 salt or co-crystal form of oxalic acid.

2. The solid form of claim 1, wherein the solid form is crystalline.

3. A solid form of Compound 6, comprising Compound 1 and oxalic acid:

6

4. The solid form of claim 3, where the solid form is Compound 6 Pattern 1 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising three or more diffractions at angles (2 theta ± 0.2) of 4.7, 6.1, 18.7, 24.0, 24.2, and 24.6.

5. The solid form of claim 3, where the solid form is Compound 6 Pattern 1 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising four or more diffractions at angles (2 theta ± 0.2) of 4.7, 6.1, 18.7, 24.0, 24.2, and 24.6.

6. The solid form of claim 3, where the solid form is Compound 6 Pattern 1 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising five or more diffractions at angles (2 theta ± 0.2) of 4.7, 6.1, 18.7, 24.0, 24.2, and 24.6.

7. The solid form of claim 3, where the solid form is Compound 6 Pattern 1 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising diffractions at angles (2 theta ± 0.2) of 4.7, 6.1, 18.7, 24.0, 24.2, and 24.6.

8. The solid form of claim 3, where the solid form is Compound 6 Pattern 1 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising diffractions at angles (2 theta ± 0.2) of 4.7, 6.1, 18.7, 24.0, 24.2, and 24.6, corresponding to d-spacing (angstroms ± 0.2) of 18.88, 14.36, 4.73, 3.71, 3.67, and 3.62 (respectively).

9. The solid form of claim 3, where the solid form is Compound 6 Pattern 1 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising diffractions at angles (2 theta ± 0.2) of:

10. The solid form of claim 3, where the solid form is Compound 6 Pattern 1 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising diffractions at angles (2 theta ± 0.2) corresponding to d-spacing (angstroms ± 0.2) of:

11. The solid form of any one of claims 3-10, characterized by a differential scanning calorimetry (DSC) endotherm having a minima at about 211.3 °C.

12. The solid form of any one of claims 3-11, characterized by an about 0.6% mass increase between 0 and 90%RH by gravimetric vapor sorption (GVS) analysis.

13. The solid form of claim 3, where the solid form is Compound 6 Pattern 4 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising three or more diffractions at angles (2 theta ± 0.2) of 6.4, 11.9, 19.2, 26.1, 26.6, and 27.2.

14. The solid form of claim 3, where the solid form is Compound 6 Pattern 4 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising four or more diffractions at angles (2 theta ± 0.2) of 6.4, 11.9, 19.2, 26.1, 26.6, and 27.2.

15. The solid form of claim 3, where the solid form is Compound 6 Pattern 4 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising five or more diffractions at angles (2 theta ± 0.2) of 6.4, 11.9, 19.2, 26.1, 26.6, and 27.2.

16. The solid form of claim 3, where the solid form is Compound 6 Pattern 4 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising diffractions at angles (2 theta ± 0.2) of 6.4, 11.9, 19.2, 26.1, 26.6, and 27.2.

17. The solid form of claim 3, where the solid form is Compound 6 Pattern 4 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising diffractions at angles (2 theta ± 0.2) of 6.4, 11.9, 19.2, 26.1, 26.6, and 27.2, corresponding to d-spacing (angstroms ± 0.2) of 13.89, 7.43, 4.63, 3.42, 3.35, and 3.27 (respectively).

18. The solid form of claim 3, where the solid form is Compound 6 Pattern 4 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising diffractions at angles (2 theta ± 0.2) of:

19. The solid form of claim 3, where the solid form is Compound 6 Pattern 4 and is characterized by an X-ray powder diffraction (XRPD) pattern comprising diffractions at angles (2 theta ± 0.2) corresponding to d-spacing (angstroms ± 0.2) of:

20. The solid form of any one of claims 3 and 13-19, characterized by a differential scanning calorimetry (DSC) endotherm having a minima at about 222.9 °C.

21. The solid form of any one of claims 3 and 13-20, characterized by an about 0.66% mass increase between 0 and 90%RH by gravimetric vapor sorption (GVS) analysis.

22. A solid form of Compound 6:

6 prepared by a method comprising the steps of combining Compound 1 and oxalic acid in IP A, and then removing the solvent to provide the solid form of Compound 6.

23. The solid form of claim 22, prepared by a method comprising the steps of adding a solution of oxalic acid in THF to a solution of Compound 1 in IP A, and then removing the solvent to provide the solid form of Compound 6.

24. The solid form of claim 22, prepared by a method comprising the steps of: i) adding a solution of oxalic acid in THF to a solution of Compound 1 in IPA at 50 °C, ii) cooling the combined solution of step i) to 5 °C, iii) stirring the cooled solution for a time sufficient to form the solid form of Compound 6, and iv) filtering the combined solution to remove the solvent.

25. A solid form of Compound 6:

6 prepared by a method comprising the steps of combining Compound 1 and oxalic acid in acetone, and then removing the solvent to provide the solid form of Compound 6.

26. The solid form of claim 25, prepared by a method comprising the steps of adding a solution of oxalic acid in THF to a solution of Compound 1 in acetone, and then removing the solvent to provide the solid form of Compound 6.

27. The solid form of claim 25, prepared by a method comprising the steps of: v) adding a solution of oxalic acid in THF to a solution of Compound 1 in acetone at 50 °C, vi) cooling the combined solution of step i) to 5 °C, vii) stirring the cooled solution for a time sufficient to form the solid form of Compound 6, and viii) filtering the combined solution to remove the solvent.

28. A pharmaceutical composition comprising the solid form according to any one of claims 1-27 and a pharmaceutical acceptable carrier or excipient.

29. A method of preparing a solute form of Compound 6,

6 wherein the method includes the formation of Compound 6 Pattern 1.

30. A method of preparing a solid form of Compound 6,

6 wherein the method includes the formation of Compound 6 Pattern 4.

31. A method of preparing a solid form of Compound 6,

6 wherein the method does not include the formation of Compound 6 Pattern 1 or Compound 6 Pattern 4.

32. A method of treating a plasma kallikrein-mediated disease or disorder using a solid form or composition of any one of the preceding claims.

33. A method of treating hereditary angioedema or diabetic macular edema comprising administering to a patient in need thereof a solid form or composition of any one of the preceding claims.