New solid form of a plasma kallikrein inhibitor

Abstract

The invention provides a new polymorph of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide, a screening method for detecting it, pharmaceutical compositions containing it and its use in therapy.

Description

[0001] NEW SOLID FORM OF A PLASMA KALLIKREIN INHIBITOR

[0002] The present invention relates to a new solid form of a plasma kallikrein inhibitor, pharmaceutical compositions containing it, and its use in therapy. Also provided are processes for preparing the solid form of the present invention. The present invention also relates to a screening method for detecting the new solid form in a sample.

[0003] BACKGROUND TO THE INVENTION

[0004] Inhibitors of plasma kallikrein have a number of therapeutic applications, particularly in the treatment of bradykinin mediated angioedema such as hereditary angioedema and bradykinin mediated angioedema non-hereditary (BK-AEnH).

[0005] Plasma kallikrein is a trypsin-like serine protease that can liberate kinins from kininogens (see K. D. Bhoola et al., "Kallikrein-Kinin Cascade", Encyclopedia of Respiratory Medicine, p483- 493; J. W. Bryant et al., "Human plasma kail ikrein-kinin system: physiological and biochemical parameters" Cardiovascular and haematological agents in medicinal chemistry, 7, p234-250, 2009; K. D. Bhoola et al., Pharmacological Rev., 1992, 44, 1 ; and D. J. Campbell, "Towards understanding the kallikrein-kinin system: insights from the measurement of kinin peptides", Brazilian Journal of Medical and Biological Research 2000, 33, 665-677). It is an essential member of the intrinsic blood coagulation cascade, although its role in this cascade does not involve the release of bradykinin or enzymatic cleavage. Plasma prekallikrein is encoded by a single gene and can be synthesized in the liver, as well as other tissues. It is secreted by hepatocytes as an inactive plasma prekallikrein that circulates in plasma as a heterodimer complex bound to high molecular weight kininogen (HK) which is activated to give the active plasma kallikrein. This contact activation system (or contact system) can be activated by negatively charged surfaces that activate Factor XII (FXII) to Factor XI la (FXIIa), by certain proteases e.g. plasmin (Hofman et al., Clin Rev Allergy Immunol 2016), which may not require negative surfaces, or by misfolded proteins (Maas et al., J Clinical Invest 2008). FXIIa mediates conversion of plasma prekallikrein to plasma kallikrein and the subsequent cleavage of high molecular weight kininogen (HK) to generate bradykinin, a potent inflammatory hormone. Kinins are potent mediators of inflammation that act through G protein-coupled receptors and antagonists of kinins (such as bradykinin receptor antagonists) have previously been investigated as potential therapeutic agents for the treatment of a number of disorders (F. Marceau and D. Regoli, Nature Rev., Drug Discovery, 2004, 3, 845-852). Plasma kallikrein is thought to play a role in a number of inflammatory disorders. The major inhibitor of plasma kallikrein is the serpin C1 esterase inhibitor. Patients who present with a genetic deficiency in C1 esterase inhibitor suffer from hereditary angioedema (HAE) which results in intermittent swelling of face, hands, throat, gastro-intestinal tract and genitals. Blisters formed during acute episodes contain high levels of plasma kallikrein which cleaves high molecular weight kininogen (HK) liberating bradykinin leading to increased vascular permeability. Treatment with a large protein plasma kallikrein inhibitor has been shown to effectively treat HAE by preventing the release of bradykinin which causes increased vascular permeability (A. Lehmann "Ecallantide (DX-88), a plasma kallikrein inhibitor for the treatment of hereditary angioedema and the prevention of blood loss in on-pump cardiothoracic surgery" Expert Opin. Biol. Ther. 8, p1187-99). However, risks of anaphylactic reactions have been reported for Ecallantide.

[0006] Hereditary angioedema is a rare inherited disorder characterised by recurrent acute attacks where fluids accumulate outside of the blood vessels, blocking the normal flow of blood or lymphatic fluid and causing rapid swelling of tissues such as in the hands, feet, limbs, face, intestinal tract, or airway. “Hereditary angioedema” can thus be defined as any disorder characterised by recurrent episodes of bradykinin mediated angioedema (e.g. severe swelling) caused by an inherited dysfunction / fault / mutation. There are currently three known categories of HAE: (i) HAE type 1 , (ii) HAE type 2, and (iii) normal C1 inhibitor HAE (normal C1 Inh HAE). However, the HAE field is developing quickly and further types of HAE might be defined in the future.

[0007] Without wishing to be bound by theory, it is thought that HAE type 1 is caused by mutations in the SERPING1 gene that lead to reduced levels of C1 inhibitor in the blood. Without wishing to be bound by theory, it is thought that HAE type 2 is caused by mutations in the SERPING1 gene that lead to dysfunction of the C1 inhibitor in the blood. Without wishing to be bound by theory, the cause of normal C1 Inh HAE is less well defined and the underlying genetic dysfunction / fault / mutation can sometimes remain unknown. What is known is that the cause of normal C1 Inh HAE is not related to reduced levels or dysfunction of the C1 inhibitor (in contrast to HAE types 1 and 2). Normal C1 Inh HAE can be diagnosed by reviewing the family history and noting that angioedema has been inherited from a previous generation (and thus it is hereditary angioedema). Normal C1 Inh HAE can also be diagnosed by determining that there is a dysfunction / fault / mutation in a gene other than those related to C1 inhibitor. For example, it has been reported that dysfunction / fault / mutation with plasminogen can cause normal C1 Inh HAE (see e.g. Veronez et al., Front Med (Lausanne). 2019 Feb 21 ;6:28. doi: 10.3389 / fmed.2019.00028; or Recke et al., Clin Transl Allergy. 2019 Feb 14;9:9. doi: 10.1186 / s13601-019-0247-x.). It has also been reported that dysfunction / fault / mutation with Factor XII can cause normal C1 Inh HAE (see e.g. Mansi et al., The Association for the Publication of the Journal of Internal Medicine Journal of Internal Medicine, 2015, 277; 585- 593. doi: 10.1111 / joim.12304; or Maat et al. J Thromb Haemost. 2019 Jan; 17(1):183-194. doi: 10.1111 / jth.14325).

[0008] Acute HAE attacks normally progress through three key clinically distinct stages: an initial prodromal stage (that can typically last for up to 12 hours), followed by a swelling stage, and then an absorption stage. A majority of HAE attacks announce themselves with prodromal symptoms. Two thirds of prodromes appeared less than 6 hours before a HAE attack and no prodromes occur more than 24 hours before a HAE attack (Magerl et al., Clinical and Experimental Dermatology 2014, 39, 298-303). For example, the following prodromal symptoms may start to be observed: a slight swelling (particularly affecting the face and neck), a typical type of abdominal pain, a typical reddening of the skin called "erythema marginatum". An attack is fully developed when it has reached maximum swelling and maximum expression of pain (e.g. abdominal attack), discomfort (e.g. peripheral attack) or threat to life (e.g. laryngeal attack). Once the attack has reached its peak, the subsequent time period to normalization is determined by the time it takes for the swelling to disappear and the liquid that has penetrated the tissues to be reabsorbed.

[0009] However, angioedemas are not necessarily inherited. Indeed, another class of angioedema is bradykinin mediated angioedema non-hereditary (BK-AEnH), which is not caused by an inherited genetic dysfunction / fault / mutation. Often the underlying cause of BK-AEnH is unknown and / or undefined. However, the signs and symptoms of BK-AEnH are similar to those of HAE, which without being bound by theory, is thought to be on account of the shared bradykinin mediated pathway between HAE and BK-AEnH. Specifically, BK-AEnH is characterised by recurrent acute attacks where fluids accumulate outside of the blood vessels, blocking the normal flow of blood or lymphatic fluid and causing rapid swelling of tissues such as in the hands, feet, limbs, face, intestinal tract, airway or genitals. BK-AEnH attacks are acute and normally progress through three key clinically distinct stages: an initial prodromal stage (that can typically last for up to 12 hours), followed by a swelling stage, and then an absorption stage. A majority of attacks announce themselves with prodromal symptoms. Two thirds of prodromes appeared less than 6 hours before an attack and no prodromes occur more than 24 hours before an attack (Magerl et al. Clinical and Experimental Dermatology (2014) 39, pp298-303). For example, the following prodromal symptoms may start to be observed: a slight swelling (particularly affecting the face and neck), a typical type of abdominal pain, a typical reddening of the skin called "erythema marginatum". An attack is fully developed when it has reached maximum swelling and maximum expression of pain (e.g. abdominal attack), discomfort (e.g. peripheral attack) or threat to life (e.g. laryngeal attack). Once the attack has reached its peak, the subsequent time period to normalization is determined by the time it takes for the swelling to disappear and the liquid that has penetrated the tissues to be reabsorbed.

[0010] Specific types of BK-AEnH include: non hereditary angioedema with normal C1 Inhibitor (AE- nC1 Inh), which can be environmental, hormonal, or drug induced; acquired angioedema; anaphylaxis associated angioedema; angiotensin converting enzyme (ACE) inhibitor induced angioedema; dipeptidyl peptidase 4 inhibitor induced angioedema; and tPA induced angioedema (tissue plasminogen activator induced angioedema). However, reasons why these factors and conditions cause angioedema in only a relatively small proportion of individuals are unknown.

[0011] Environmental factors that can induce AE-nC1 Inh include air pollution (Kedarisetty et al., Otolaryngol Head Neck Surg. 2019 Apr 30: 194599819846446. doi: 10.1177 / 0194599819846446) and silver nanoparticles such as those used as antibacterial components in healthcare, biomedical and consumer products (Long et al., Nanotoxicology. 2016;10(4):501-11. doi: 10.3109 / 17435390.2015.1088589).

[0012] Various publications suggest a link between the bradykinin and contact system pathways and BK-AEnHs, and also the potential efficacy of treatments, see e.g.: Bas et al. (N Engl J Med 2015); Leibfried and Kovary. (J Pharm Pract 2017); van den Elzen et al. (Clinic Rev Allerg Immunol 2018); Han et al. (JCI 2002). tPA induced angioedema is discussed in various publications as being a potentially life threatening complication following thrombolytic therapy in acute stroke victims (see e.g. Simao et al., Blood. 2017 Apr 20; 129(16):2280-2290. doi: 10.1182 / blood-2016-09-740670; Frohlich et al., Stroke. 2019 Jun 11 :STROKEAHA119025260. doi: 10.1161 / STROKEAHA.119.025260; Rathbun, Oxf Med Case Reports. 2019 Jan 24;2019(1):omy112. doi: 10.1093 / omcr / omy112; Lekoubou et al., Neurol Res. 2014 Jul;36(7):687-94. doi: 10.1179 / 1743132813Y.0000000302; Hill et al., Neurology. 2003 May 13;60(9):1525-7).

[0013] Stone et al. (Immunol Allergy Clin North Am. 2017 Aug;37(3):483-495.) reports that certain drugs can cause angioedema. Scott et al. (Curr Diabetes Rev. 2018;14(4):327-333. doi: 10.2174 / 1573399813666170214113856) reports cases of dipeptidyl peptidase-4 inhibitor induced angioedema.

[0014] Hermanrud et al. (BMJ Case Rep. 2017 Jan 10;2017. pii: bcr2016217802) reports recurrent angioedema associated with pharmacological inhibition of dipeptidyl peptidase 4 and also discusses acquired angioedema related to angiotensin-converting enzyme inhibitors (ACEI- AAE). Kim et al. (Basic Clin Pharmacol Toxicol. 2019 Jan;124(1):115-122. doi: 10.1111 / bcpt.13097) reports angiotensin II receptor blocker (ARB)-related angioedema. Reichman et al., (Pharmacoepidemiol Drug Saf. 2017 Oct;26(10):1190-1196. doi: 10.1002 / pds.4260) also reports angioedema risk for patients taking ACE inhibitors, ARB inhibitors and beta blockers. Diestro et al. (J Stroke Cerebrovasc Dis. 2019 May;28(5):e44- e45. doi: 10.1016 / j.jstrokecerebrovasdis.2019.01.030) also reports a possible association between certain angioedemas and ARBs.

[0015] Giard et al. (Dermatology. 2012;225(1):62-9. doi: 10.1159 / 000340029) reports that bradykinin mediated angioedema can be precipitated by oestrogen contraception.

[0016] Synthetic and small molecule plasma kallikrein inhibitors have been described previously, for example by Garrett et al. ("Peptide aldehyde...." J. Peptide Res. 52, 62-71 (1998)), T. Griesbacher et al. ("Involvement of tissue kallikrein but not plasma kallikrein in the development of symptoms mediated by endogenous kinins in acute pancreatitis in rats" British Journal of Pharmacology 137, 692-700 (2002)), Evans ("Selective dipeptide inhibitors of kallikrein" WO03 / 076458), Szelke et al. ("Kininogenase inhibitors" WO92 / 04371), D. M. Evans et al. (Immunolpharmacology, 32, p115-116 (1996)), Szelke et al. ("Kininogen inhibitors" WO95 / 07921), Antonsson et al. ("New peptides derivatives" WO94 / 29335), J. Corte et al. (’’Six membered heterocycles useful as serine protease inhibitors” W02005 / 123680), J. Sturzbecher et al. (Brazilian J. Med. Biol. Res 27, p1929-34 (1994)), Kettner et al. (US 5, 187,157), N. Teno et al. (Chem. Pharm. Bull. 41 , p1079-1090 (1993)), W. B. Young et al. ("Small molecule inhibitors of plasma kallikrein" Bioorg. Med. Chem. Letts. 16, p2034-2036 (2006)), Okada et al. ("Development of potent and selective plasmin and plasma kallikrein inhibitors and studies on the structure-activity relationship" Chem. Pharm. Bull. 48, p1964-72 (2000)), Steinmetzer et al. ("Trypsin-like serine protease inhibitors and their preparation and use" WO08 / 049595), Zhang et al. ("Discovery of highly potent small molecule kallikrein inhibitors" Medicinal Chemistry 2, p545-553 (2006)), Sinha et al. ("Inhibitors of plasma kallikrein" W008 / 016883), Shigenaga et al. (“Plasma Kallikrein Inhibitors” WO2011 / 118672), and Kolte et al. (“Biochemical characterization of a novel high-affinity and specific kallikrein inhibitor”, British Journal of Pharmacology (2011), 162(7), 1639-1649). Also, Steinmetzer et al. (“Serine protease inhibitors” WO2012 / 004678) describes cyclized peptide analogs which are inhibitors of human plasmin and plasma kallikrein.

[0017] As explained above, HAE can manifest in patients who present with a genetic deficiency or dysfunction in C1 esterase inhibitor. Thus, some of the current treatments of HAE involve administering a C1 esterase inhibitor to normalise the deficiency or dysfunction in C1 esterase inhibitor. Such treatments can be prophylactic (i.e. administered in the absence of acute HAE attack symptoms to prevent / reduce the likelihood of an acute HAE attack) and / or acute treatments (i.e. administered when acute HAE attack symptoms are noticed to try to stop or reduce the severity of the acute HAE attack).

[0018] Cinryze® and Haegarda® contain a C1 esterase inhibitor and are authorised to prevent acute HAE attacks (i.e. prophylactic treatment). Treatment with Cinryze® requires the preparation of a solution from a powder, which is then injected every 3 or 4 days. Similarly, treatment with Haegarda® requires the preparation of a solution from a powder, which is then injected twice a week. It is not always possible for a patient to self-administer these treatments, and if this is the case, the patient is required to visit a clinic for treatment. Thus, both of these prophylactic treatments suffer from high patient burden. Additionally, the FDA packet insert for Haegarda® states that it “should not be used to treat an acute HAE attack”, and therefore a patient may require additional therapy if a HAE attack develops.

[0019] Berinert® and Ruconest® contain a C1 esterase inhibitor and are authorised to treat acute HAE attacks. Both of these treatments also involve the preparation of an injectable solution followed by injection. This process can be burdensome on the patient, especially when the patient is suffering from an acute HAE attack. Self-administration of the dosage amount is also not always possible, and if it is not, administration of the drug can be substantially delayed thus increasing the severity of the acute HAE attack for the patient.

[0020] Kalbitor® (active substance ecallantide) and Takhzyro® (active substance lanadelumab) are selective plasma kallikrein inhibitors approved for medical use in the treatment of HAE. Both treatments are formulated as solutions for injection. Ecallantide is a large protein plasma kallikrein inhibitor that presents a risk of anaphylactic reactions. Indeed, the Ell marketing authorisation application for Kalbitor® has recently been withdrawn because the benefits of Kalbitor® are said to not outweigh its risks. Lanadelumab is a recombinant fully human IgG 1 kappa light chain monoclonal antibody. Reported adverse reactions of treatment with lanadelumab include hypersensitivity, injection site pain, injection site erythema, and injection site bruising. The authorised EMA label for Takhzyro® (active substance lanadelumab) states that it “is not intended for treatment of acute HAE attacks” and that “in case of a breakthrough HAE attack, individualized treatment should be initiated with an approved rescue medication”. Also, as injections, both of these treatments involve a high patient burden.

[0021] Berotralstat (BCX7353) has been approved in some countries for preventative treatment of HAE (not as an on demand treatment), e.g. under the brand name Orladeyo®. Hwang et al. (Immunotherapy (2019) 11(17), 1439-1444) states that higher doses were associated with more gastrointestinal adverse effects indicating increased toxicity at higher levels.

[0022] Furthermore, many molecules in the known art feature a highly polar and ionisable guanidine or amidine functionality. It is well known that such functionalities may be limiting to gut permeability and therefore to oral availability. For example, it has been reported by Tamie J. Chilcote and Sukanto Sinha (“ASP-634: An Oral Drug Candidate for Diabetic MacularEdema”, ARVO 2012 May 6th - May 9th, 2012, Fort Lauderdale, Florida, Presentation 2240) that ASP- 440, a benzamidine, suffers from poor oral availability. It is further reported that absorption may be improved by creating a prodrug such as ASP-634. However, it is well known that prodrugs can suffer from several drawbacks, for example, poor chemical stability and potential toxicity from the inert carrier or from unexpected metabolites. In another report, indole amides are claimed as compounds that might overcome problems associated with drugs possessing poor or inadequate ADME-tox and physicochemical properties although no inhibition against plasma kallikrein is presented or claimed (Griffioen et al., “Indole amide derivatives and related compounds for use in the treatment of neurodegenerative diseases”, W02010142801).

[0023] Other plasma kallikrein inhibitors known in the art are generally small molecules, some of which include highly polar and ionisable functional groups, such as guanidines or amidines. Recently, plasma kallikrein inhibitors that do not feature guanidine or amidine functionalities have been reported. For example Brandl et al. (“N-((6-amino-pyridin-3-yl)methyl)-heteroaryl- carboxamides as inhibitors of plasma kallikrein” WO2012 / 017020), Evans et al. (“Benzylamine derivatives as inhibitors of plasma kallikrein” WO2013 / 005045), Allan et al. (“Benzylamine derivatives” WO2014 / 108679), Davie et al. (“Heterocyclic derivates” WO2014 / 188211), Davie et al. (“N-((het)arylmethyl)-heteroaryl-carboxamides compounds as plasma kallikrein inhibitors” WO2016 / 083820).

[0024] As noted, hereditary angioedema (HAE) is a rare inherited condition characterised by recurrent attacks of severe swelling. The swelling is typically of the face, hands, throat, gastrointestinal tract and / or genitals. Attacks of swelling can be very unpleasant and dangerous if untreated, sometimes causing abdominal pain, diarrhoea, vomiting and even death. Existing treatments of HAE are either administered on-demand or as a prophylactic. On-demand treatment is administered in response to a specific attack to lessen its severity e.g. at the first sign of attack symptoms. Prophylactic treatment involves continuously and regularly taking treatment to reduce the likelihood of an attack occurring i.e. not in response to one specific attack. Prophylactic treatments are generally not completely effective, with patients receiving prophylactic treatment often suffering from breakthrough attacks. For this reason, it is common for HAE patients on prophylactic treatment to have an on-demand treatment as a backup for such breakthrough attacks (such treatment is often called a “rescue treatment”). Accordingly, all HAE patients, regardless of their current treatment regime (or lack thereof) would benefit from having effective on-demand treatments of HAE, which is a need addressed by the present invention.

[0025] There is therefore a need for a treatment of bradykinin-mediated angioedema (e.g. HAE or BK-AEnH) that is less burdensome on the patient to improve patient compliance. In particular, there is a need for a treatment of bradykinin-mediated angioedema (e.g., HAE or BK-AEnH) that can be administered orally. HAE attacks resolve faster and are shorter after early treatment (Maurer M et al. PLoS ONE 2013;8(2): e53773. doi:10.1371 / journal. pone.0053773) and thus early intervention when an attack is expected, or ongoing, is essential to desirably manage the disease. Therefore, there is also a need for an oral treatment of acute bradykinin-mediated angioedema (e.g. HAE or BK-AEnH) attacks on-demand e.g. upon recognition of symptoms of an acute bradykinin-mediated angioedema (e.g. HAE or BK-AEnH) attack and does not require regular (or continuous) dosing e.g. a treatment that does not require injections twice a week. In particular there is a need for an effective oral treatment in young patients (e.g. between the ages of 2 and less than 18). Existing injectable treatments can cause pain at the injection site, and the injection itself can cause the child stress. Moreover, there is a need for specific oral dosage forms that are more suitable for patients who may struggle to swallow tablets (e.g. children, the elderly, patients suffering from a laryngeal attack or from conditions such as dysphagia).

[0026] To date, there is a lack of approved oral treatments for bradykinin-mediated angioedema (e.g. HAE or BK-AEnH) (see e.g. Craig et al. Int Arch Allergy Immunol. 2014;165(2):119-27. doi: 10.1159 / 000368404; Mager et al., Immunol Allergy Clin North Am. 2017 Aug; 37(3): 571-584). There are no approved oral on-demand treatments for bradykinin-mediated angioedema disorders (e.g. HAE or BK-AEnH). Instead, the majority of available treatments are injectables. Injectables suffer from many disadvantages compared to oral treatments, largely relating to patient compliance. For example, injectables are more inconvenient than oral treatments e.g. needing to be in a suitable environment to formulate and then administer the injection, and also causing pain at the injection site. Reduced patient compliance can lead to patients delaying treatment, which generally leads to attack symptoms getting worse, or even avoiding treatment altogether. Indeed, many patients suffering from HAE simply hope that any attack will dissipate on its own to avoid administering an injection. Injectable treatments suffer from late dosing because the patient may need to prepare the dosage form or even travel to hospital for treatment. Therefore, HAE treatment is often undermined by late dosing caused by the high burden on the patient. Indeed, Maurer M et al. (PLoS ONE 2013;8(2): e53773. doi:10.1371 / journal. pone.0053773) explains that more than 60% of patients administer their HAE injectable more than one hour after the onset of an attack. HAE drugs are often used off-label to treat some types of BK-AEnH, so the problems associated with HAE treatments apply similarly to treatment of BK-AEnH.

[0027] Icatibant (marketed as Firazyr®) is the predominant on-demand treatment for HAE and is administered by subcutaneous injection. In a real-world study of HAE patients with access to icatibant (Maurer M et al. PLoS ONE 2013;8(2): e53773. doi:10.1371 / journal. pone.0053773), less than 40% of treated attacks were dosed within an hour of onset, with 30% not being treated until over 5 hours after onset. Further, 45% of attacks in this study were not treated at all. Therefore, there is an unmet need to provide patients with improved treatment options to treat HAE attacks on-demand.

[0028] As noted above, there is a lack of any approved oral on-demand treatments for bradykinin-mediated angioedema disorders such as HAE and BK-AEnH.

[0029] The applicant has developed a novel series of compounds that are inhibitors of plasma kallikrein, which are disclosed in W02016 / 083820 (PCT / GB2015 / 053615). These compounds demonstrate good selectivity for plasma kallikrein. One such compound is N-[(3-fluoro-4- methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide. Certain dosage forms comprising this compound are disclosed in Collett et al. WO2019 / 106361. The name N-[(3-fluoro-4- methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide denotes the structure of the compound of Formula A:

[0030]

[0031] Formula A

[0032] The compound of Formula A is also known by its codename “KVD900” and its International Nonproprietary Name (INN), “sebetralstat”. Sebetralstat is currently in phase 3 clinical trials (known as the KONFIDENT study, https: / / konfidentstudy.com / ; see Example 16), having met all of its phase 2 efficacy and safety endpoints (see clinical trial NCT04208412; Example 15).

[0033] In the manufacture of pharmaceutical formulations, it is important that the active compound be in a form in which it can be conveniently handled and processed in order to obtain a commercially viable manufacturing process. Accordingly, the chemical stability and the physical stability of the active compound are important factors. The active compound, and formulations containing it, must be capable of being effectively stored over appreciable periods of time, without exhibiting any significant change in the physico-chemical characteristics (e.g. chemical composition, density, hygroscopicity and solubility) of the active compound.

[0034] It is known that manufacturing a particular solid-state form of a pharmaceutical ingredient can affect many aspects of its solid state properties and offer advantages in aspects of solubility, dissolution rate, chemical stability, mechanical properties, technical feasibility, processability, pharmacokinetics and bioavailability. Some of these are described in "Handbook of Pharmaceutical Salts; Properties, Selection and Use", P. Heinrich Stahl, Camille G. Wermuth (Eds.) (Verlag Helvetica Chimica Acta, Zurich). Methods of manufacturing solid-state forms are also described in "Practical Process Research and Development", Neal G. Anderson (Academic Press, San Diego) and "Polymorphism: In the Pharmaceutical Industry", Rolf Hilfiker (Ed) (Wiley VCH). Polymorphism in pharmaceutical crystals is described in Byrn (Byrn, S.R., Pfeiffer, R.R., Stowell, J.G., "Solid-State Chemistry of Drugs", SSCI Inc., West Lafayette, Indiana, 1999), Brittain, H.G., "Polymorphism in Pharmaceutical Solids", Marcel Dekker, Inc., New York, Basel, 1999) or Bernstein (Bernstein, J., "Polymorphism in Molecular Crystals", Oxford University Press, 2002).

[0035] A number of solid forms of the compound of Formula A, and salts thereof, were previously disclosed in the applicant’s patent application, PCT / GB2017 / 051579 (WO2017 / 208005), including:

[0036] • four crystalline polymorphs of the compound of Formula A (labelled as “Form 1”, “Form 2”, “Form 3” and “Form 4”);

[0037] • four crystalline polymorphs of the hydrochloride salt of the compound of Formula A (labelled as “Form 5”, “Form 6”, “Form 7” and “Form 18”);

[0038] • one crystalline polymorph of the sulfate salt of the compound of Formula A (labelled as “Form 8);

[0039] • three crystalline polymorphs of the phosphate salt of the compound of Formula A (labelled as “Form 9”, “Form 10” and “Form 11”);

[0040] • two crystalline polymorphs of the mesylate salt of the compound of Formula A (labelled as “Form 12” and “Form 13”);

[0041] • one crystalline polymorph of the tosylate salt of the compound of Formula A (labelled as “Form 14”);

[0042] • two crystalline polymorphs of the edisylate salt of the compound of Formula A (labelled as “Form 15” and “Form 16”); and

[0043] • one crystalline polymorph of the besylate salt of the compound of Formula A (labelled as “Form 17”).

[0044] The labelling used in PCT / GB2017 / 051579 (WO2017 / 208005) for the known solid forms will be used when referring to the known forms in this application, except for Form 5 which will be referred to herein as “Form 5 (HCI salt)” to distinguish it from a further crystalline polymorph of the compound of Formula A which was labelled as “Form 5” in another of the applicant’s patent applications, PCT / GB2018 / 053443 (WO2019 / 106361). The further sol id form disclosed in WO2019 / 106361 will be referred to herein as “Form 5 (freebase)”.

[0045] As discussed above, different solid forms of the same compound (or salt thereof) may have different properties. To develop a pharmaceutical product, it is important to identify a sufficiently stable and bioavailable solid form of a drug compound. However, identification of all the possible polymorphic forms of a compound is often complex due to, for example, the existence of a large number of polymorphic forms, kinetic barriers, instability, interconversion of polymorphic forms and difficulties in separating concomitantly crystallised polymorphic forms.

[0046] SUMMARY OF THE INVENTION

[0047] The invention relates to a novel polymorphic form of N-[(3-fluoro-4-methoxypyridin-2- yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4- carboxamide which denotes the structure of the compound of Formula A:

[0048] Formula A

[0049] The compound of Formula A is also known by its codename “KVD900” and its International Non-proprietary Name (INN), “sebetralstat”. In the present application, the novel polymorphic form of the compound of Formula A may be referred to as “Form 19”.

[0050] Form 19 of the compound of Formula A has advantageous physio-chemical properties (such as thermodynamic stability). Unexpectedly, Form 19 was not identified in a polymorph screen.

[0051] DEFINITIONS

[0052] The term “solid form” as used herein includes crystalline forms and amorphous forms.

[0053] Where solid forms of the invention can be provided as one or more geometrical, optical, enantiomeric, diastereomeric and tautomeric forms, including but not limited to cis- and transforms, E- and Z-forms, R-, S- and meso-forms, keto-, and enol-forms, unless otherwise stated a reference to a particular compound includes all such isomeric forms, including racemic and other mixtures thereof. Where appropriate such isomers can be separated from their mixtures by the application or adaptation of known methods (e.g. chromatographic techniques and recrystallisation techniques). Where appropriate such isomers can be prepared by the application or adaptation of known methods (e.g. asymmetric synthesis).

[0054] In the present specification, X-ray powder diffraction peaks (expressed in degrees 20) are measured using Cu Ka radiation.

[0055] When used in the context of X-ray powder diffraction peaks, the term "approximately" means that there is an uncertainty in the measurements of the degrees 20 of ± 0.5 (expressed in degrees 20), preferably ± 0.3 (expressed in degrees 20), more preferably ± 0.2 (expressed in degrees 20) or even more preferably ± 0.1 (expressed in degrees 20).

[0056] The term "approximately" can mean that there is an uncertainty in the measurements of the degrees 20 of ± 0.5 (expressed in degrees 20).

[0057] The term "approximately" can mean that there is an uncertainty in the measurements of the degrees 20 of ± 0.3 (expressed in degrees 20).

[0058] The term "approximately" can mean that there is an uncertainty in the measurements of the degrees 20 of ± 0.2 (expressed in degrees 20).

[0059] The term "approximately" can mean that there is an uncertainty in the measurements of the degrees 20 of ± 0.1 (expressed in degrees 20).

[0060] When used the context of infrared spectrum peaks, the term "approximately" means that there is an uncertainty in the measurements of ± 0.5 cm-1, preferably ± 0.3 cm-1, more preferably ± 0.2 cm-1or even more preferably ± 0.1 cm-1.

[0061] When used in the context of quantities or values, the term “about” means ±20%. The term “about” can also mean ±10%. The term “about” can also mean ±5%. For instance, “about 50%” means “40% to 60%” when “about” is ±20%.

[0062] When used in the context of temperature values, the term “about” can mean ±20°C. Alternatively, when used in the context of temperature values, the term “about” can mean ±10°C. Alternatively, when used in the context of temperature values, the term “about” can mean ±5°C. Alternatively, when used in the context of temperature values, the term “about” can mean ±2°C. Alternatively, when used in the context of temperature values, the term “about” can mean ±1°C. Alternatively, when used in the context of temperature values, the term “about” can mean ±0.5°C. For instance, “about 100°C” means “80-120°C”, when “about” is ±20°C.

[0063] As used herein, weight percentage values (wt.%) are relative to the mass of the whole composition. For example, the weight percentage of Form 19 in a composition consisting only of 80 mg of Form 1 and 20 mg of Form 19 would be expressed as a 20 wt.%.

[0064] The X-ray powder diffraction pattern of a solid form may be described herein as "substantially" the same as that depicted in a figure. It will be appreciated that the peaks in X-ray powder diffraction patterns may be slightly shifted in their positions and relative intensities due to various factors known to the skilled person. For example, shifts in peak positions or the relative intensities of the peaks of a pattern can occur because of the equipment used, method of sample preparation, preferred packing and orientations, the radiation source, and method and length of data collection. However, the skilled person will be able to compare the X-ray powder diffraction patterns shown in the figures herein with those of an unknown solid form to confirm the identity of the solid form.

[0065] The skilled person is familiar with techniques for measuring XRPD patterns. In particular, the X-ray powder diffraction pattern of the sample of compound may be recorded according to Method C, as detailed in General Experimental section of this application.

[0066] In the context of the present invention, references herein to "treatment" include references to curative, palliative and prophylactic treatment, unless there are specific indications to the contrary. The terms "therapy", "therapeutic" and "therapeutically" should be construed in the same way.

[0067] The crystalline form of the present invention may be administered alone or in combination with one or more other drugs. Generally, it will be administered as a formulation in association with one or more pharmaceutically acceptable excipients. The term “excipient” is used herein to describe any ingredient other than the compound(s) of the invention which may impart either a functional (i.e., drug release rate controlling) and / or a non-functional (i.e. , processing aid or diluent) characteristic to the formulations. The choice of excipient will to a large extent depend on factors such as the particular mode of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form.

[0068] References to the compound of Formula A include all isotopic variants. DETAILED DESCRIPTION OF THE INVENTION

[0069] New solid form of the compound of Formula A

[0070] The present invention provides a solid form of the free base of the compound of Formula A. More specifically, the present invention provides a crystalline solid form of the free base of the compound of Formula A, referred to herein as “Form 19”.

[0071] The present invention provides a crystalline form (Form 19) of N-[(3-fluoro-4-methoxypyridin- 2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4- carboxamide, which exhibits one or more of the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately 5.8, 10.1 , 13.1 , 15.0, 16.1 , 17.3, 17.6, 18.8, 19.6, 20.9, 21.8, 22.4 and 23.0. In some embodiments, Form 19 exhibits two or more, three or more, four or more, or five or more of the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately 5.8, 10.1, 13.1 , 15.0, 16.1 , 17.3, 17.6, 18.8, 19.6, 20.9, 21.8, 22.4 and 23.0.

[0072] Preferably, the crystalline form (Form 19) of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3- (methoxymethyl)-1-({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide, exhibits at least the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately: a) 5.8, 10.1 , 15.0, 16.1 and 17.3; or b) 5.8, 10.1 , 13.1 , 15.0, 16.1 , 17.3 and 17.6; or c) 5.8, 10.1 , 13.1 , 15.0, 16.1 , 17.3, 17.6, 18.8 and 19.6; or d) 5.8, 10.1 , 13.1 , 15.0, 16.1 , 17.3, 17.6, 18.8, 19.6, 20.9, 21.8, 22.4; or e) 5.8, 10.1 , 13.1 , 15.0, 16.1 , 17.3, 17.6, 18.8, 19.6, 20.9, 21.8, 22.4 and 23.0.

[0073] More preferably, Form 19 exhibits at least the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately 5.8, 10.1 , 15.0, 16.1 and 17.3.

[0074] The present invention also provides a crystalline solid form (Form 19) of the compound of Formula A having an X-ray powder diffraction pattern substantially the same as that show in Figure 12. The skilled person is familiar with techniques for obtaining XRPD patterns. The XRPD pattern of a sample of the compound of Formula A may be obtained according to a method outlined in the General Experimental section of this application, in particular Method C.

[0075] The present invention also provides a crystalline solid form (Form 19) of the compound of Formula A, which exhibits an endothermic peak in its DSC thermograph at 162.59 °C. Form 19 may have an endothermic peak in its DSC thermograph at 163 ± 3°C.

[0076] The present invention also provides a crystalline solid form (Form 19) of the compound of Formula A, which has a DSC thermograph substantially the same as that shown in Figure 13.

[0077] The skilled person will be familiar with techniques for measuring Differential Scanning Calorimetry (DSC) thermographs. The DSC thermograph of a sample of the compound of Formula A may be recorded according to a method outlined in the General Experimental section of this application, in particular Method B.

[0078] The present invention also provides a crystalline solid form (Form 19) of the compound of Formula A, which has an IR spectrum substantially the same as that shown in Figure 32.

[0079] Preferably, the crystalline form (Form 19) of the compound of Formula A exhibits at least the following characteristic peaks (expressed in cm-1) in its IR spectrum at approximately:

[0080] (i) 1562.5; or

[0081] (ii) 1562.5 and 1657.8; or

[0082] (iii) Each of the peaks listed in Table 12.

[0083] Preferably, the crystalline form (Form 19) of the compound of Formula A does not exhibit a characteristic peak (expressed in cm-1) in its IR spectrum at approximately 1639.5.

[0084] Preferably, the crystalline form (Form 19) of the compound of Formula A does not exhibit a characteristic peak (expressed in cm-1) in its IR spectrum at approximately 1639.5 and does exhibit at least the following characteristic peaks (expressed in cm-1) in its IR spectrum at approximately: a-1. 1562.5; or b-1. 1562.5 and 1657.8; or c-1. Each of the peaks listed in Table 12. The present invention provides both unsolvated and solvated forms. The term 'solvate' is used herein to describe a molecular complex comprising the compound of Formula A and an amount of one or more pharmaceutically acceptable solvents, for example, ethanol. The term 'hydrate' is employed when the solvent is water. The present invention provides a solvate of the solid form of the compound of Formula A (Form 19). The present invention also provides a solid form of the hydrate of the compound of Formula A (Form 19). The hydrate can be the monohydrate of the compound of Formula A. The hydrate can be the dihydrate of the compound of Formula A.

[0085] In another aspect, the invention provides a screening method for determining whether Form 19 is present in a sample comprising any form of the compound of Formula A (e.g., Form 1).

[0086] Process for making solid form of the invention

[0087] The present invention also encompasses a process for the preparation of Form 19, said process comprising crystallisation of said solid form from a solution of the compound of Formula A in a solvent or mixture of solvents.

[0088] In some embodiments, the solution of the compound of Formula A may be prepared by dissolving a solid form (e.g., Form 1) of the compound of Formula A in a solvent or mixture of solvents.

[0089] In some embodiments, the solution of the compound of Formula A may be prepared by adding a solid form (e.g., Form 1) of the compound of Formula A to a solvent or mixture of solvents to form a suspension and then heating the suspension to dissolve (or partly dissolve) the solid form of the compound of Formula A. In some embodiments, the suspension is heated to reflux.

[0090] Preferably, the solvent is isopropanol. Preferably, the mixture of solvents comprises isopropanol.

[0091] In some embodiments, Form 19 crystallises from the solution of the compound of Formula A when it is held at an elevated temperature (e.g., above room temperature, which is about 25°C) for a defined period. In some embodiments, the elevated temperature is more than about 30°C, more than about 40°C, more than about 50°C, more than about 60°C, more than about 70°C, more than about 80°C or more than about 90°C. In some embodiments, the elevated temperature is less than about 100°C, less than about 90°C, less than about 80°C, less than about 70°C, less than about 60°C or less than about 50°C. In some embodiments, the elevated temperature is in the range of about 60-100°C, more preferably in the range of about 70-90°C, more preferably in the range of about 75-85°C, most preferably in the range of about 78-83°C.

[0092] In some embodiments, the elevated temperature is about the boiling temperature of the solvent or mixture of solvents. The boiling temperature of a solvent or mixture of solvents is the temperature that the pressure exerted by the vapour of the solvent or mixture of solvents equals the pressure exerted by the surroundings on the solvent or mixture of solvents. As such, the boiling temperature of a solvent or mixture of solvents is a function of the pressure on the solution during crystallisation. Therefore, references to boiling temperature herein encompass deviations from the reported boiling temperature of a solvent or mixture of solvents under standard ambient pressure (1 atm) in embodiments where the crystallisation is performed under a pressure deviating from standard ambient pressure. In some embodiments, the crystallisation of Form 19 from the solution of the compound of Formula A is performed under standard ambient pressure (1 atm). For example, when the solvent is isopropanol, the elevated temperature may be about the boiling temperature of isopropanol under standard ambient pressure (1 atm), i.e., about 82.5°C. In some embodiments where the elevated temperature is about the boiling temperature of the solvent or mixture of solvents, the crystallisation is carried out under reflux.

[0093] In some embodiments, after crystallisation at an elevated temperature, the resulting suspension may be cooled, for example to a temperature less than about 50°C, less than about 40°C, less than about 30°C, less than about 20°C, less than about 10°C or less than about 0°C. In some embodiments the resulting suspension is cooled to a temperature of about -80°C to about 50°C, about -80°C to about 25°C, preferably about -20°C to about 25°C, more preferably about 0°C to about 25°C, most preferably about 0°C to about 5°C.

[0094] Compositions comprising the solid form of the invention

[0095] The invention also provides compositions comprising Form 19 of the compound of Formula A (as described herein), wherein the composition is substantially free of amorphous compound of Formula A. As used herein, the term “substantially free of amorphous compound of Formula A” means that the composition contains no significant amount of amorphous compound of Formula A. At least about 90% by weight of the composition can be crystalline solid form. At least about 95% by weight of the composition can be crystalline solid form. More specifically, at least about 97%, 98% or 99% by weight of the composition can be crystalline solid form. The composition can contain less than about 50% by weight of amorphous compound of Formula A. The composition can contain less than about 30% by weight of amorphous compound of Formula A. The composition can contain less than about 20% by weight of amorphous compound of Formula A. The composition can contain less than about 10% by weight of amorphous compound of Formula A. The composition can contain less than about 5% by weight of amorphous compound of Formula A. The composition can contain less than about 3%, 2% or 1% by weight of amorphous compound of Formula A.

[0096] The invention also provides compositions comprising Form 19 of the compound of Formula A, wherein the composition is substantially free of another solid form of the compound of Formula A, wherein “another solid form of the compound of Formula A” includes amorphous compound of Formula A, all crystalline forms of the compound of Formula A other than Form 19 and all salts of the compound of Formula A. As used herein, the term “substantially free of another solid form of the compound of Formula A” means that the composition contains no significant amount of amorphous compound of Formula A, no significant amount of a crystalline form of the compound of Formula A (other than Form 19) and no significant amount of a salt of the compound of Formula A. At least about 90% by weight of the compound of Formula A in the composition can be Form 19. At least about 95% by weight of the compound of Formula A in the composition can be Form 19. More specifically, at least about 97%, 98% or 99% by weight of the compound of Formula A in the composition can be Form 19. The composition can contain less than about 50% by weight of the compound of Formula A of another solid form of the compound of Formula A. The composition can contain less than about 30% by weight of the compound of Formula A of another solid form of the compound of Formula A. The composition can contain less than about 20% by weight of the compound of Formula A of another solid form of the compound of Formula A. The composition can contain less than about 10% by weight of the compound of Formula A of another solid form of the compound of Formula A. The composition can contain less than about 5% by weight of the compound of Formula A of another solid form of the compound of Formula A. The composition can contain less than about 3%, 2% or 1 % by weight of the compound of Formula A of another solid form of the compound of Formula A.

[0097] Pharmaceutical compositions suitable for the delivery of the crystalline form of the present invention and methods for their preparation will be readily apparent to those skilled in the art. Such compositions and methods for their preparation may be found, for example, in Remington’s Pharmaceutical Sciences, 19th Edition (Mack Publishing Company, 1995).

[0098] For administration to human patients, the total daily dose of the crystalline form of the invention is typically in the range 0.1 mg and 10,000 mg, or between 1 mg and 5000 mg, or between 10 mg and 1000 mg depending, of course, on the mode of administration. If administered by intra- vitreal injection a lower dose of between 0.0001 mg (0.1 pg) and 0.2 mg (200 pg) per eye is envisaged, or between 0.0005 mg (0.5 pg) and 0.05 mg (50 pg) per eye.

[0099] The total daily dose may be administered in single or divided doses and may, at the physician's discretion, fall outside of the typical range given herein. These dosages are based on an average human subject having a weight of about 60kg to 70kg. The physician will readily be able to determine doses for subjects whose weight falls outside this range, such as infants and the elderly.

[0100] Accordingly, the present invention provides a pharmaceutical composition comprising Form 19 of the compound of Formula A, and a pharmaceutically acceptable carrier, diluent or excipient.

[0101] The pharmaceutical compositions may be administered topically (e.g. to the eye, to the skin or to the lung and / or airways) in the form, e.g., of eye-drops, creams, solutions, suspensions, heptafluoroalkane (HFA) aerosols and dry powder formulations; or systemically, e.g. by oral administration in the form of tablets, capsules, syrups, powders or granules; or by parenteral administration in the form of solutions or suspensions; or by subcutaneous administration; or by rectal administration in the form of suppositories; or transdermally. In a further embodiment, the pharmaceutical composition is in the form of a suspension, tablet, capsule, powder, granule or suppository.

[0102] In an embodiment of the invention, the active ingredient is administered orally. Oral administration may involve swallowing, so that the compound enters the gastrointestinal tract, and / or buccal, lingual, or sublingual administration by which the compound enters the blood stream directly from the mouth.

[0103] Formulations suitable for oral administration include solid plugs, solid microparticulates, semisolid and liquid (including multiple phases or dispersed systems) such as tablets; soft or hard capsules containing multi- or nano-particulates, liquids, emulsions or powders; lozenges (including liquid-filled); chews; gels; fast dispersing dosage forms; films; ovules; sprays; and buccal / mucoadhesive patches.

[0104] Formulations suitable for oral administration may also be designed to deliver the crystalline form in an immediate release manner or in a rate-sustaining manner, wherein the release profile can be delayed, pulsed, controlled, sustained, or delayed and sustained or modified in such a manner which optimises the therapeutic efficacy of the said crystalline form. Means to deliver compounds in a rate-sustaining manner are known in the art and include slow release polymers that can be formulated with the said compounds to control their release.

[0105] Liquid (including multiple phases and dispersed systems) formulations include emulsions, suspensions, solutions, syrups and elixirs. Such formulations may be presented as fillers in soft or hard capsules. Liquid formulations may also be prepared by the reconstitution of a solid, for example, from a sachet.

[0106] The crystalline form of the invention may also be used in fast-dissolving, fast-disintegrating dosage forms such as those described in Liang and Chen, Expert Opinion in Therapeutic Patents, 2001, 11 (6), 981-986.

[0107] The formulation of tablets is discussed in Pharmaceutical Dosage Forms: Tablets, Vol. 1, by H. Lieberman and L. Lachman (Marcel Dekker, New York, 1980).

[0108] Screening Method

[0109] The invention provides a screening method for determining whether Form 19 is present in a sample comprising any form of the compound of Formula A (e.g., Form 1 of the compound of Formula A).

[0110] In some embodiments, the screening method comprises the steps of: a. Providing the sample; b. Measuring a property of the sample; and c. Determining whether Form 19 is present in the sample by comparing the result of step b with one or more known results of measurements of the same property for one or more known solid forms of the compound of Formula A.

[0111] In some embodiments, the screening method comprises the steps of: a. Providing the sample; b. Measuring a property of the sample; and c. Determining whether Form 19 is present in the sample by comparing the result of step b with a known result of a measurement of the same property for one or more known solid forms of the compound of Formula A. In some embodiments, determining whether Form 19 is present in the sample means determining that Form 19 is present in the sample. In some embodiments, determining whether Form 19 is present in the sample means determining that Form 19 is not present in the sample.

[0112] As will be appreciated by the skilled person, determining whether Form 19 is present in the sample depends on the detection limit of the method used to measure a property of the sample (i.e., the minimum quantity of Form 19 which must be present in the sample to be detected by the measurement method). Therefore, determining that Form 19 is present in the sample means that Form 19 is present in a quantity equal to or greater than the detection limit of the method used to measure a property of the sample. Similarly, determining that Form 19 is not present in the sample means that Form 19 is absent or present in an amount less than the detection limit of the method used to measure a property of the sample.

[0113] In some embodiments, the sample is a composition comprising the compound of Formula A. In some embodiments, the sample consists of, or consists essentially of, the compound of Formula A. In some embodiments, the sample comprises, or consists essentially of, Form 1 of the compound of Formula A. In some embodiments, the sample comprises, or consists essentially of, Form 1 and Form 19. In some embodiments, the sample is a composition comprising Form 1 of the compound of Formula A and one or more excipients. For example, the composition may be a pharmaceutical composition comprising Form 1 of the compound of Formula A and a pharmaceutically acceptable carrier, diluent or excipient.

[0114] The person skilled in the art is aware of the analytical methods in the art which can be used to determine the chemical or physical properties of a chemical substance or mixture. For a given sample comprising any form of the compound of Formula A, the skilled person knows which analytical methods would be appropriate for determining whether Form 19 is present within the sample, in view of Form 19 being disclosed herein. For example, measuring a property of the sample (or reference sample) may comprise performing one or more of X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), differential thermal analysis (DTA), thermogravimetric analysis (TGA), gravimetric vapour sorption (GVS), simultaneous thermal analysis (STA), infrared (IR) spectroscopy on the sample. Preferably, measuring a property of the sample comprises performing differential scanning calorimetry (DSC) on the sample.

[0115] Non-limiting examples of methodologies for performing some analytical methods (e.g., DSC and XRPD) are provided in the Examples. However, the skilled person is aware of methodologies for analytical methods and would be able to choose and perform an appropriate methodology for investigating a given sample comprising any form of the compound of Formula A. In some embodiments, measuring a property of the sample (or reference sample) comprises performing XRPD on the sample according to Method A, Method B or Method C, as set out in the Examples. In some embodiments, measuring a property of the sample (or reference sample) comprises performing DSC on the sample according to Method A or Method B, as set out in the Examples. In some embodiments, measuring a property of the sample (or reference sample) comprises performing DSC on the sample according to Method A, Method B or Method C, as set out in the Examples. In some embodiments, measuring a property of the sample (or reference sample) comprises performing DSC on the sample according to Method C, as set out in the Examples.

[0116] As used herein, the term “known solid forms of the compound of Formula A” includes:

[0117] • All solid forms of the compound of Formula A, and salts thereof, as reported in WO2017 / 208005 (Forms 1 to 4, Form 5 (HCI salt) and Forms 6 to 18); and

[0118] • The solid form of the compound of Formula A as reported in W02019 / 106361 (i.e. , Form 5 (freebase)); and

[0119] • Form 19 of the compound of Formula A, as defined herein.

[0120] In some embodiments, the one or more known solid forms of the compound of Formula A is Form 1. In some embodiments, the one or more known solid forms of the compound of Formula A is Form 19.

[0121] In some embodiments, the result of step b is compared with a known result of a measurement of the same property of Form 1. In some embodiments, the result of step b is compared with a known result of a measurement of the same property of Form 19. In some embodiments, the result of step b is compared with a known result for a measurement of the same property for a known sample comprising both Form 19 and Form 1. In some embodiments, the result of step b is compared with a known result of a measurement of the same property of Form 19 and a known result of a measurement of the same property of Form 1 .

[0122] In some embodiments, measuring a property of the sample comprises determining the melting temperature of one or more of the components of the sample (e.g., one of more known solid forms of the compound of Formula A present in the sample). Methods for determining the melting temperatures of components in a mixture are known in the art (e.g., DSC). In some embodiments, the result of step b comprises the melting temperatures of one or more of the components of the sample. In some embodiments the one or more known results include the melting temperatures for one or more of the known solid forms of the compound of Formula A (e.g., the melting temperature of Form 1 and / or Form 19).

[0123] In some embodiments, measuring a property of the sample comprises performing DSC on the sample to obtain a DSC thermograph. In some embodiments, the result of step b is a DSC thermograph. Preferably, the result of step b is the temperature values of one or more endothermic peaks in a DSC thermograph of the sample. Preferably, the one or more endothermic peaks correspond to a melting temperature of one or more of the solid forms of the compound of Formula A (e.g., the melting temperature of Form 1 and / or Form 19).

[0124] In some embodiments, the result of step b is compared with one or more DSC thermographs from DSC measurements of one or more known solid forms of the compound of Formula A (e.g., Form 1 and / or Form 19). In some embodiments, the result of step b is compared with a DSC thermograph showing a DSC measurement of Form 1 (e.g., Figure 4 or 33, preferably Figure 33). In some embodiments, the result of step b is compared with a DSC thermograph showing a DSC measurement of Form 19 (e.g., Figure 13 or 34, preferably Figure 34). In some embodiments the result of step b is compared with a DSC thermograph showing a DSC measurement of Form 19 (e.g., Figure 13 or 34, preferably Figure 34) and a DSC thermograph showing a DSC measurement of Form 1 (e.g., Figure 4 or 33, preferably Figure 33). In some embodiments, the result of step b is compared with a DSC thermograph showing a DSC measurement of a composition consisting essentially of Form 1 and Form 19 (e.g., Figure 14 or 35, preferably Figure 35). In some embodiments, the result of step b is compared with one or more of the DSC thermographs shown in Figures 4, 13, 14, 31 , 32, 33, 34 and 35.

[0125] In some embodiments, the one or more known results comprise a DSC thermograph obtained by performing DSC on a reference sample. The reference sample may comprise, consist of, or consist essentially of, Form 1 and / or Form 19, preferably Form 19.

[0126] In some embodiments, the screening method comprises the steps of: a. Providing the sample; b. Measuring a property of the sample, comprising performing DSC on the sample to obtain a DSC thermograph; and c. Determining whether Form 19 is present in the sample by comparing the result of step b with a known result of a measurement of the same property for one or more known solid forms of the compound of Formula A. In some embodiments, the screening method comprises the steps of: a. Providing the sample; b. Measuring a property of the sample, comprising performing DSC on the sample to obtain a DSC thermograph; and c. Determining whether Form 19 is present in the sample by comparing the result of step b with a known result of a measurement of the same property for Form 19 and, optionally, Form 1.

[0127] In some embodiments, the screening method comprises the steps of: a. Providing the sample; b. Measuring a property of the sample, comprising performing DSC on the sample to obtain a DSC thermograph; and c. Determining whether Form 19 is present in the sample by comparing the DSC thermograph of step b with a DSC thermograph obtained by performing DSC on Form 19 and, optionally, Form 1.

[0128] In some embodiments, the screening method comprises the steps of: a. Providing the sample; b. Measuring a property of the sample, comprising performing DSC on the sample to obtain a DSC thermograph; and c. Determining whether Form 19 is present in the sample by comparing the DSC thermograph of step b with a DSC thermograph obtained by performing DSC on Form 1 and / or Form 19, preferably Form 19.

[0129] In some embodiments, determining whether Form 19 present in the sample means determining that Form 19 is not present in the sample by comparing the DSC thermograph of step b with a DSC thermograph obtained by performing DSC on Form 1. In some such embodiments, it is determined that Form 19 is not present in the sample by confirming that the DSC thermograph of step b comprises only peaks corresponding to Form 1 (e.g., a single endothermic melting peak corresponding to Form 1). In such an embodiment, the screening method is for determining whether a sample comprising Form 1 does not also comprise any other solid form of the compound of Formula A (e.g., Form 19).

[0130] In some embodiments, determining whether Form 19 is present in the sample means determining that Form 19 is present in the sample by comparing the DSC thermograph of step b with a DSC thermograph obtained by performing DSC on Form 19. In some such embodiments, it is determined that Form 19 is present in the sample by confirming that the DSC thermograph of step b comprises peaks corresponding to Form 19 (e.g., an endothermic melting peak corresponding to Form 19).

[0131] In some embodiments, determining whether Form 19 is present in the sample means determining that Form 19 is not present in the sample by comparing the DSC thermograph of step b with a DSC thermograph obtained by performing DSC on Form 19. In some such embodiments, it is determined that Form 19 is not present in the sample by confirming that the DSC thermograph of step b does not comprises peaks corresponding to Form 19 (e.g., an endothermic melting peak corresponding to Form 19).

[0132] In some embodiments, determining whether Form 19 is present in the sample means determining that Form 19 is present in a sample consisting essentially of Form 1 by comparing the DSC thermograph of step b with a DSC thermograph obtained by performing DSC on Form 19 and a DSC thermograph obtained by performing DSC on Form 1 (optionally a DSC thermograph obtained by performing DSC on a sample consisting essentially of Form 1 and Form 19). In some such embodiments, it is determined that Form 19 present in the sample by confirming that the DSC thermograph of step b comprises peaks corresponding to Form 19 (e.g., an endothermic melting peak corresponding to Form 19).

[0133] In some embodiments, a DSC thermograph of step b comprises an endothermic peak resulting from melting of the Form 1 content of the sample (i.e., an endothermic melting peak). In some embodiments, the DSC thermograph of step b comprises one or more endothermic melting peaks resulting from components of the sample other than Form 1 (e.g., one or more of the known solid forms of the compound of Formula A). In some embodiments, the DSC thermograph of step b comprises an endothermic melting peak resulting from melting of Form 19 present in the sample.

[0134] In some embodiments, the result of step b comprises a value determined from an endothermic peak present in the DSC thermograph of step b. In some embodiments, the one or more known results comprises a value determined from an endothermic peak present in a DSC thermograph of Form 19. In some embodiments, the result of step b comprises one or more of the onset temperatures (TonSet) for one or more of the endothermic melting peaks present in the DSC thermograph of step b. The onset temperature of an endothermic melting peak may be about the melting temperature of a component of the sample (e.g., Form 1 and / or Form 19). In some embodiments, the one or more known results comprises an onset temperature for an endothermic melting peak in a DSC thermograph obtained for a known solid form of the compound of Formula A (e.g., an endothermic melting peak resulting from melting of the Form 19 content of a reference sample). In some embodiments, the one or more known results comprises about the onset temperature of the endothermic melting peak shown in the DSC thermograph of Figure 13 or 34. In some embodiments, the one or more known results comprises an onset temperature for an endothermic melting peak in a DSC thermograph in the range of 156°C and 161°C. In some embodiments, the one or more known results comprises an onset temperature for an endothermic melting peak in a DSC thermograph about 158°C. In some embodiments, the one or more known results comprises an onset temperature for an endothermic melting peak in a DSC thermograph in the range of 150°C and 155°C. In some embodiments, the one or more known results comprises an onset temperature for an endothermic melting peak in a DSC thermograph about 151°C.

[0135] In some embodiments, it is determined that Form 19 is present in the sample if an endothermic melting peak is present in the DSC thermograph of step b corresponding to Form 19 (e.g., an endothermic peak in the range of 156°C to 161 °C). In some embodiments, it is determined that Form 19 is present in the sample if an endothermic melting peak is present in the DSC thermograph of step b with an onset temperature of about 158°C. In some embodiments, it is determined that Form 19 is not present in the sample if an endothermic melting peak is not present in the DSC thermograph of step b corresponding to Form 19 (e.g., an endothermic peak in the range of 156°C to 161 °C). In some embodiments, it is determined that Form 19 is not present in the sample if an endothermic melting peak is not present in the DSC thermograph of step b with an onset temperature of about 158°C.

[0136] If it is determined that the sample comprises Form 19, the screening method may further comprise determining whether the sample comprises an amount of Form 19 which is greater than, equal to, or less than an amount of Form 19 which is present in a reference sample.

[0137] In some embodiments, if it is determined that Form 19 is present in the sample, the screening method further comprises the steps of: d. providing a reference sample comprising Form 19; e. measuring the same property of the reference sample; f. determining whether the sample comprises an amount of Form 19 which is less than, equal to, or greater than the amount Form 19 present in the reference sample by comparing the result of step b with the result of step e. As will be appreciated, the further steps to determine whether the sample comprises an amount of Form 19 which is greater than, equal to, or less than an amount of Form 19 which is present in a reference sample (i.e., steps d-f) can be performed either after or simultaneously to the steps for determining whether Form 19 is present in the sample (i.e., steps a-c). Steps a-f may be performed in any order or simultaneously, provided that: (i) step a precedes step b; (ii) step b precedes step c; (iii) step d precedes step e; (iv) step e precedes step f; and (v) step c precedes step f. In some embodiments, steps a and d are performed simultaneously. In some embodiments, steps b and e are performed simultaneously. In some embodiments, steps a, b, d and e are performed before steps c and f. In one embodiment, steps a and d are performed simultaneously, then steps b and e are performed simultaneously, then step d is performed and then step f is performed.

[0138] In some embodiments, the reference sample comprises, consists of, or consists essentially of, Form 1 and Form 19. In some embodiments, the reference sample comprises, consists of, or consists essentially of, Form 1 and a known amount of Form 19. In some embodiments, the reference sample consists of, or consists essentially of, Form 1 and a known amount of Form 19. In some embodiments, the known amount of Form 19 is a known amount relative to the amount of Form 1 in the reference sample. In some embodiments, the known amount of Form 19 is a known amount relative to the mass of the reference sample. In some embodiments, the known amount of Form 19 is in the range of about 0.1 wt.% to about 2 wt.%. In some embodiments, the known amount of Form 19 may be at least 0.1 wt.%, at least 0.2 wt.%, at least 0.5 wt.%, at least 1.0 wt.% or at least 2.0 wt.% of the reference sample. In some embodiments, the known amount of Form 19 may be about 0.1 wt.%, about 0.2 wt.%, about 0.5 wt.%, about 1.0 wt.% or about 2.0 wt.% of the reference sample. In a preferred embodiment, the known amount of Form 19 in the reference sample is at least 0.5 wt.%. In a preferred embodiment, the known amount of Form 19 in the reference sample is about 0.5 wt.%. In a preferred embodiment, the reference sample consists or consists essentially of, of about 99.5 wt.% of Form 1 and 0.5 wt.% of Form 19. In these embodiments, the mass of the reference sample that is not Form 19 may be Form 1.

[0139] In some embodiments, measuring a property of the sample or the reference sample comprises multiple measurements. For example, in some embodiments, measuring a property of the sample or the reference sample comprises three measurements.

[0140] In some embodiments, measuring a property of the sample comprises performing DSC on the sample multiple times to obtain multiple DSC thermograph. In some embodiments, the result of step b is one or more DSC thermographs. In some embodiments, measuring a property of the sample comprises performing DSC on the sample three times to obtain three DSC thermographs. In some embodiments, the result of step b comprises one or more endothermic melting peaks in the one of more DSC thermographs obtained for the sample (e.g., the endothermic melting peaks for the Form 1 and Form 19 content of the reference sample).

[0141] In some embodiments, measuring a property of the reference sample comprises performing DSC on the reference sample to obtain multiple DSC thermographs. In some embodiments, the result of step e is one or more DSC thermographs. In some embodiments, the result of step e comprises one or more endothermic melting peaks in the DSC thermographs obtained for the reference sample (e.g., the endothermic melting peaks corresponding to Form 1 and Form 19).

[0142] In some embodiments, the screening method further comprises the steps of: d. providing a reference sample comprising Form 19; e. measuring the same property of the reference sample, comprising performing DSC on the reference sample to obtain a DSC thermograph; f. determining whether the sample comprises an amount of Form 19 which is less than, equal to, or greater than the amount of Form 19 present in the reference sample by the result of step b with the result of step e.

[0143] In some embodiments, the screening method further comprises the steps of: d. providing a reference sample comprising Form 19; e. measuring the same property of the reference sample, comprising performing DSC on the reference sample to obtain a DSC thermograph; f. determining whether the sample comprises an amount of Form 19 which is less than, equal to, or greater than the amount of Form 19 present in the reference sample by comparing the DSC thermograph of step b with the DSC thermograph of step e.

[0144] In some embodiments, comparing the DSC thermograph of step b with the DSC thermograph of step e comprises comparing values determined from the peaks in the one or more DSC thermographs of step b with values determined from the peaks in the DSC thermograph of step e. In some embodiments, comparing the one or more DSC thermographs of step b with the DSC thermograph of step e comprises comparing peak areas determined from the peaks in the one or more DSC thermographs of step b with peak areas determined from the peaks in the DSC thermograph of step e. In some embodiments, comparing the one or more DSC thermographs of step b with the DSC thermograph of step e comprises comparing peak areas determined from the endothermic melting peaks in the one or more DSC thermographs of step b with peak areas determined from the endothermic melting peaks in the DSC thermograph of step e. Methods for determining the peak area of a peak in a DSC thermograph are known to the skilled person. In some embodiments, the endothermic melting peaks in the one or more DSC thermographs of step b and the endothermic melting peaks in the one or more DSC thermographs of step e are the endothermic melting peaks corresponding to Form 1 and Form 19. In some embodiments, the endothermic melting peak in a DSC thermograph corresponding to Form 1 is a peak with an onset temperature in the range of 145°C to 155 °C. In some embodiments, the endothermic melting peak in a DSC thermograph corresponding to Form 1 is a peak with an onset temperature in the range of 150°C to 155°C. In some embodiments, the endothermic melting peak in a DSC thermograph corresponding to Form 1 is a peak with an onset temperature at about 151°C. In some embodiments, the endothermic melting peak in a DSC thermograph corresponding to Form 19 is a peak with an onset temperature in the range of 156°C to 161°C. In some embodiments, the endothermic melting peak in a DSC thermograph corresponding to Form 19 is a peak with an onset temperature at about 158°C.

[0145] In some embodiments, comparing the DSC thermograph of step b with the DSC thermograph of step e comprises comparing melting enthalpies calculated from the endothermic melting peaks in the DSC thermographs. In some embodiments, comparing the DSC thermograph of step b with the DSC thermograph of step e comprises comparing melting enthalpies calculated from the endothermic melting peaks for Form 19 in the DSC thermographs. Methods for determining a melting enthalpy from a DSC thermograph are known to the skilled person. The melting enthalpies may be normalised melting enthalpies. Procedures for normalising a melting enthalpy determined from a DSC thermograph are known to the skilled person. For example, a normalised melting enthalpy may be obtained by dividing the calculated melting enthalpy by the mass of the tested sample.

[0146] In some embodiments, determining whether the sample comprises an amount of Form 19 which is less than, equal to, or greater than the amount of Form 19 present in the reference sample comprises comparing the melting enthalpies calculated from the endothermic melting peaks in the DSC thermographs by calculating the ratio of the melting enthalpies for the Form 1 and Form 19 content of the sample (Rt) and the reference sample (Rr). The ratios may be calculated according to the following formula: Where:

[0147] • HfUSForm 19) is the melting enthalpy for the amount of Form 19 present in the sample or reference sample. The melting enthalpy may be calculated by integrating the endothermic melting peak for the Form 19 content (e.g., the endothermic melting peak with an onset temperature at about 158°C).

[0148] • AHf us(Form 1) is the melting enthalpy for the amount of Form 1 present in the sample or reference sample. The melting enthalpy may be calculated by integrating the endothermic melting peak for the Form 1 content (e.g., the endothermic melting peak with an onset temperature at about 151°C).

[0149] In some embodiments, it is determined that the sample comprises an amount of Form 19 which is less than or equal to the amount of Form 19 in the reference sample if Rt< Rr. In some embodiments, it is determined that the sample comprises an amount of Form 19 which is less than the amount of Form 19 in the reference sample if Rt< Rr. In some embodiments, it is determined that the sample comprises an amount of Form 19 which is greater than or equal to the amount of Form 19 in the reference sample if Rt> Rr. In some embodiments, it is determined that the sample comprises an amount of Form 19 which is greater than the amount of Form 19 in the reference sample if Rt> Rr. In some embodiments, it is determined that the sample comprises an amount of Form 19 which is equal to the amount of Form 19 in the reference sample if Rtis equal to about Rr.

[0150] In some embodiments, it is determined that the sample comprises an amount of Form 19 which is less than or equal to the amount of Form 19 in the reference sample if Rt< (Rrx A), where A is a scaling factor. In some embodiments, it is determined that the sample comprises an amount of Form 19 which is less than the amount of Form 19 in the reference sample if Rt

[0151] < (Rrx A), where A is a scaling factor. The value of A can be any value in the range of 0 < A

[0152] < 1. In some embodiments, the value of A is in a range which is one of: 0.6 to 0.9, 0.6 to 0.85, 0.65 to 0.9, 0.65 to 0.85, 0.65 to 0.8, 0.7 to 0.85 or 0.7 to 0.8. In some embodiments, the value of A is about 0.6, about 0.65, about 0.7, about 0.75, about 0.80, about 0.85, about 0.9 or about 0.95. In a preferred embodiment, the value of A is about 0.75. As will be appreciated by the skilled person, the value of A may be determined according to the calculated error margins or standard deviations for the values of Rtand Rr.

[0153] In some embodiments, where multiple DSC thermographs are obtained for the sample and / or reference sample, Rtand / or Rris calculated for each thermograph (e.g., if N thermographs are obtained, N values of Rtare calculated). In some such embodiments, it is determined that the sample comprises an amount of Form 19 which is greater than or equal to the amount of Form 19 in the reference sample if, for any one of the Rtvalues, Rt> Rr. In some such embodiments, it is determined that the sample comprises an amount of Form 19 which is greater than the amount of Form 19 in the reference sample if, for any one of the Rtvalues, Rt> Rr.

[0154] In some embodiments, if (Rrx A) < Rt< Rr, or if (Rrx A) < Rt< Rr, or if (Rrx A) < Rt< Rr, or if (Rrx A) < Rt< Rr, step b is repeated to obtain one or more further DSC thermographs for the sample. In some embodiments, if (Rrx A) < Rt< Rr, step b is repeated to obtain one or more further DSC thermographs for the sample. In some such embodiments, one or more further Rtvalues are calculated from the one or more further DSC thermographs (e.g., three further Rtvalues). In some such embodiments, it is determined that the sample comprises an amount of Form 19 which is less than or equal to the amount of Form 19 in the reference sample if, for all of the one or more further Rtvalues, Rt< Rr.. In some such embodiments, it is determined that the sample comprises an amount of Form 19 which is less than the amount of Form 19 in the reference sample if, for all of the one or more further Rtvalues, Rt< Rr. In some such embodiments, it is determined that the sample comprises an amount of Form 19 which is greater than or equal to the amount of Form 19 in the reference sample if, for any one of the one or more further Rtvalues, Rt> Rr. In some such embodiments, it is determined that the sample comprises an amount of Form 19 which is greater than the amount of Form 19 in the reference sample if, for any one of the one or more further Rtvalues, Rt> Rr.

[0155] Therapeutic Applications

[0156] The present invention also provides a method of treatment of a disease or condition mediated by plasma kallikrein, said method comprising administering to a mammal in need of such treatment a therapeutically effective amount of Form 19 of the compound of Formula A.

[0157] The present invention also provides Form 19, as defined herein, for use in therapy.

[0158] The present invention also provides Form 19, as defined herein, for use in the treatment of a disease or condition mediated by plasma kallikrein.

[0159] In an aspect, the disease or condition mediated by plasma kallikrein is selected from impaired visual acuity, diabetic retinopathy, retinal vascular permeability associated with diabetic retinopathy, diabetic macular edema, hereditary angioedema, diabetes, pancreatitis, cerebral haemorrhage, nephropathy, cardiomyopathy, neuropathy, inflammatory bowel disease, arthritis, inflammation, septic shock, hypotension, cancer, adult respiratory distress syndrome, disseminated intravascular coagulation, blood coagulation during cardiopulmonary bypass surgery, and bleeding from post-operative surgery. In a preferred embodiment, the disease or condition mediated by plasma kallikrein is diabetic macular edema. In a more preferred embodiment, the disease or condition mediated by plasma kallikrein is hereditary angioedema.

[0160] In another aspect, the disease or condition in which plasma kallikrein activity is implicated is retinal vein occlusion.

[0161] Alternatively, the disease or condition mediated by plasma kallikrein may be selected from retinal vascular permeability associated with diabetic retinopathy, diabetic macular edema and hereditary angioedema. Alternatively, the disease or condition mediated by plasma kallikrein may be retinal vascular permeability associated with diabetic retinopathy or diabetic macular edema. Form 19 may be administered in a form suitable for injection into the ocular region of a patient, in particular, in a form suitable for intra-vitreal injection.

[0162] Combination Therapy

[0163] The crystalline form of the present invention may be administered alone or in combination with one or more other drugs. Generally, it will be administered as a formulation in association with one or more pharmaceutically acceptable excipients. The term “excipient” is used herein to describe any ingredient other than the compound(s) of the invention which may impart either a functional (i.e., drug release rate controlling) and / or a non-functional (i.e. , processing aid or diluent) characteristic to the formulations. The choice of excipient will to a large extent depend on factors such as the particular mode of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form.

[0164] In another aspect, the crystalline form of the present invention may be administered in combination with laser treatment of the retina. The combination of laser therapy with intravitreal injection of an inhibitor of VEGF for the treatment of diabetic macular edema is known (Elman M, Aiello L, Beck R, et al. “Randomized trial evaluating ranibizumab plus prompt or deferred laser or triamcinolone plus prompt laser for diabetic macular edema”. Ophthalmology. 27 April 2010).

[0165] Uses and / or methods involving the solid form of the invention The present invention provides the use of the solid form or composition as described herein, in a method of manufacturing a pharmaceutical composition. The present invention also provides the pharmaceutical composition resulting from, obtainable from, and / or obtained by, this use.

[0166] The present invention also provides a method of manufacturing a pharmaceutical composition comprising mixing a solid form or composition as described herein with one or more pharmaceutically acceptable excipients. The present invention also provides the pharmaceutical composition resulting from, obtainable from, and / or obtained by, this method.

[0167] The present invention also provides a method of processing a solid form or composition as described herein into a medicament. The present invention also provides the medicament resulting from, obtainable from, and / or obtained by, this method.

[0168] EXAMPLES

[0169] Figures

[0170] Figure 1 : X-ray powder diffraction pattern of Form A of N-[(3-fluoro-4-methoxypyridin-

[0171] 2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Example 1).

[0172] Figure 2a: X-ray powder diffraction pattern of Form 1 of N-[(3-fluoro-4-methoxypyridin-2- yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Example 2).

[0173] Figure 2b: X-ray powder diffraction pattern of Form 1 of N-[(3-fluoro-4-methoxypyridin-2- yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Example 3).

[0174] Figure 2c: X-ray powder diffraction pattern of Form 1 of N-[(3-fluoro-4-methoxypyridin-2- yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Example 4).

[0175] Figure 3: STA of Form 1 of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-

[0176] (methoxymethyl)-1-({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4- carboxamide (Example 2). Figure 4: DSC of Form 1 of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-

[0177] (methoxymethyl)-1-({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4- carboxamide (Example 2).

[0178] Figure 5: Gravimetric vapour sorption isotherms (adsorption and desorption) of Form

[0179] 1 of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2- oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Example 2).

[0180] Figure 6: X-ray powder diffraction pattern (top) of N-[(3-fluoro-4-methoxypyridin-2- yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide following slurry of Form 1 with 90:10 I PA: water. The bottom X-ray powder diffraction pattern is of Form 1 as a reference (Example 2).

[0181] Figure 7: X-ray powder diffraction pattern of Form 2 of N-[(3-fluoro-4-methoxypyridin-2- yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Example 5).

[0182] Figure 8: X-ray powder diffraction pattern of Form 3 of N-[(3-fluoro-4-methoxypyridin-2- yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Example 6).

[0183] Figure 9: X-ray powder diffraction pattern of Form 4 of N-[(3-fluoro-4-methoxypyridin-2- yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Example 7).

[0184] Figure 10: X-ray powder diffraction patterns of Form 1 of N-[(3-fluoro-4-methoxypyridin-

[0185] 2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide during a 25 °C / 60%RH stability study at 0 days (top), 1 month (middle) and 3 months (bottom) (Example 4).

[0186] Figure 11 : X-ray powder diffraction patterns of Form 1 of N-[(3-fluoro-4-methoxypyridin-

[0187] 2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide during a 40 °C / 75%RH stability study at 0 days (top), 1 month (middle) and 3 months (bottom) (Example 4). Figure 12: X-ray powder diffraction pattern of Form 19 of N-[(3-fluoro-4-methoxypyridin-

[0188] 2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Example 9).

[0189] Figure 13: DSC of Form 19 of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-

[0190] (methoxymethyl)-1-({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4- carboxamide (Example 9).

[0191] Figure 14: DSC of Form 1 of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-

[0192] (methoxymethyl)-1-({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4- carboxamide with 1 % of Form 19 of N-[(3-fluoro-4-methoxypyridin-2- yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide added (Example 9).

[0193] Figure 15a: Assay results showing plasma kallikrein inhibition activity of the compound of Formula A and a C1 inhibitor C1-INH in dextran sulfate (DXS)-activated diluted plasma (Example 12).

[0194] Figure 15b: Assay results showing plasma kallikrein inhibition activity of the compound of Formula A and a C1 inhibitor (C1-INH) in DXS-activated undiluted plasma (Example 12).

[0195] Figure 16a: Assay results comparing the plasma kallikrein inhibition activity of the compound of Formula A and C1-INH in DXS-activated diluted plasma (Example 12).

[0196] Figure 16b: Assay results comparing inhibition activity of the compound of Formula A and C1-INH following addition to pre-activated undiluted human plasma. Data are expressed as total fluorescence over time (Fluorescence Units) mean ± SEM of n=3 experiments (Example 12).

[0197] Figure 17a: Assay (bioanalytical) results showing plasma concentrations of the compound of Formula A between 0 and 24 hours post dose, in fasted subjects from eight (8) single ascending dose cohorts (Example 14).

[0198] Figure 17b: Table of Cmax values determined from the assay (bioanalytical) results shown in Figure 17a (Example 14).

[0199] Figure 18a: Assay results showing plasma kallikrein activity in DXS activated undiluted plasma for cohorts 6 to 8 (160 mg, 300 mg, and 600 mg) (Example 14). Figure 18b: Assay results showing the mean plasma kallikrein activity and mean plasma concentration of the compound of Formula A in undiluted plasma in subjects from cohort 8 (600 mg dose) (Example 14).

[0200] Figure 19a: Assay results showing the mean fluorescent kinetic measurements indicating a lag time in catalytic activity during contact system activation in DXS- activated undiluted plasma of a subject who has received a 600 mg dose of the compound of Formula A (Example 14).

[0201] Figure 19b: An enlargement of Figure 19a between 0 and 5 mins following catalytic activation (Example 14).

[0202] Figure 20: Assay results showing mean percent HK protection at selected time points post dosage in DXS activated undiluted plasma for cohorts 6 to 8 (160 mg, 300 mg, and 600 mg), and a representation WES gel image of the immunoblot data (Example 14).

[0203] Figure 21 : Assay results showing the effect of the compound of Formula A on DXS activated HK cleavage at selected time points post dosage in cohort 8 (600mg), and a representation WES gel image of the immunoblot data (Example 14).

[0204] Figure 22: Assay results showing the effect of the compound of Formula A on DXS activated plasma prekallikrein (PPK) cleavage, at selected time points postdosage in cohort 8 (600mg), and a representation WES gel image of the immunoblot data (Example 14).

[0205] Figure 23: Assay results showing the effect of the compound of Formula A on DXS activated generation of FXIIa, at selected time points post-dosage in cohort 8 (600mg), and a representation WES gel image of the immunoblot data (Example 14).

[0206] Figure 24: Assay (bioanalytical) results showing the effect of the plasma concentration of the compound of Formula A at various stages post dose in cohort 8 (600mg) at time points selected for HK, FXIIa, PPK analysis (Example 14).

[0207] Figure 25: Assay results showing no significant food effect on the plasma kallikrein inhibitory activity of the compound of Formula A in DXS-activated undiluted plasma (Example 14). Figures 26a, b: Assay results showing a time course of dextran sulfate activated cleavage of HK in HAE whole undiluted plasma determined using western blotting, and a representative blot image (Example 15).

[0208] Figures 27a, b: Assay results showing the dose response of the compound of Formula A on full length HK levels in dextran sulfate activated healthy control plasma and HAE plasma, and representative WES system gel images (Example 15).

[0209] Figure 28a: Preliminary pharmacokinetic data from the phase 2 study (Example 16).

[0210] Figure 28b: PK profile of HAE patients from part 1 of the phase 2 study, overlaid with PK data from healthy volunteers (Example 16).

[0211] Figure 28c: Graph showing cumulative % of patients that used rescue medication within 12 hours of being administered the compound (Example 16).

[0212] Figure 28d: Graph showing cumulative % of patients with symptom relief (as assessed using the 7TQ method) within 12 hours of being administered the compound (Example 16).

[0213] Figure 28e: Graph showing comparison of time to symptom relief data for the compound with known clinical data for approved injectable product Ruconest® (Example 16).

[0214] Figure 28f: Graph showing cumulative % of patients with symptom relief (as assessed using the composite VAS score) within 12 hours of being administered the compound (Example 16).

[0215] Figure 28g: Graph showing mean composite VAS score within 12 hours of being administered the compound (Example 16).

[0216] Figure 28h: Graph showing comparison of mean composite VAS score for patients treated with the compound with known clinical data for approved injectable product Firazyr® (active ingredient, icatibant) (Example 16).

[0217] Figure 28i: Graph showing the % of HAE attacks that did not worsen (as assessed using the 5LS method) within 12 hours of being administered the compound (Example 16).

[0218] Figure 29a: Mean plasma concentrations over time of 4 cohorts in the phase 1 multiple dose study (Example 17). Figure 29b: Mean plasma concentrations over time (semi-logarithmic scale) of 4 cohorts in the phase 1 multiple dose study (Example 17).

[0219] Figure 30: X-ray powder diffraction pattern of Form 5 (freebase) of N-[(3-fluoro-4- methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Example 8B).

[0220] Figure 31 : DSC thermograph of Form 5 (freebase) of N-[(3-fluoro-4-methoxypyridin-2- yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Example 8B).

[0221] Figure 32: Infrared (IR) spectrum of Form 19 of N-[(3-fluoro-4-methoxypyridin-2- yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Example 9).

[0222] Figure 33: DSC thermograph of Form 1 (Example 9A).

[0223] Figure 34: DSC thermograph of Form 19 (Example 9A).

[0224] Figure 35: DSC thermograph of a sample of 99.5 wt.% Form 1 and 0.5 wt.% Form 19

[0225] (Example 9A).

[0226] Figure 36: Overlaid DSC thermographs of Form 1 with 0.1 wt.%, 0.2 wt.%, 0.5 wt.%, 1.0 wt.% or 2.0 wt.% of Form 19 (Example 9A).

[0227] Figure 37: Flow diagram for steps of the screening method (Example 9A).

[0228] Embodiments provided herein may be more fully understood by reference to the following examples. These examples are meant to be illustrative of treatments provided herein but are not in any way limiting. Indeed, the scope of the invention is defined by the claims.

[0229] While examples of certain embodiments are provided herein, it will be apparent to those skilled in the art that various changes and modifications may be made. Such modifications are also intended to fall within the scope of the appended claims.

[0230] General Experimental Details

[0231] In the following examples, the following abbreviations and definitions are used:

[0232] Instruments and Methods

[0233] / . General

[0234] All reactions were carried out under an atmosphere of nitrogen unless specified otherwise.

[0235] All solvents and commercial reagents were used as received. Chemical names were generated using automated software such as the Autonom software provided as part of the ISIS Draw package from MDL Information Systems or the Chemaxon software provided as a component of MarvinSketch or as a component of the I DBS E- WorkBook.

[0236] / / . Nuclear Magnetic Resonance (NMR)

[0237] 1H NMR spectra were recorded on a Bruker (400MHz) or on a JEOL (400MHz) spectrometer with reference to deuterium solvent and at rt.

[0238] / / / . Mass Spectrometry (MS)

[0239] Molecular ions were obtained using LCMS which was carried out using a Chromolith Speedrod RP-18e column, 50 x 4.6 mm, with a linear gradient 10% to 90% 0.1% HCO2H / MeCN into 0.1% HCO2H / H2O over 13 min, flow rate 1.5 mL / min, or using Agilent, X-Select, acidic, 5-95% MeCN / water over 4 min. Data was collected using a Thermofinnigan Surveyor MSQ mass spectrometer with electospray ionisation in conjunction with a Thermofinnigan Surveyor LC system.

[0240] Alternatively, molecular ions were obtained using LCMS which was carried out using an Agilent Poroshell 120 EC-C18 (2.7 m, 3.0 x 50mm) column with 0.1% v / v Formic acid in water [eluent A]; Macon [eluent B]; Flow rate 0.8ml_ / min and 1.5 minutes equilibration time between samples, gradient shown below. Mass detection was afforded with API 2000 mass spectrometer (electrospray).

[0241] Gradient:

[0242] Table 1

[0243] IV. Flash Chromatography Where products were purified by flash chromatography, ‘silica’ refers to silica gel for chromatography, 0.035 to 0.070 mm (220 to 440 mesh) (e.g. Merck silica gel 60), and an applied pressure of nitrogen up to 10 p.s.i accelerated column elution. Reverse phase preparative HPLC purifications were carried out using a Waters 2525 binary gradient pumping system at flow rates of typically 20 mL / min using a Waters 2996 photodiode array detector.

[0244] V. X-Ray Powder Diffraction

[0245] Method A

[0246] X-Ray Powder Diffraction patterns were collected on a Philips X-Pert MPD diffractometer and analysed using the following experimental conditions (Method A), unless otherwise specified: Tube anode: Cu

[0247] Generator tension: 40 kV

[0248] Tube current: 40 mA

[0249] Wavelength alphal : 1 .5406 A

[0250] Wavelength alpha2: 1.5444 A

[0251] Start angle

[0020] : 4

[0252] End angle

[0020] : 40

[0253] Continuous scan

[0254] Approximately 2 mg of sample under analysis was gently compressed on the XRPD zero back ground single obliquely cut silica sample holder. The sample was then loaded into the diffractometer for analysis.

[0255] Method B

[0256] Where specified, X-Ray Powder Diffraction patterns were collected using the following method (Method B):

[0257] X-ray powder diffraction studies were performed using a Bruker AXS D2 PHASER (D2- 205355) in Bragg- Brentano configuration, equipment #2353. A Cu anode at 30kV, 10 mA, sample stage standard rotating (5 / min) with beam stop and monochromatisation by a Kp-filter (0.59% Ni) are used. The slits that are used are fixed divergence slits 1.0mm (=0.61°), primary axial Soller slit 2.5° and secondary axial Soller slit 2.5°. The detector is a linear detector LYNXEYE with receiving slit 5° detector opening. The standard sample holder (0.1mm cavity in (510) silicon wafer) has a minimal contribution to the background signal. The measurement conditions: scan range 5 — 45° 20, sample rotation 5 rpm,

[0258] 0.5s / step, 0.0107step, 3.0mm detector slit; and all measuring conditions are logged in the instrument control file. The software used for data collection is Diffrac.Commander v4.0. Data analysis is performed using Diffrac.Eva V4.1 evaluation software. No background correction or smoothing is applied to the patterns.

[0259] Method C

[0260] Where specified, X-Ray Powder Diffraction patterns were collected using a Diffractometer X- ray D8 ADVANCE with PSD detector (model 1-D Si first generation “Lynxeye” Brucker) with the following experimental conditions:

[0261] Table 2

[0262] VI. Differential Scanning Calorimetry (DSC)

[0263] Method A

[0264] DSC data were collected using the following method: Approximately 5 mg of each sample was weighed into an aluminium DSC pan and sealed non-hermetically with an aluminium lid. The sample was then loaded into a Perkin-Elmer Jade DSC and held at 30°C. Once a stable heatflow response was obtained, the sample was then heated to a temperature between 200 and 300°C at a scan rate of 10°C / min and the resulting heat flow response was monitored. A 20 cm3 / min helium purge was used. Prior to analysis, the instrument was temperature and heat flow verified using an indium standard. Method B

[0265] Where specified, DSC data were collected using a Q100 V9.9 Build 303 DSC with approximately 2-3 mg of sample in an aluminium pan. The aluminium pan is sealed and the sample purge flow under nitrogen is at 50 mL / min. The sample was heated to a temperature of 300°C at a scan rate of 10°C / min and the resulting heat flow response was monitored.

[0266] Method C

[0267] Where specified, DSC data were collected using the following method: approximately 5 mg of a sample was weighed into a standard aluminium crucible (40pL) - Mettler 27331 and sealed hermetically with an aluminium lid. The sample was then loaded into a Mettler Toledo DSC1 system. The sample was heated from 50°C to 250°C at 10 °C / min. A 200 mL / min nitrogen purge was used. The software used for instrument control and data analysis was STARe v12.10.

[0268] VII. Simultaneous Thermal Analysis (STA)

[0269] Simultaneous Thermal Analysis (STA) data were collected using the following method: Approximately 5 mg of sample was accurately weighed into a ceramic crucible and it was placed into the chamber of Perkin-Elmer STA 600 TGA / DTA analyser at ambient temperature. The sample was then heated at a rate of 10°C / min, typically from 25°C to 300°C, during which time the change in weight was monitored as well as DTA signal. The purge gas used was nitrogen at a flow rate of 20 cm3 / min.

[0270] VIII. Infrared (IR) Spectroscopy

[0271] Infrared spectroscopy data were collected using a Perkin Elmer FTIR equipped with a diamond ATR probe. Samples was scanned from 4000 to 400 cm-1.

[0272] Synthetic Methods

[0273] Example 1 - N-[(3-Fluoro-4-methoxypyridin-2-yl)methyll-3-(methoxymethyl)-1-( / 4-[(2- oxopyridin- 1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form A)

[0274] A. 1-(4-Hydroxymethyl-benzyl)-1H-pyridin-2-one 4-(Chloromethyl)benzylalcohol (5.0 g, 31.93 mmol) was dissolved in acetone (150 mL). 2- hydroxypyridine (3.64 g, 38.3 mmol) and potassium carbonate (13.24 g, 95.78 mmol) were added and the reaction mixture was stirred at 50 °C for 3 hrs after which time the solvent was removed in vacuo and the residue taken up in chloroform (100 mL). This solution was washed with water (30 mL), brine (30 mL), dried (Na2SO4) and evaporated in vacuo. The residue was purified by flash chromatography (silica), eluent 3% MeOH 1 97% CHCh, to give a white solid identified as 1-(4-hydroxymethyl-benzyl)-1 H-pyridin-2-one (5.30g, 24.62mmol, 77% yield). [M+Na]+= 238

[0275] B. 1-(4-Chloromethyl-benzyl)-1H-pyridin-2-one

[0276] 1-(4-Hydroxymethyl-benzyl)-1 H-pyridin-2-one (8.45 g, 39.3 mmol), dry DCM (80 mL) and triethylamine (7.66 ml, 55.0 mmol) were cooled in an ice bath. Methanesulfonyl chloride (3.95 ml, 51.0 mmol) was added and stirred in ice bath for 15 min. The ice bath was removed and stirring continued at rt temperature overnight. The reaction mixture was partitioned between DCM (100 mL) and saturated aqueous NH4CI solution (100 mL). The aqueous layer was extracted with further DCM (2 x 50 mL) and the combined organics washed with brine (50 mL), dried over Na2SO4, filtered and concentrated to give 1-(4-chloromethyl-benzyl)-1 H-pyridin-2- one (8.65 g, 36.6 mmol, 93 % yield) as a pale yellow solid.

[0277] [MH]+= 234.1

[0278] C. Methyl 3-(methoxymethyl)-1-(4-((2-oxopyridin-1(2H)-yl)methyl)benzyl)-1 H-pyrazole- 4-carboxylate

[0279] Potassium carbonate (519 mg, 3.76 mmol) was added to a solution of methyl 3- (methoxymethyl)-1 H-pyrazole-4-carboxylate (320 mg, 1.88 mmol; CAS no. 318496-66-1 (synthesised according to the method described in WO 2012 / 009009)) and 1-(4- (chloromethyl)benzyl)pyridin-2(1 H)-one (527 mg, 2.26 mmol) in DMF (5 mL) and heated at 60 °C overnight. The reaction mixture was diluted with EtOAc (50 mL) and washed with brine (2 x 100 mL), dried over magnesium sulfate, filtered and reduced in vacuo. The crude product was purified by flash chromatography (40 g column, 0-100% EtOAc in isohexanes) to afford two regioisomers. The second isomer off the column was collected to afford methyl 3- (methoxymethyl)-1-(4-((2-oxopyridin-1(2H)-yl)methyl)benzyl)-1 H-pyrazole-4-carboxylate (378 mg, 1.01 mmol, 53.7 % yield) as a colourless gum.

[0280] [MH]+= 368.2

[0281] D. 3-(Methoxymethyl)-1-(4-((2-oxopyridin-1(2H)-yl)methyl)benzyl)-1H-pyrazole-4- carboxylic acid

[0282] To methyl 3-(methoxymethyl)-1-(4-((2-oxopyridin-1(2H)-yl)methyl)benzyl)-1 H-pyrazole-4- carboxylate (3.77 g, 10.26 mmol) in THF (5 mL) and MeOH (5 mL) was added 2M NaOH solution (15.39 ml, 30.8 mmol) and stirred at rt overnight. 1M HCI (50 mL) was added and extracted with EtOAc (50 mL). The organic layer was washed with brine (50 mL), dried over magnesium sulfate, filtered and reduced in vacuo to give 3-(methoxymethyl)-1-(4-((2- oxopyridin-1 (2H)-yl)methyl)benzyl)-1 H-pyrazole-4-carboxylic acid (1.22 g, 3.45 mmol, 33.6 % yield) as a white powder.

[0283] [MH]+= 354.2

[0284] E. 3-Fluoro-4-methoxy-pyridine-2-carbonitrile

[0285] To a large microwave vial, copper (I) cyanide (1.304 g, 14.56 mmol) was added to a solution of 2-bromo-3-fluoro-4-methoxypyridine (1 g, 4.85 mmol) in DMF (5 mL). The reaction vial was sealed and heated to 100 °C for 16 hrs. The reaction mixture was diluted with water (20 mL) and EtOAc (20 mL). The thick suspension was sonicated and required additional water (40 mL) and EtOAc (2 x 50 mL) with sonication to break-up the solid precipitated. The combined layers were filtered through a plug of celite and the organic layer isolated, washed with brine (50 mL), dried over magnesium sulfate, filtered and the solvent removed under reduced pressure to give a pale green solid identified as the desired compound 3-fluoro-4-methoxy- pyridine-2-carbonitrile (100 mg, 0.578 mmol, 12 % yield)

[0286] F. (3-Fluoro-4-methoxy-pyridin-2-ylmethyl)-carbamic acid tert-butyl ester

[0287] 3-Fluoro-4-methoxy-pyridine-2-carbonitrile (100 mg, 0.578 mmol) was dissolved in anhydrous methanol (10 mL, 247 mmol) and nickel chloride hexahydrate (14 mg, 0.058 mmol) was added followed by di-tert-butyl dicarbonate (255 mg, 1.157 mmol). The resulting pale green solution was cooled in an ice-salt bath to -5 °C and then sodium borohydride (153 mg, 4.05 mmol) was added portionwise maintaining the reaction temperature ~0 °C. The deep brown solution was left to stir at 0 °C and slowly allowed to warm to rt and then left to stir at rt for 3 hrs. The reaction mixture was evaporated to dryness at 40 °C to afford a black residue which was diluted with DCM (10 mL) and washed with sodium hydrogen carbonate (10 mL). An emulsion formed so the organics were separated via a phase separating cartridge and concentrated. The crude liquid was purified by chromatography eluting with EtOAc / iso-Hexane to afford the title compound, (3-fluoro-4-methoxy-pyridin-2-ylmethyl)-carbamic acid tert-butyl ester as a clear yellow oil (108 mg, 62 % yield)

[0288] [MH]+= 257

[0289] G. C-(3-Fluoro-4-methoxy-pyridin-2-yl)-methylamine hydrochloride salt

[0290] (3-Fluoro-4-methoxy-pyridin-2-ylmethyl)-carbamic acid tert-butyl ester (108mg, 0.358mmol) was taken up in iso-propyl alcohol (1 mL) and then HCI (6N in iso-propyl alcohol) (1 mL, 0.578 mmol) was added at rt and left to stir at 40 °C for 2 hrs. The reaction mixture was concentrated under reduced pressure and then triturated with ether, sonicated and then decanted to give a cream coloured solid (75 mg, 55% yield) identified as C-(3-fluoro-4-methoxy-pyridin-2-yl)- methylamine hydrochloride salt.

[0291] [MH]+= 157

[0292] H. N-[(3-Fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin- 1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form A)

[0293] 3-(Methoxymethyl)-1-(4-((2-oxopyridin-1(2H)-yl)methyl)benzyl)-1 H-pyrazole-4-carboxylic acid (825 mg, 2.34 mmol) and C-(3-fluoro-4-methoxy-pyridin-2-yl)-methylamine hydrochloride salt (450 mg, 2.34 mmol) were dissolved in DCM while cooling to 0°C. 1-Ethyl-3-(3- dimethylaminopropyl)carbodiimide hydrochloride (627.0 mg, 3.27 mmol), HOBt (378.8 mg, 2.80 mmol) and triethylamine (1.63 mL, 1182 mmol) were added while stirring, the mixture allowed to warm to rt and stirring continued for 20 hrs. Chloroform (50 mL) was added, the mixture was washed with saturated NaHCOs(aq) and reduced in vacuo. The crude material was purified by chromatography eluting with methanol / DCM. The resulting solid was dissolved in hot MeCN, allowed to cool and precipitate, and the resulting solids were removed by filtration. The filtrate was reduced in vacuo then freeze dried from MeCN / water to afford the title compound as a white solid (720 mg, 62% yield).

[0294] [MH]+= 492.0

[0295] NMR (CD3OD) 5: 3.41 (3H, s), 4.03 (3H, s), 4.65 (2H, s), 4.72 (2H, d, J=2.3Hz), 5.24 (2H, s),

[0296] 5.37 (2H, s), 6.44 (1 H, td, J = 1.4, 6.8Hz), 6.62 (1 H, d, J = 9.0Hz), 7.18-7.22 (1 H, m), 7.31-

[0297] 7.38 (4H, m), 7.56-7.60 (1 H, m), 7.75 (1 H, dd, J = 1.9, 7.1 Hz), 8.18 (1 H, s), 8.27 (1 H, d, J = 5.6Hz) ppm.

[0298] An XRPD diffractogram of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1- ({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form A) is shown in Figure 1. Form A was found to be mostly amorphous.

[0299] Example 2 - N-[(3-Fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-( 4-[(2- oxopyridin- 1-yl)methyllphenyl}methyl)pyrazole-4-carboxamide (Form 1)

[0300] 3-(Methoxymethyl)-1-(4-((2-oxopyridin-1(2H)-yl)methyl)benzyl)-1 H-pyrazole-4-carboxylic acid (825 mg, 2.34 mmol) and C-(3-fluoro-4-methoxy-pyridin-2-yl)-methylamine hydrochloride salt (450 mg, 2.34 mmol) were dissolved in DCM while cooling to 0°C. 1-Ethyl-3-(3- dimethylaminopropyl)carbodiimide hydrochloride (627.0 mg, 3.27 mmol), HOBt (378.8 mg, 2.80 mmol) and triethylamine (1.63 mL, 1182 mmol) were added while stirring, the mixture allowed to warm to rt and stirring continued for 20 hrs. Chloroform (50 mL) was added, the mixture was washed with saturated NaHCOs(aq) and reduced in vacuo. The crude material was purified by chromatography eluting with methanol / DCM. The solvent was removed in vacuo and the resulting solid triturated with diethyl ether. The resulting solids were collected by filtration to afford the title compound.

[0301] An XRPD diffractogram of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1- ({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 1) is shown in Figure 2a.

[0302] Peak position table:

[0303] Table 3

[0304] The STA data for Form 1 are shown in Figure 3.

[0305] The DSC data for Form 1 are shown in Figure 4.

[0306] The GVS data for Form 1 are listed in the table below and shown in Figure 5.

[0307] Table 4

[0308] Slurry study Form 1 (20 mg) was suspended in 90 / 10 IPA / water (200pL or 300pL) and shaken at ambient temperature for 72 hrs. The supernatant was evaporated rather than filtered due to the small volume and the resulting solid was examined by XRPD (Figure 6). The resulting XRPD (Figure 6) was different to that of Figure 2a which indicated that the free base probably has a tendency to form hydrate(s).

[0309] Visual aqueous solubility

[0310] Form 1 (10mg) was weighed into a glass vial and water was added in 100pL portions up to 3mL then 1mL portions thereafter. Solubility was assessed visually following a brief period of equilibration.

[0311] Form 1 did not give any indication it was dissolving at all in 20mL water (« 0.5mg / mL).

[0312] Example 3 - N-[(3-Fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-( / 4-[(2- oxopyridin- 1-yl)methyllphenyl}methyl)pyrazole-4-carboxamide (Form 1)

[0313] 3-(Methoxymethyl)-1-(4-((2-oxopyridin-1(2H)-yl)methyl)benzyl)-1 H-pyrazole-4-carboxylic acid (825 mg, 2.34 mmol) and C-(3-fluoro-4-methoxy-pyridin-2-yl)-methylamine hydrochloride salt (450 mg, 2.34 mmol) were dissolved in DCM while cooling to 0°C. 1-Ethyl-3-(3- dimethylaminopropyl)carbodiimide hydrochloride (627.0 mg, 3.27 mmol), HOBt (378.8 mg, 2.80 mmol) and triethylamine (1.63 mL, 1182 mmol) were added while stirring, the mixture allowed to warm to rt and stirring continued for 20 hrs. Chloroform (50 mL) was added, the mixture was washed with saturated NaHCOs(aq) and reduced in vacuo. The crude material was purified by chromatography eluting with methanol / DCM. The resulting solid was dissolved in hot MeCN, allowed to cool and precipitate, and the resulting solids were collected by filtration to afford the title compound as a white solid (130 mg, 11% yield).

[0314] An XRPD diffractogram (recorded using Method B) of N-[(3-fluoro-4-methoxypyridin-2- yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4- carboxamide (Form 1) is shown in Figure 2b. The XRPD diffractogram (Figure 2b) of the isolated solids confirmed that they were of the same polymorphic form as Form 1 (Example 2) (Figure 2a).

[0315] Peak position table: Table 5

[0316] Example 4 - N-[(3-Fluoro-4-methoxypyridin-2-yl)methyll-3-(methoxymethyl)-1-( / 4-[(2- oxopyridin- 1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 1) 3-(Methoxymethyl)-1-(4-((2-oxopyridin-1(2H)-yl)methyl)benzyl)-1 H-pyrazole-4-carboxylic acid (61 g, 0.173 mol) was dissolved in DMF (400 mL) and 1 ,1 ’-carbonyldiimidazole (27.99 g, 0.173 mol) was added portion wise. Once the addition was complete, the reaction was heated to 50 °C for 2 hrs. C-(3-fluoro-4-methoxy-pyridin-2-yl)-methylamine (26.95 g, 0.17 3mol) was added to the reaction mixture portion wise. The reaction was heated to 50 °C overnight. The reaction was cooled to rt and added dropwise to a 3:1 mixture of water and saturated NaHCOs(aq) (4000 mL). The resulting suspension was stirred for 30 min before isolating the solids by filtration. The solids were washed with water (2x500 mL) before drying in a vacuum oven to give 119 g of the crude product. The crude product was combined with two other separate batches (starting with 0.173 mol and 0.0874 mol of the acid starting material respectively) and slurried together in IPA (1400 mL) and heated to reflux. Additional portions of IPA were added until all of the material had dissolved at reflux (total of 2000 mL IPA added). The solution was held at reflux for 30 min before it was cooled to rt. The mixture was cooled further with an ice / water bath for 30 min before the product was collected by filtration. The solids were washed with IPA and dried to give 167.2 g of the title product (78.5% yield).

[0317] [MH]+= 491.9

[0318] NMR (CD3OD) spectrum conformed to the NMR spectrum of Example 1.

[0319] An XRPD diffractogram (recorded using Method B) of the isolated solids (Figure 2c) confirmed that they were of the same polymorphic form as Form 1 (Example 2 and Example 3) (Figures 2a and 2b).

[0320] Stability data

[0321] A sample of Form 1 was packed in double polyethylene bags and sealed in a HDPE bottle and stored at conditions of 25 °C 160%RH. The sample was reanalysed after 1 month and 3 months by XRPD (using Method B). The data is shown in Figure 10. No change in the XRPD diffractogram was observed when the sample was stored at 25 °C I 60%RH after either 1 month or 3 months.

[0322] Further tests on the sample of Form 1 stored at 25 °C 160%RH were carried out as described in the table below:

[0323] Table 6

[0324]

[0325] A second sample of Form 1 was packed in double polyethylene bags and sealed in a HDPE bottle and stored under accelerated stability conditions of 40 °C I 75%RH. The sample was reanalysed after 1 month and 3 months by XRPD (using Method B). The data is shown in Figure 11. No change in the XRPD diffractogram was observed when the sample was stored at 40 °C 1 75%RH after either 1 month or 3 months.

[0326] Further tests on the sample of Form 1 stored at 40 °C 175%RH were carried out as described in the table below:

[0327] Table 7

[0328]

[0329] Example 5 - N-[(3-Fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-( / 4-[(2- oxopyridin- 1-yl)methyllphenyl}methyl)pyrazole-4-carboxamide (Form 2)

[0330] N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (19.5 mg) in 90 / 10 IPA / water (100 pL) was heated to dissolve the solid, filtering if necessary. The resulting solution was cooled by plunging the warm solution into a liquid nitrogen bath. The sample was then transferred into a freezer. The resulting solids were isolated to afford N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]- 3-(methoxymethyl)-1-({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 2).

[0331] An XRPD diffractogram of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1- ({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 2) is shown in Figure 7.

[0332] Peak position table:

[0333] Table 8

[0334] Example 6 - N-[(3-Fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-( / 4-[(2- oxopyridin- 1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 3)

[0335] A suspension of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2- oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (30 mg) in 50 / 50 methanol / water (100 pL) was matured by temperature cycling for 2 days. The resulting solids were isolated to afford N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4- [(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 3).

[0336] An XRPD diffractogram of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1- ({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 3) is shown in Figure 8.

[0337] Peak position table:

[0338] Table 9

[0339] Example 7 - N-[(3-Fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-( / 4-[(2- oxopyridin- 1-yl)methyllphenyl}methyl)pyrazole-4-carboxamide (Form 4)

[0340] A suspension of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2- oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (20 mg) in 50 / 50 methanol / water (100 pL) was heated. Not all of the solid dissolved and therefore the mixture was filtered. The filtrate was allowed to evaporate under nitrogen to afford N-[(3-fluoro-4- methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 4).

[0341] An XRPD diffractogram of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1- ({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 4) is shown in Figure 9.

[0342] Example 8A - N-[(3-Fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-(f4-[(2- oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 5 (freebase))

[0343] A suspension of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2- oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (500 mg) in 50 / 50 methanol / water (2.5 mL) was matured by temperature cycling from 20°C to 50°C for 24 hours. The resulting solids were isolated to afford N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3- (methoxymethyl)-1-({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 5).

[0344] Example 8B- N-[(3-Fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)- 1-( / 4-[ (2- oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 5 (freebase))

[0345] A solution of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2- oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (500 mg) in 50 / 50 methanol / water (20 mL) was prepared. The solution was allowed to evaporate to dryness under vacuum to afford N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4- [(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 5).

[0346] An XRPD diffractogram of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1- ({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 5) obtained from Example 8B is shown in Figure 30. This XRPD diffractogram was obtained using Method B above.

[0347] Peak position table of most prominent peaks:

[0348] Table 10

[0349] Form 5 is less stable than Forms 1, 2 or 3 and it converts into polymorph Form 3, over time. Performing an XRPD measurement on Form 5 resulted in Form 3.

[0350] Differential Scanning Calorimetry (DSC)

[0351] TGA / DSC studies were performed using a Mettler Toledo TGA / DSC1 Stare System, equipment #1547, auto-sampler equipped, using pin-holed Al-crucibles of 40 pl. Measurement conditions: 5 min 30.0 °C, 30.0 - 350.0 °C with 10 °C / min., N2 flow of 40 ml / min. The software used for instrument control and data analysis is STARe v12.10. The TGA / DSC data for Form 5 are shown in Figure 31.

[0352] Example 9 - N-[(3-Fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-( / 4-[(2- oxopyridin- 1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide (Form 19)

[0353] Into a reactor was charged 5g of Form 1 of the compound of Formula A then 40 ml of isopropanol. The suspension was heated to reflux (82°C). The majority of the suspended solid gradually went into solution, however the mixture remained very cloudy, with the formation of a product crust on the reactor wall which did not dissolve. The mixture was maintained at 78-80°C for 1h. During the hold at this temperature, the mixture crystallised notably to give an opaque suspension. The suspension was cooled to 0-5°C over 1 h, then the solid was filtered off, washed with 20ml of isopropanol and dried under vacuum at 35- 40°C. Analysis by differential scanning and XRPD of the solid obtained confirmed that Form 19 had been produced.

[0354] An XRPD diffractogram of Form 19 is shown in Figure 12 (Method C).

[0355] Peak position table:

[0356] Table 11

[0357] An IR spectrum of Form 19 is shown in Figure 32. Peak position table:

[0358] Table 12

[0359] DSC data for Form 19 is shown in Figure 13. Form 19 exhibits an endothermic peak in its DSC thermograph at about 162.59°C.

[0360] A DSC trace was also obtained for a sample of Form 1 of the compound of Formula A to which had been added 1 % of Form 19 of the compound of Formula A. The resulting DSC trace is shown in Figure 14.

[0361] Example 9A - Screening method for detecting the presence of Form 19 in a test sample of Form 1

[0362] A screening method was developed to determine whether less than 0.5 wt.% of Form 19 is present in a sample of essentially pure Form 1. The screening method was developed based on DSC Method C.

[0363] Thermographs obtained for samples of Form 1 and Form 19 using Method C had endotherms at 151.07°C (Tonset) and 158.60°C (Tonset), respectively (Figures 33 and 34). Visual examination of thermographs obtained for mixtures of Form 1 and Form 19 (0.1 wt.%, 0.2 wt.%, 0.5 wt.%, 1.0 wt.% or 2.0 wt.%) confirmed that DSC Method C can detect 0.1 wt.% of Form 19 in Form 1 (Figure 36).

[0364] Thermographs were obtained using DSC Method C with increased (12°C / min) or decreased (8°C / min) heating rates. For a sample of Form 1 , the TonSet of the observed thermograph did not vary significantly from that observed with the normal heating rate (10°C / min). At 8°C / min and 12°C / min, the thermographs of mixtures of Form 1 and 0.1 wt.% Form 19 showed an endotherm for Form 19.

[0365] Procedure

[0366] A reference mixture of 99.5 wt.% of Form 1 and 0.5 wt.% of Form 19 was prepared. Form 1 may be prepared using the method of Example 2 or Example 4. Form 19 may be prepared using the method of Example 9. The mixture was prepared by adding the Form 1 and the Form 19 to a 30 ml glass bottle and homogenising with a 3-dimensional shaker, TurbulaT2F.

[0367] One crucible of the reference mixture was prepared and analysed using DSC Method C. The thermograph obtained was traced and integrated (Figure 35). Suitability of the reference mixture was confirmed by the presence of an endotherm at approximately 151 °C (onset), which corresponds to Form 1 , and an endotherm at approximately 158°C (onset), which corresponds to Form 19.

[0368] Three crucibles of a test sample of Form 1 were prepared and analysed using DSC Method C. The obtained thermographs for the reference mixture and the three test samples were traced and integrated. The presence of Form 19 in the test samples was confirmed by the presence of an endotherm at approximately 158°C (onset).

[0369] If an endotherm at approximately 158°C (onset) was present in the thermographs of the test samples, then for the reference mixture and each text sample, the ratio of melting enthalpies was calculated according to Formula 1 :

[0370] Where: • HfUSForm 19) is the melting enthalpy for the quantity of Form 19 present in the sample obtained by integrating the endotherm at approximately 158°C (onset) in each thermograph.

[0371] • AHfus(Form 1) is the melting enthalpy for the quantity of Form 1 present in the sample obtained by integrating the endotherm at approximately 151 °C (onset) in each thermograph.

[0372] The melting enthalpies were normalised according to the mass of the test sample or reference mixture analysed.

[0373] The ratio calculated for the reference mixture (Rr) was then compared to each of the ratios calculated for the test samples (Rt) to determine whether less than 0.5 wt.% of Form 19 was present in each of the test samples. For each case it was determined which of three cases was true:

[0374] (i) Rt < (Rrx 0.75)

[0375] (ii) (Rrx 0.75) < Rt< Rr; or

[0376] (iii) Rt> Rr

[0377] If case (i) were true, it was determined that the test sample contained less than 0.5 wt.% of Form 19, this was considered a “pass”. If case (ii) were true for at least one of the test samples, then the procedure would be repeated with three new test samples. If case (iii) were true, it was determined that the test sample contained more than 0.5 wt.% of Form 19, this was considered a “fail”.

[0378] A flow diagram for the steps of the screening method is shown in Figure 37.

[0379] Example 10 - Biological Assays

[0380] The ability of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2- oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide to inhibit plasma kallikrein may be determined using the following biological assays:

[0381] Determination of the IC50 for plasma kallikrein

[0382] Plasma kallikrein inhibitory activity in vitro was determined using standard published methods (see e.g. Johansen et al., Int. J. Tiss. Reac. 1986, 8, 185; Shori et al., Biochem. Pharmacol., 1992, 43, 1209; Sturzebecher et al., Biol. Chem. Hoppe-Seyler, 1992, 373, 1025). Human plasma kallikrein (Protogen) was incubated at 25 °C with the fluorogenic substrate H-Dpro- Phe-Arg-AFC and various concentrations of the test compound. Residual enzyme activity (initial rate of reaction) was determined by measuring the change in optical absorbance at 410nm and the IC50 value for the test compound was determined.

[0383] When tested in this assay, N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1- ({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide showed an IC50 (human Pkal) of 3.3 nM.

[0384] N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide was also screened for inhibitory activity against the related enzyme KLK1 using the following biological assay:

[0385] Determination of the IC50 for KLK1

[0386] KLK1 inhibitory activity in vitro was determined using standard published methods (see e.g. Johansen et al., Int. J. Tiss. Reac. 1986, 8, 185; Shori et al., Biochem. Pharmacol., 1992, 43, 1209; Sturzebecher et al., Biol. Chem. Hoppe-Seyler, 1992, 373, 1025). Human KLK1 (Callbiochem) was incubated at 25 °C with the fluorogenic substrate H-Dval-Leu-Arg-AFC and various concentrations of the test compound. Residual enzyme activity (initial rate of reaction) was determined by measuring the change in optical absorbance at 410nm and the IC50 value for the test compound was determined.

[0387] When tested in this assay, N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1- ({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide showed an IC50 (human KLK1) of >40000 nM.

[0388] N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1-({4-[(2-oxopyridin-1- yl)methyl]phenyl}methyl)pyrazole-4-carboxamide was also screened for inhibitory activity against the related enzyme FXIa using the following biological assay:

[0389] Determination of the % inhibition for FXIa

[0390] FXIa inhibitory activity in vitro was determined using standard published methods (see e.g. Johansen et al., Int. J. Tiss. Reac. 1986, 8, 185; Shori et al., Biochem. Pharmacol., 1992, 43, 1209; Sturzebecher et al., Biol. Chem. Hoppe-Seyler, 1992, 373, 1025). Human FXIa (Enzyme Research Laboratories) was incubated at 25 °C with the fluorogenic substrate Z-Gly- Pro-Arg-AFC and 40 pM of the test compound. Residual enzyme activity (initial rate of reaction) was determined by measuring the change in optical absorbance at 410nm. When tested in this assay, N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1- ({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide showed a % inhibition @ 40 pM (human FXIa) of 0%.

[0391] Example 11 - Pharmacokinetic Study

[0392] A pharmacokinetic study of N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3-(methoxymethyl)-1- ({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide was performed to assess the pharmacokinetics following a single oral dose in male Sprague-Dawley rats. Two rats were given a single po dose of 5 mL / kg of a nominal 2 mg / mL (10 mg / kg) composition of test compound in vehicle. Following dosing, blood samples were collected over a period of 24 hrs. Sample times were 5, 15 and 30 minutes then 1 , 2, 4, 6, 8 and 12 hrs. Following collection, blood samples were centrifuged and the plasma fraction analysed for concentration of test compound by LCMS.

[0393] Oral exposure data acquired from this study for N-[(3-fluoro-4-methoxypyridin-2-yl)methyl]-3- (methoxymethyl)-1-({4-[(2-oxopyridin-1-yl)methyl]phenyl}methyl)pyrazole-4-carboxamide is shown below:

[0394] Table 13

[0395] Example 12 - Comparison of the compound of Formula A with a C1 inhibitor (C1-INH)

[0396] Aim: To identify the biochemical and biophysical properties of the compound of Formula A that contribute to its optimal efficacy in controlling the Kallikrein Kinin System in plasma. These properties are then compared to C1-INH as a therapeutic benchmark for HAE.

[0397] Methods: Plasma kallikrein inhibitory activity in vitro was determined using standard published methods (see e.g. Johansen et al., Int. J. Tiss. Reac. 1986, 8, 185; Shori et al., Biochem. Pharmacol., 1992, 43, 1209; Sturzebecher et al., Biol. Chem. Hoppe-Seyler, 1992, 373, 1025). Human plasma kallikrein (Protogen) was incubated at 25 °C with the fluorogenic substrate H-Dpro- Phe-Arg-AFC and various concentrations of the test compound. Residual enzyme activity (initial rate of reaction) was determined by measuring the change in optical absorbance at 410nm and the IC50 value for the test compound was determined.

[0398] The rate of formation of the enzyme-inhibitor complex (Kon) was determined using purified Pka rapidly mixed with a solution containing fluorogenic substrate and a concentration range of inhibitor. The time-dependent establishment of inhibition was then used to calculate the rate of formation of the enzyme-inhibitor complex for each concentration of inhibitor. The Konwas calculated by plotting the rate of inhibition versus the inhibitor concentration. Data in Table 14 are presented in pM-1sec1.

[0399] Catalytic activity of Pka in dextran sulfate-activated (DXS, Sigma; 10 pg / ml) plasma (1 :4 diluted or undiluted, VisuCon-F control plasma, Affinity Biologicals Inc) was determined by the time-dependent hydrolysis of fluorogenic substrate. For IC50 and efficacy determination, the compound of Formula A or C1-INH (Sigma Cat #E0518) were added either before (Figures 15a and 15b) or after (Figure 16a) the addition of DXS to the plasma.

[0400] DXS-activated cleavage of HK in undiluted plasma was performed in the absence or presence of 300 nM Pka inhibitor and quantified by SDS-PAGE gel electrophoresis, using 7.5% Criterion TGX Precast gels (Biorad). Transfer was made onto Immunobilon-FL PVDF membrane. Image analysis was done using the LICOR imaging system. Mouse monoclonal anti-HK antibody (MAB15692, R&D systems) was used for traditional immunoblotting. Data presented as % of HK remaining after 20 min incubation with DXS compared to HK levels in unactivated plasma (Table 14).

[0401] Plasma free fraction was determined using “Rapid Equilibrium Dialysis” system (Thermo Scientific), test compounds were prepared at 5 pM in neat human plasma and dialysed against phosphate buffer for 5 hrs at 37°C. Quantification of the compound partitioned in two chambers of the dialysis device was performed via LCMS / MS. Fraction of compound unbound to plasma proteins presented as % of total.

[0402] The ability of the compound to inhibit the enzyme activity of pre-activated plasma was assessed by addition of the compound after DXS stimulation. Aliquots of plasma (20 pL) were mixed with a 2.5 pL solution containing 1 ,300 mM fluorogenic substrate (H-Dpro-Phe-Arg- AFC) and a 2.5 pL solution of dextran sulphate (DXS; 100 pg / mL) which acted as an activator of the plasma kallikrein-kinin pathway. Enzyme activity was immediately measured by monitoring the accumulation of fluorescence liberated from the substrate by substrate cleavage over 16 minutes. At 3.5 minutes post DXS addition 5 pl of inhibitors or water control are were added into each well. The compound was tested at concentrations of 300, 1000 and 3000 nM. C1-INH at a concentration of 3000 nM and vehicle controls were also included. Data are presented in Figure 16b.

[0403] Results:

[0404] As shown in Figures 15a and 15b, in assays using the fluorogenic substrate, the compound of Formula A appears to be a highly potent inhibitor of Pka with 17-fold and 20-fold potency vs. exogenously added C1-INH in diluted plasma (Figure 15a) and undiluted plasma (Figure 15b), respectively.

[0405] Table 14 showing the biochemical profile of the therapies tested in this example.

[0406] Table 14

[0407] Figure 16a shows a comparison of the effects of the two inhibitors: compound of Formula A and C1-INH, on plasma kallikrein activity in plasma (diluted 1 :4) activated with DXS. Both inhibitors were added at concentrations ten times their IC50 to plasma approximately 100 seconds after the addition of the DXS.

[0408] Figure 16b shows that addition of the compound of Formula A after the activation of plasma causes rapid and dose dependent inhibition of enzyme activity compared to the slower action of C1-INH. Table 15 shows the biochemical potency and selectivity of the compound of Formula A against human isolated enzymes using literature methods as for the above-described in vitro plasma kallikrein assay.

[0409] Table 15

[0410] Example 13 - Preparation of a dosage form comprising the compound of Formula A

[0411] Blending and Roller Compaction

[0412] Equipment: Freund Vector TFC Lab Micro Roller Compactor and Granulator (the roller compactor and granulator are separate entities). The equipment parameters are below:

[0413] Table 16

[0414] Method

[0415] Two tablet formulations (Tablets A and B) were prepared according to the following method at 30g blend scale to produce tablets having components in the amounts shown below. Table 17

[0416] For each of the tablets, blends were prepared by passing the intragranular components through a 355 pm sieve at a suitable scale for the scope of the roller compactor in a glass vessel using a Turbula Blender at 34 rpm. The blend was then run through the roller compactor using the parameters described above. The ribbons produced were collected into a suitably sized container. The collected ribbons were then subjected to the granulator fixed with a 1 mm screen and the resultant granules were collected for further downstream processing.

[0417] Tabletting

[0418] Equipment: RIVA Mini single station Tablet Press. The equipment parameters are shown below:

[0419] Table 18

[0420] The granules were subsequently blended with their extragranular excipients, respectively. The extragranular excipients were prepared by screening through a 355 pm sieve in a glass vessel using a T urbula Blender at 34 rpm. The target tablet weight was then dispensed and manually compressed into tablets. Tablet A was compressed at 7.2 to 8.8 kN compression force. Tablet B was compressed at 6.9 to 7.7 kN compression force.

[0421] The tablets were found to be robust. Tablets A and B were subsequently submitted for longterm stability testing.

[0422] The production of tablets per the method described above has been scaled to 180g with a roller compaction time of approximately 60 minutes.

[0423] Aim: To evaluate the pharmacodynamic (PD) effects of the compound of Formula A when orally administered using ex vivo whole plasma assays for plasma kallikrein catalytic activity and HK cleavage, in samples from a Phase 1 Single Ascending Dose Study in healthy adult males. Also, an aim was to investigate safety, tolerability and pharmacokinetic (PK) effects of the compound of Formula A when orally administered.

[0424] Methods:

[0425] This study was a randomized, double-blind, placebo-controlled single ascending dose (SAD) and crossover studies for food effect and capsule / tablet formulations.

[0426] 64 healthy male participants (n=6 active, 2 placebo per cohort, 8 SAD cohorts) were administered single ascending doses of the compound of Formula A: 5, 10, 20, 40, 80, 160, 300 or 600 mg in a capsule.

[0427] 8 participants were administered 100 mg the compound of Formula A in a crossover study of the capsule and a tablet formulation. 12 participants were administered 600 mg the compound of Formula A in a food effect crossover study.

[0428] Samples for pharmacokinetic (PK) and PD assessment were taken at repeated intervals over 48 hours.

[0429] Plasma samples used for PK assessment were analysed using a validated liquid chromatography tandem mass spectrometry (LC MS / MS) method.

[0430] PD measurements were determined in dextran sulfate (DXS) stimulated undiluted plasma using a fluorogenic enzyme assay and capillary based HK cleavage immunoassay.

[0431] Catalytic activity of Pka in DXS-stimulated (Sigma; 10 pg / mL) plasma samples from the compound of Formula A phase 1 study was determined by the time-dependent hydrolysis of fluorogenic substrate in all samples from all parts of the study.

[0432] The time until appearance of detectable amidolytic enzyme activity in DXS-stimulated plasma (lag time) was calculated from the catalytic activity assay. The detection sensitivity of the rate of catalytic activity in plasma based on using a Spark (Tecan) fluorimeter is a fluorescence increase to reach 1AF unit / sec.

[0433] DXS-stimulated cleavage of HK, in undiluted plasma was quantified by capillary-based immunoassay on the Wes System (ProteinSimple) using monoclonal anti-HK antibody and chemiluminescence-based detection. Plasma kallikrein mediated HK cleavage in undiluted citrated human plasma was induced by contact system activation with DXS (6.25 pg / ml) at 4°C in selected samples from the SAD phase.

[0434] DXS-stimulated cleavage of plasma prekallikrein and Factor XII (FXII) were quantified by capillary-based immunoassay on the Wes System (ProteinSimple) analogously.

[0435] Results:

[0436] Figure 17a shows the plasma concentrations of the compound of Formula A from 0 to 24 hours post-dose. As can be seen, when orally administered, the compound of Formula A achieved rapid and dose-dependent plasma exposure over the range of doses tested from 5 mg to 600 mg. Figure 17a shows the concentration curves and Figure 17b shows the Cmax for each SAD cohort. The compound of Formula A was administered as a capsule formulation and the subject was in the fasted state.

[0437] Figure 18a shows enzyme assays in activated undiluted plasma performed on samples from cohorts 6, 7, and 8. Doses 160 mg and above demonstrated >90% average inhibition of plasma kallikrein catalytic activity between 45 min and 2 hr for cohort 6, between 20 min and 4 hr for cohort 7. A 600 mg dose (cohort 8) provided >90% inhibition of plasma kallikrein catalytic activity between 30 min and 6 hr post-dose and >50% inhibition for 10hr (Figure 18b).

[0438] The kinetic fluorescent measurements from the undiluted plasma enzyme assay can be plotted as assay progression curves (Figures 19a and 19b). These curves highlight that the compound of Formula A not only has an inhibitory effect on enzyme activity but also increases the time until appearance of catalytic activity during contact system activation (lag time). At early time points post-dose administration, plasma samples did not display detectable catalytic activity even after prolonged activation with the potent activator DXS. In this test, the subject was administered with 600 mg dose in a tablet formulation.

[0439] Figure 20 shows the mean percent HK protection in DXS-activated undiluted plasma (SAD cohort 6 (160 mg), 7 (300 mg) and 8 (600 mg)). As shown, all three doses of the compound of Formula A were able to inhibit plasma kallikrein catalytic activity above 90% for a period of time. The duration of these PD effects was dose proportional. The compound of Formula A is shown to protect HK from DXS-activated cleavage in undiluted plasma for at least 10 hr following a single 600 mg dose.

[0440] In Figure 20, the representative WES system gel image was generated in duplicate undiluted plasma samples + / - DXS activation from a single subject in cohort 8 who received 600 mg of the compound of Formula A compared with pre-dose (P-D).

[0441] In Figure 20, HK cleavage was evaluated following DXS activation of undiluted plasma samples at selected time points from cohorts 6 to 8. Data are expressed as Mean + / - SEM, n=6.

[0442] To assess whether the compound of Formula A also reduced the generation of plasma kallikrein and Factor XI la, immunoassays were used to quantify levels of contact system proteins in DXS-activated plasma at pre-dose and up to 12 hrs post-dose of 600 mg orally administered in capsules. The results from these assays are shown in Figures 21 to 24 and shows that the compound of Formula A not only reduces HK cleavage but also reduces PPK cleavage and reduces the generation of FXIIa. These results suggest that the compound of Formula A inhibits the contact activation system via interruption of the positive feedback loop mediated by Pka stimulated activation of FXII.

[0443] Figure 25 shows that no significant food-effect was observed on the pharmacodynamic (PD) profile of a 600 mg tablet provided in fed and fasted state. As can be seen, the PD effects are rapidly observed in fed and fasted state with plasma kallikrein inhibition of >90% achieved by 30 minutes in both states.

[0444] No serious adverse events were reported in the phase I trial. There were also no tolerability signals. No subjects withdrew from the trial.

[0445] These data demonstrate that the compound of Formula A has an inhibitory effect on the bradykinin and contact activation systems. As discussed above, these pharmacodynamic effects are implicated in disorders such as HAE. These data also show that the compound of Formula A has a pharmacokinetic profile suitable for oral administration.

[0446] Example 15 - Immunoassays investigating the compound of formula A in the protection of high molecularweight kininogen (HK) from Pka -mediated cleavage in HAE and control plasma

[0447] Method:

[0448] High molecular weight kininogen (HK) cleavage in undiluted citrated human plasma was induced by contact system activation with dextran sulfate (DXS, Sigma #31395-10G; 6.25 pg / ml) on wet ice. Pooled normal (CONTROL) human plasma (VisuCon-F Frozen Normal Control plasma) was purchased from Affinity Biologicals Inc. A working stock of 10mM of the compound of formula A (“the compound”) in DMSO was prepared and diluted in 1X PBS to the respective final concentrations described. HAE plasma was obtained from HAE subjects (n=6) and C1-inhibitor deficiency was confirmed by western blotting. Protection of HK from Pka mediated cleavage in DXS-stimulated whole undiluted plasma was then determined by two methods; traditional western blotting and a semi-automated capillary-based immunoassay.

[0449] Western blotting: SDS-PAGE gel electrophoresis was done using 7.5% Criterion TGX Precast gels (Bio-rad). Transfer was made onto Immobilon-FL PVDF membrane. Image analysis was performed using the LICOR imaging system. Mouse monoclonal anti-human HK antibody (MAB15692, R&D systems) was used for traditional immunoblotting. Capillary-based immunoassay on the WES System (ProteinSimple):

[0450] Preparation of samples: Combine one part 5* fluorescent master mix with four parts of the 1 :200 plasma sample. Vortex to mix. Heat the samples + fluorescent master mix and the biotinylated ladder at 95°C for 5 minutes, vortex, and load onto the WES plate. Monoclonal anti-human HK antibody was used for this chemiluminescence-based detection method using the Wes System (ProteinSimple).

[0451] Analysis: Collect the peak area measurement obtained in the Compass software (cbz file) for the full-length HK molecular weight of the respective time-point sample with DXS-induced activation. The peak area is defined as the area calculated for the spectral peak profile for HK. To measure the plasma kallikrein inhibition by the compound, the percent full-length HK detected was calculated.

[0452] Results:

[0453] Figures 26a and 26b show the time course of dextran sulfate-activated cleavage of HK in HAE whole undiluted plasma determined using western blotting, and a representative blot.

[0454] Figures 27a and 27b shows a representative WES system gel image and that the compound of Formula A provides dose dependent protection against HK cleavage in both HAE and healthy control plasma stimulated with dextran sulfate determined by capillary-based immunoassay using the WES system.

[0455] Example 16 - Phase 2 study of the compound of Formula A

[0456] Aim: To evaluate the efficacy and safety of the compound of Formula A in the on-demand treatment of angioedema attacks in adult subjects with hereditary angioedema type I or II.

[0457] Methods:

[0458] The study was a randomized, double-blind, placebo-controlled, phase 2, cross-over clinical trial evaluating the efficacy and safety of the compound of formula A (“the compound”), an oral plasma kallikrein inhibitor, in the on-demand treatment of angioedema attacks in adult subjects with hereditary angioedema type I or II (EudraCT number: 2018-004489-32).

[0459] Objectives: Primary Objective:

[0460] To investigate the efficacy of the compound compared to placebo in halting the progression of a peripheral or abdominal attack of hereditary angioedema (HAE).

[0461] Secondary Objectives:

[0462] To investigate the safety and tolerability of the compound.

[0463] To investigate the pharmacokinetic (PK) profile of the compound when taken during the intercritical period between HAE attacks.

[0464] To investigate the pharmacodynamic (PD) profile of the compound in reducing the concentration of residual cleaved high molecular weight kininogen (HK) during the intercritical period between HAE attacks.

[0465] To investigate the PD profile of the compound in reducing activated plasma enzyme activity during the intercritical period between HAE attacks.

[0466] Setup:

[0467] This was a phase 2, two-part, two-sequence, two-period (2x2) cross-over clinical trial. Subjects with HAE type I or II were recruited through HAE treatment centres in Europe and US.

[0468] In Part 1 , subjects received a single oral dose of 600 mg of the compound to investigate the safety, PK and PD of the compound during the intercritical period between HAE attacks.

[0469] Eligible adult subjects >18 years old underwent a screening assessment for study inclusion and to receive study drug, followed by a 4h, in-clinic, safety and PK / PD assessment.

[0470] In Part 2, the subjects were randomized 1 :1 to 2 treatment sequences. This part of the study was conducted away from the clinic or hospital. In Sequence 1 (study arm 1) subjects received a single dose of 600 mg of the compound to treat the first eligible HAE attack. Following resolution of this attack, subjects received a single dose of placebo to treat the second eligible HAE attack.

[0471] In Sequence 2 (study arm 2) subjects received a single dose of placebo to treat the first eligible HAE attack. Following resolution of this attack, subjects received a single dose of 600 mg of the compound to treat the second eligible HAE attack.

[0472] A minimum of 48-hour washout period was required between each dose of study drug. Laryngeal or facial attacks were not eligible for treatment. HAE attacks must have been treated within the first hour of onset and before reaching severe on the global attack severity scale. Subjects must also have been able to identify the start of a HAE attack. Upon onset of the eligible HAE attack, subjects notified the dedicated study physician or qualified designee with a description of the HAE attack. The dedicated study physician or qualified designee confirmed eligibility of the HAE attack and agreed to study drug being administered. HAE attacks required documentation, on the Subject Diary, of attack location, attack symptoms, time of onset, attack severity, and time of last substantial meal prior to dosing. Subjects took study drug, as instructed, and completed timed assessments of their HAE attack symptoms for a 48h period as documented below in Table 19. The dedicated study physician or qualified designee contacted the subject within 24h of the eligible HAE attack to confirm the subject’s safety and wellbeing. Subjects were instructed to contact the dedicated study physician or qualified designee in case of any safety concerns. In the case of hypersensitivity, subjects contacted the dedicated study physician or qualified designee or contact the nearest emergency service. The dedicated study physician or qualified designee was available 24h / day and 7 days / week to receive subject calls.

[0473] Table 19

[0474] *ln the event that conventional attack treatment was used, the subject performed assessments every 30 min for 4 h following first administration of conventional attack treatment. After this, the subject reverted back to original frequency of assessments based on time of study drug administration.

[0475] Subjects returned to the clinic following the first HAE attack, prior to the second HAE attack, to undergo safety checks including adverse event (AE) reporting, vital sign recording, and Subject Diary review. Once two HAE attacks were treated in Part 2, the subject returned to the clinic to undergo final safety checks including AE reporting, vital sign recording and blood sampling for laboratory safety measurements.

[0476] Conventional attack treatment was permitted after 4h, or earlier as warranted, following study drug intake, provided HAE attack symptoms were judged severe enough by the subject to require treatment as per the subject’s usual treatment regimen, or are deemed ineligible for study drug treatment, or were associated with laryngeal or facial symptoms. Prior to use of conventional attack treatment, subjects notified the dedicated study physician or qualified designee who confirmed conventional treatment was appropriate per protocol and subject report of symptom severity. Subjects were permitted to treat their HAE attacks with their conventional attack treatment (pdCIINH or rhCHNH intravenous [iv] or icatibant).

[0477] Investigational Medicinal Product:

[0478] The compound of formula A - 100 mg film-coated tablet. These contained the following excipients: microcrystalline cellulose, croscarmellose sodium, povidone, magnesium stearate; the aesthetic coating contains hypromellose, lactose monohydrate, titanium dioxide and tri acetin.

[0479] Placebo to the compound 100 mg film-coated tablet. These contained microcrystalline cellulose, colloidal silicon dioxide, sodium starch glycolate, and sodium stearyl fumarate and are film-coated; the aesthetic coating contains hypromellose, lactose monohydrate, titanium dioxide and triacetin.

[0480] No study drug dose modifications were allowed in this study.

[0481] Number of Subjects: 68 subjects were enrolled into the study to ensure 50 subjects complete the study. 53 subjects completed Part 2.

[0482] Population: The study population included male and female subjects 18 years of age or older with HAE type I or II.

[0483] Inclusion Criteria:

[0484] 1. Male or female adult subjects 18 years of age and older.

[0485] 2. Confirmed diagnosis of HAE type I or II at anytime in the medical history: a. Documented clinical history consistent with HAE (subcutaneous or mucosal, nonpruritic swelling episodes without accompanying urticaria) AND b. C1-esterase inhibitor (C1-INH) antigen or functional level <40% of the normal level. Subjects with antigen or functional C1-INH level 40-50% of the normal level were enrolled if they also have a C4 level below the normal range and a family history consistent with HAE type I or II.

[0486] 3. At least 3 documented HAE attacks in the past 93 days, as supported by medical history.

[0487] 4. Access to and ability to use conventional attack treatment for attacks of HAE.

[0488] 5. Adequate organ functions as defined below: a. Hemoglobin within normal range; b. International normalized ratio (INR)< 1.2; c. Activated partial thromboplastin time (aPTT) < upper limit of normal (ULN); d. Creatinine < 1x ULN; e. Creatinine clearance (CrCI) > 60 mL / min; f. Alanine aminotransferase (ALT) < 2x ULN; g. Aspartate aminotransferase (AST) < 2x ULN; h. Total bilirubin < 1.5x ULN; i. Leucocytes < 1.5x ULN; j. Thrombocytes < 1.5x ULN.

[0489] 6. Female of childbearing potential must have agreed to use highly effective birth control from the Screening visit until the end of the trial follow-up procedures.

[0490] Highly effective methods of birth control include: a. Progestogen-only hormonal contraception associated with inhibition of ovulation: oral I injectable I implantable.

[0491] (Hormonal contraception that contains estrogen was excluded per exclusion criterion 3). b. Intrauterine device (IUD). c. Intrauterine hormone-releasing system (IUS). d. Bilateral tubal occlusion. e. Vasectomised partner (provided that the partner was the sole sexual partner of the female subject of childbearing potential and that the vasectomised partner received medical assessment of the surgical success). f. Sexual abstinence (this method is not acceptable in Switzerland).

[0492] Note: Sexual abstinence was only be considered a highly effective method if it is defined as refraining from heterosexual intercourse. The reliability of sexual abstinence needed to be evaluated in relation to the duration of the clinical trial and the preferred and usual lifestyle of the subject.

[0493] 7. Females of non-childbearing potential, defined as surgically sterile (status post hysterectomy, bilateral oophorectomy, or bilateral tubal ligation) or post-menopausal for at least 12 months, did not require contraception during the study.

[0494] 8. Males with female partners of childbearing potential must have agreed to be abstinent or else use a highly effective method of birth control as defined in inclusion criterion 6 from the Screening visit until the end of the trial follow-up procedures.

[0495] 9. Provide signed informed consent and were willing and capable of complying with study requirements and procedures.

[0496] Exclusion criteria:

[0497] 1. Any concomitant diagnosis of another form of chronic angioedema, such as acquired C1 inhibitor deficiency, HAE with normal C1-INH (also known as HAE type III), idiopathic angioedema, or angioedema associated with urticaria.

[0498] 2. Current use of C1 INH, androgens, lanadelumab or tranexamic acid for HAE prophylaxis.

[0499] 3. Use of angiotensin-converting enzyme (ACE) inhibitors or any estrogen-containing medications with systemic absorption (such as oral contraceptives or hormonal replacement therapy) within 93 days prior to initial study treatment.

[0500] 4. Use of androgens (e.g. stanozolol, danazol, oxandrolone, methyltestosterones, testosterone) or antifibrinolytics within 30 days prior to initial study treatment.

[0501] 5. Use of lanadelumab within 10 weeks prior to initial study treatment.

[0502] 6. Use of strong CYP3A4 / CYP2C9 inhibitors and inducers during participation in the trial.

[0503] Note: These medications include but are not limited to the following: cobicistat, conivaptan, itraconazole, ketoconazole, posaconazole, voriconazole, ritonavir, boceprevir, telaprevir, troleandomycin, clarithromycin, carbamazepine, enzalutamide, mitotane, phenytoin, phenobarbital, fluconazole, isoniazid, metronidazole, paroxetine, sulfamethoxazole, rifampicin, St. John’s Wort, diltiazem, idelalisib, nefazodone and nelfinavir.

[0504] 7. Clinically significant abnormal electrocardiogram (ECG) at Visit 1 and pre-dose at Visit 2. This includes, but is not limited to, a QTcF > 470 msec (for women) or > 450 msec (for men), a PR > 220 msec or ventricular and / or atrial premature contractions that are more frequent than occasional and / or occur as couplets or higher in grouping. 8. Any clinically significant history of angina, myocardial infarction, syncope, clinically significant cardiac arrhythmias, left ventricular hypertrophy, cardiomyopathy, or any other cardiovascular abnormality.

[0505] 9. Any other systemic dysfunction (e.g., gastrointestinal, renal, respiratory, cardiovascular) or significant disease or disorder which, in the opinion of the Investigator, would have jeopardized the safety of the subject by taking part in the trial.

[0506] 10. History of substance abuse or dependence that would interefere with the completion of the study, as determined by the Investigator.

[0507] 11. Known lactose allergy or intolerance.

[0508] 12. Known hypersensitivity to the compound or placebo or to any of the excipients.

[0509] 13. Participation in an interventional investigational clinical study within 93 days or within 5 half-lives of the last dosing of investigational drug (whichever is longer) prior to initial study treatment.

[0510] 14. Any pregnant or breast-feeding subject.

[0511] Assessments:

[0512] Part 1: Blood samples for PK and PD measurements were collected at the following timepoints: Pre-dose (Oh), 15 min, 30 min, 45 min, 1 h, 1.5h, 2h, 3h, and 4h post-dose. Vital signs (systolic blood pressure [SBP], diastolic blood pressure [DBP], pulse rate [PR], respiratory rate [RR] and body temperature) were measured at pre-dose (Oh), 1h, and 4h post-dose. Samples for post- treatment safety laboratory assessments were taken with the 4h PK / PD samples.

[0513] Part 2: Following study drug intake, subject assessments of overall HAE attack severity and change in HAE attack severity took place for a 48h period as documented in Table 19 above.

[0514] Efficacy Variables:

[0515] Time to use of conventional attack treatment was assessed. The subject diary captured the efficacy endpoints including time to use of conventional attack treatment and HAE attack severity.

[0516] Overall HAE attack severity was assessed on a 5-point Likert scale (5LS) scored as none, mild, moderate, severe and very severe. Change in HAE attack severity was assessed using a 7-point transition question (7TQ), scored as Much better / Better / A little better / No change / A little worse / Worse / Much worse.

[0517] The type of HAE attack symptoms (abdominal pain, skin pain and skin swelling) was each assessed on a 100 mm visual analogue scale (VAS) anchored at 0 (none) and 100 (very severe).

[0518] Safety Variables:

[0519] Aes, including serious adverse events (SAEs).

[0520] Laboratory test results (clinical chemistry, hematology, coagulation, and urinalysis).

[0521] Vital signs (SBP, DBP, PR, RR, body temperature).

[0522] Physical examination findings.

[0523] ECG results.

[0524] Pregnancy test (female subjects of child-bearing potential).

[0525] Criteria for Evaluation of Efficacy

[0526] Primary Efficacy Endpoints:

[0527] Time to use of conventional attack treatment.

[0528] Secondary Efficacy Endpoints:

[0529] Proportion of HAE attacks that progress by one level or more on the 5LS or that require conventional attack treatment within 12h of study drug.

[0530] Time between treatment and (1) progression of global attack severity on the 5LS by one level or more, or (2) use of conventional attack treatment, whichever comes first within 12h.

[0531] Exploratory Endpoints:

[0532] Cumulative global attack severity on the 5LS following study drug expressed as area under the curve (AUC) for the compound 600 mg vs. placebo.

[0533] Proportion of HAE attacks that require conventional attack treatment.

[0534] Proportion of HAE attacks that are rated “worse” or “much worse” on the 7TQ.

[0535] Proportion of HAE attacks that are rated “better” or “much better” on the 7TQ.

[0536] Time from study drug administration to complete HAE attack resolution (rating of none) on global attack severity scale (5LS).

[0537] Time to HAE attack being rated worse or much worse on the 7TQ.

[0538] Time to HAE attack being rated better or much better on the 7TQ. General Statistical Methods and Types of Analyses

[0539] Analysis Sets:

[0540] Safety set (SAF): Subjects who have taken at least one dose of study drug (including the study drug dose in Part 1).

[0541] Full analysis set (for efficacy) (FAS): All randomized subjects who received both doses of study drug in Part 2.

[0542] Per protocol set (for efficacy) (PPS): Randomized subjects in Part 2 who received both doses of study drug (the compound) in Part 2 and have no major protocol deviations.

[0543] PK I PD analysis set: All subjects for whom PK I PD samples were taken in Part 1 .

[0544] Sample size:

[0545] A sample size of 50 subjects (25 per sequence) was proposed to provide 90% power for testing at the 5% alpha level (2-sided) for the primary endpoint of time to use of conventional attack treatment. This sample size was derived based upon an assumption that 40% of subjects will use conventional attack treatment while on the control arm while 10% will use conventional attack treatment on the experimental arm and that within subject data has minimal correlation. The assumption of minimal correlation should be a conservative assumption with respect to sample size. 68 subjects were enrolled to ensure that 50 subjects complete the study. 53 subjects completed part 2.

[0546] An oversampling by 20% (10 subjects) was proposed to account for subjects that may not complete both treatment periods due to infrequent or ineligible HAE attacks or for subjects who discontinue the trial early, for whatever reason. Thus, study enrolment was considered sufficient to address the primary efficacy hypothesis after 50 subjects have completed both treatment periods. Since further exposure was not required and could be considered unnecessary, ongoing subjects who have not completed both periods were asked to return to the study site and complete Visit 4 (Early Discontinuation visit). Data from all subjects, complete and incomplete, was analyzed in the safety set.

[0547] General Considerations:

[0548] Individual subject data was presented in subject data listings. Appropriate descriptive statistics was calculated for continuous and categorical data and summarized in tabular format.

[0549] Sample Analyses: Aes were coded using the Medical Dictionary for Regulatory Activities (MedDRA) dictionary (v21.0 or higher) and classified by preferred term and system organ class (SOC). Listings of treatment-emergent adverse events (TEAEs), serious TEAEs, and TEAEs causing premature discontinuation will be provided by sequence group, and further classified by TEAE severity and relationship to study drug.

[0550] Efficacy Analyses:

[0551] Primary Endpoint

[0552] The primary endpoint, time to use of conventional attack treatment, was analyzed using a generalization of Gehan’s test proposed by Feingold and Gillespie (1996) (Crossover trials with censored data. Statistics in Medicine 1996; 15(10): 953-967) to reflect the repeat measures on each subject. Subjects were treated as censored if no worsening occurs within 12h of study drug.

[0553] Secondary Endpoints

[0554] The proportion of HAE attacks that worsen by one level or more on the 5LS or that require conventional attack treatment within 12h of study drug was analyzed using Prescott’s test (1981) (The comparison of success rates in cross-over trials in the presence of an order effect. Applied Statistics 1981 ; 30: 9-15) to compare the treatment arms.

[0555] A similar approach to that used for the primary endpoint was followed for the analysis of the time between study drug and HAE attack worsening by one level or more on the 5LS or use of conventional attack treatment, whichever comes first within 12h. In addition to the tests described above, descriptive statistics were presented for the primary, secondary and exploratory endpoints, in each case comparing the compound to placebo, such as:

[0556] Cumulative global attack severity on the 5LS following study drug expressed as AUC for the compound 600 mg vs. placebo.

[0557] Proportion of HAE attacks that required conventional attack treatment.

[0558] Proportion of HAE attacks that were rated “worse” or “much worse” on the 7TQ.

[0559] Proportion of HAE attacks that were rated “better” or “much better” on the 7TQ.

[0560] Time from study drug administration to complete HAE attack resolution (rating of none) on global attack severity scale (5LS).

[0561] Time to HAE attack being rated worse or much worse on the 7TQ.

[0562] Time to HAE attack being rated better or much better on the 7TQ. PK Analysis:

[0563] Non-compartmental PK parameters included maximum concentration in plasma (Cmax), time to reach Cmax in plasma (tmax), and area under the curve from time 0 to last sample (AUC0- t). Compartmental PK modelling described the PK of the compound and generate underlying Cmax, tmax, AUC, apparent clearance (CL / F), apparent volume of distribution (Vd / F) and estimated terminal elimination half-life

[0564] The PK parameters of the compound were determined from the individual concentration versus time data using Phoenix WinNonlin. In case of a deviation from the theoretical time, the actual time of blood sample was used in the calculation of the derived PK parameters. Individual concentrations and derived PK parameters of the compound in plasma were listed and summarized for each treatment. Individual and geometric mean concentration-time data were plotted on linear and semi-logarithmic scales.

[0565] PD Analysis:

[0566] The compound’s effect on plasma kallikrein (Pka) activity was analyzed using two exploratory measures of Pka enzyme activity in plasma:

[0567] An assay to determine inhibition of exogenously activated plasma kallikrein enzyme activity from plasma samples obtained before and after receiving the compound.

[0568] An assay to measure the level of protection of cleavage of high molecular weight kininogen (HK) substrate (contained in whole plasma) from plasma kallikrein enzyme activity.

[0569] The PD were summarized for each treatment. Individual and mean data were provided as a report addendum located in the appendix of the final Clinical Study Report.

[0570] Preliminary PK data from Part 1 of the study:

[0571] Preliminary PK data from 27 HAE patients was collated and analysed, and are shown in Table 20 and Figure 28a.

[0572] Table 20

[0573] Thus, these preliminary results show that the compound of Formula A demonstrates a pharmacokinetic profile that is suitable for on-demand oral administration in HAE patients.

[0574] Figure 28b shows the complete PK profile of HAE patients from part 1 , overlaid with PK data from healthy volunteers.

[0575] Efficacy results from Part 2 of the study

[0576] Primary Endpoint

[0577] Attacks treated with the compound significantly reduced use of rescue (p=0.0010) with 15.1% of compound treated attacks rescued compared to 30.2% on placebo at 12 hours. This efficacy benefit of the compound was maintained at 24 hours (p=0.0005). At 24 hours, use of rescue medication (p=0.0005) was 20.8% for the compound versus 39.6% on placebo.

[0578] Figure 28c shows the cumulative % of patients that used rescue medication within 12 hours of being administered the compound.

[0579] Secondary Endpoints

[0580] The compound significantly reduced time to onset of symptom relief (p=<0.0001) on a Patient Global Impression of Change scale (PGI-C) (also known as the 7TQ) with a median time of 1.6 hours versus 9 hours for attacks treated with placebo. This demonstrates that treatment with the compound achieved symptom relief more quickly than placebo.

[0581] Within 12 hours of study drug, 83.0% of attacks treated with the compound were rated “a little better” or higher (when assessed according to the 7TQ), compared to 50.9% of attacks treated with Placebo (p<0.005). Within 24 hours of study drug, 84.9% of attacks treated with the compound were rated “a little better” or higher (when assessed according to the 7TQ), compared to 64.2% of attacks treated with Placebo (p<0.05).

[0582] Within 12 hours of study drug, 58.7% of attacks treated with the compound were rated “better” or higher (when assessed according to the 7TQ), compared to 35.8% of attacks treated with placebo (p = 0.0319). Within 24 hours of study drug, 67.9% of attacks treated with the compound were rated “better” or higher (when assessed according to the 7TQ), compared to 47.2% of attacks treated with placebo (p = 0.0593).

[0583] T reatment with the compound led to a statistically significant shortening of the median time for attacks to be rated “better” or higher (when assessed according to the 7TQ), with a median time of 5 hours for treatment with the compound (p = 0.0003) compared to a median time of 15 hours for placebo (p = 0.0036).

[0584] Of the attacks that achieved a rating of “better” or higher within 24 hours (when assessed according to the 7TQ), 6.1% used rescue medication, 66.7% achieved attack resolution when assessed according to the PGI-S, and 77.6% achieved attack resolution when assessed by VAS. On the contrary, of the attacks that did not achieve a rating of “better” or higher within 24 hours (when assessed according to the 7TQ), 63.8% used rescue medication, 4.3% achieved attack resolution when assessed according to the PGI-S, and 5.1% achieved attack resolution when assessed by VAS.

[0585] Figure 28d shows the cumulative % of patients with symptom relief (as assessed using the 7TQ method) within 12 hours of being administered the compound. Figure 28e compares time to symptom relief data for the compound with known clinical data for approved injectable product Ruconest® (see Charles M. Maplethorpe, MD, PhD. Clinical Reviewer. Summary Basis of Approval, Recombinant C1 Esterase Inhibitor, STN: 125495 / 0).

[0586] Attacks treated with the compound achieved symptom relief more quickly than placebo treated attacks (p<0.0001) when assessed using a composite Visual Analogue Scale (VAS) score (abdominal pain, skin pain, and skin swelling) with a median time to symptom relief of 6 hours versus >12 hours for placebo. Again, this demonstrated that treatment with the compound achieved symptom relief more quickly than placebo.

[0587] Figure 28f shows the cumulative % of patients with symptom relief (as assessed using the composite VAS score) within 12 hours of being administered the compound.

[0588] Attacks treated with the compound reduced the severity of the HAE attack (as assessed using the composite VAS score) over 12 hours (p=0.0008) and 24 hours (p=0.0005). Within 12 hours of study drug, 79.2% of attacks treated with the compound do not worsen, compared to 54.7% of attacks treated with Placebo. Figure 28g shows the mean composite VAS score within 12 hours of being administered the compound. Figure 28g is time- and baseline- adjusted AUC and excludes assessments postrescue medication. Figure 28h compares the mean composite VAS score for patients treated with the compound with known clinical data for approved injectable product Firazyr® (active ingredient, icatibant) (see Lumry et al., Ann Allergy Asthma Immunol. 2011 ;107:529 -537).

[0589] Within 12 hours of oral administration, the compound significantly increased the number of attacks stabilized or improved when assessed by a Patient Global Impression of Severity scale (PGI-S) (also known as the 5LS) or use of rescue (p<0.0001).

[0590] Figure 28i shows the % of HAE attacks that did not worsen (as assessed using the 5LS method) within 12 hours of being administered the compound. As noted above, within 12 hours of study drug, 79.2% of attacks treated with the compound do not worsen, compared to 54.7% of attacks treated with Placebo. Within 24 hours of study drug, 69.8% of attacks treated with the compound do not worsen, compared to 50.9% of attacks treated with Placebo.

[0591] Within 12 hours of study drug, 37.7% of attacks treated with the compound improved to “none” when assessed using the PGI-S (also called the 5LS method), compared to 18.9% of attacks treated with Placebo. Within 24 hours of study drug, 52.8% of attacks treated with the compound improved to “none”, compared to 26.4% of attacks treated with Placebo.

[0592] Results from other measured endpoints were as follows:

[0593] Table 21

[0594] Of the attacks treated with the compound of Formula A, 31.0% and 69.0% were categorized as abdominal and peripheral attacks, respectively, and 77.8% of abdominal attacks and 77.5% of peripheral attacks achieved symptom relief (PGI-C score of at least “A little better” within 12 hours). Median time to symptom relief was 1.5 hours for abdominal attacks and 2.5 hours for peripheral attacks). Baseline attack severity was 1.7 for abdominal and 1.5 for peripheral, where “baseline attack severity” is assessed by assigning numeric values from 0 to 4 to align with categorical PGI Severity scale (PGI-S) scores (from “None” to “Very Severe”). A total of 61 .1 % of abdominal attacks and 50.0% of peripheral attacks achieved attack resolution within 24 hours.

[0595] Safety

[0596] There were no serious adverse events reported in the trial and no patients withdrew due to adverse events. In the open-label phase (Part 1), 8 on-treatment drug-related treatment emergent adverse events (TEAE) were experienced by 5 patients. In the crossover phase of the trial (Part 2), 3 on-treatment drug-related TEAEs were experienced by 3 patients (5.2%) following administration of the compound, and 2 on-treatment drug-related TEAEs were experienced by 2 patients (3.6%) following administration of placebo. These data are shown below.

[0597] Table 22

[0598] Example 17 - Phase 3 study of the compound of Formula A

[0599] A Randomized, Double-Blind, Placebo-Controlled, Phase 3, Three-way Crossover Trial to Evaluate the Efficacy and Safety of Two Dose Levels of the compound of Formula A (“the compound” or “study drug”), an Oral Plasma Kallikrein Inhibitor, for On-Demand Treatment of Angioedema Attacks in Adolescent and Adult Patients with Hereditary Angioedema Type I or II.

[0600] Objectives:

[0601] Primary Objective:

[0602] To demonstrate the clinical efficacy of the compound compared with placebo for the on-demand treatment of HAE attacks.

[0603] Secondary Objective:

[0604] To investigate the safety and tolerability of the compound.

[0605] Setup:

[0606] This will be a double-blind, randomized, placebo-controlled, multicentre clinical trial in patients 12 years or older with HAE type I or II. Patients will be randomized to 6 treatment sequences in a 3-way crossover design. Eligible attacks will initially be treated with a single dose of placebo, 300 mg, or 600 mg of the compound per attack with a minimum 48-hour washout period between each eligible attack and last dose of compound or conventional on-demand treatment. If needed (as determined by the patient), a second dose of the study drug may be administered for each attack.

[0607] The estimated duration of this trial for each randomized patient will be approximately 25 weeks from screening through the final visit and includes the treatment of 3 eligible attacks during the treatment period. This trial will be conducted at HAE treatment centres on an outpatient basis and will comprise in-clinic and televisits. A televisit can be conducted via a telephone call or a via an interactive audio / video system. If an in-clinic visit cannot be conducted (e.g. in the event of a pandemic or other reason that prevents the patient from attending the in-clinic visit) home health visits will be used to perform these visits if permitted by the relevant regulatory authority, site’s Ethics Committee (EC) / Institutional Review Board (I B), local regulations, and the patient via informed consent. The home visit will be performed by an appropriately delegated home healthcare service provider. Information captured during a home health visit will mirror that captured in an in-clinic visit.

[0608] Screening Visit

[0609] Eligible patients >12 years old will undergo a screening assessment for trial inclusion. All patients will provide informed consent or assent prior to any trial-related procedures being performed. Informed consent and assent may be collected through e-consent if allowed through country and site regulations.

[0610] During the visit, a physical exam, 12-lead electrocardiogram (ECG), laboratory tests (including diagnostic testing for HAE), and other assessments will be performed.

[0611] Site personnel will train patients on the information they will be expected to provide in the electronic diary (eDiary) and the use of the study drug.

[0612] Randomization Visit

[0613] Within 4 weeks of the Screening Visit, patients will participate in a Randomization Visit. Patients will be assigned to receive 3 treatments in randomized, double-dummy blinded, crossover fashion based on their assignment to 1 of 6 treatment sequences. Randomization will be stratified by whether the patient enters the trial taking only conventional on-demand treatment vs. on a stable dose and regimen of long-term prophylactic treatment. Randomization will occur in a 1 : 1 : 1 : 1 : 1 : 1 ratio using a permuted-block randomization method to ensure a balanced assignment to each treatment sequence. Each patient will receive the following treatments:

[0614] 300 mg of the compound (1 x 300 mg tablet plus 1 matching placebo tablet)

[0615] 600 mg of the compound (2 x 300 mg tablets) 2 matching placebo tablets

[0616] Patients will treat each eligible attack with up to 2 doses of the study drug, administered at least 3 hours apart. The second dose, if taken, will be the same assigned treatment as the first dose.

[0617] In this trial, the study drug will be shipped directly to the patients via a courier service or will be dispensed at the trial clinic as required by local regulations or per the site’s local practice, as described in the Pharmacy Manual.

[0618] Treatment of Eligible HAE Attacks

[0619] Patients will treat 3 separate, eligible HAE attacks with their assigned study drug treatment for that attack. For an HAE attack to be considered eligible for treatment with the study drug, the attack must meet the following criteria:

[0620] • The attack is not a severe laryngeal attack.

[0621] • Patient must be able to identify the start time of the attack.

[0622] • At least 48 hours have elapsed since patient has used conventional on-demand treatment or the study drug to treat an HAE attack.

[0623] • Patient must be able to complete at least the first 4 hours of eDiary assessments following the first administration of the study drug.

[0624] • Post-attack televisit has been completed for the previous eligible attack (applicable to eligible attacks 2 and 3 only).

[0625] Eligible attacks should initially be treated with a single administration of the study drug. Patients will be encouraged to treat as soon as possible after the start of the attack.

[0626] If needed (as determined by the patient), a second dose of the study drug may be administered for each attack, as follows:

[0627] Non-laryncieal attacks

[0628] For each eligible HAE attack, a second dose of the study drug may be taken:

[0629] • after 3 hours if HAE attack symptoms are considered severe enough by the patient to require a second dose of the study drug.

[0630] After the second dose of the study drug, conventional on-demand treatment may be taken: after 1 hour if HAE attack symptoms are considered severe enough by the patient to require treatment with conventional treatment.

[0631] If symptoms progress to airway involvement, patients may treat with conventional on-demand treatment at any time.

[0632] Laryngeal attacks

[0633] After the first dose of the study drug, conventional on-demand treatment may be taken at any time:

[0634] • if HAE attack symptoms worsen or if HAE attack symptoms are considered severe enough by the patient to require immediate treatment.

[0635] Attacks that do not meet eligibility may be treated with conventional on-demand treatment per the patient’s usual treatment regimen.

[0636] Conventional on-demand treatments may include plasma derived C1-inhibitor (pdC1-INH) intravenous (iv), recombinant human C1-esterase inhibitor (rC1-INH) iv, icatibant sc, or ecallantide sc.

[0637] Patients with safety or tolerability concerns will contact the Investigator or designee as soon as possible or the nearest emergency service as appropriate.

[0638] Study Call Center

[0639] After the first dose of the study drug and prior to the second dose or conventional on-demand treatment, patients will be required to call a designated Study Call Center to be reminded of the repeat dosing criteria. Patients are to contact the Call Center:

[0640] 1. After the first dose of the study drug

[0641] 2. Prior to a second dose of the study drug

[0642] 3. Prior to a dose of conventional on-demand treatment.

[0643] The Call Center staff will remind patients of the rules for re-dosing and the eDiary assessment requirements. The Call Center will not collect any data during the call.

[0644] Patient eDiary For each HAE attack treated with the study drug, patients will record information in an eDiary, including attack location, attack symptoms, date / time of onset, attack severity, time of second study drug dose, if applicable, and use of conventional on-demand treatment, if applicable. Patients will complete timed assessments of their HAE attack through 48 hours as documented in Table 23: Frequency of Patient Assessment. Patients should complete all timed diary assessments except during sleep; however, patients must complete at least the first 4 hours of diary assessments following the first administration of the study drug.

[0645] Table 23: Frequency of Patient Assessment

[0646] * If a second dose of IMP or additional doses of conventional on-demand treatment are needed, patients will complete diary assessments prior to taking each additional dose. After re-dosing, the planned post-dose diary assessments will then continue through 48 hours after first dose of the study drug.

[0647] Post-Attack Televisit

[0648] A televisit (between the site staff and the patient) will be completed following each administration of the study drug to ensure the safety and wellbeing of the patient, to confirm the study drug accountability, to review the patient diary (and retrain, if necessary), and to undergo an adverse event (AE) and concomitant medication review. The televisit will occur by the next working day after the completion of the patient diary (visit window: +1 week).

[0649] Final Visit / Early Termination

[0650] Once 3 HAE attacks have been treated, or upon early termination, the patient will return to the clinic as soon as possible within 1 week for an in-clinic visit to undergo final safety checks including AE reporting, vital sign recording, and blood sampling for laboratory safety measurements. Whenever possible, this visit should be completed prior to starting any new medication or treatment. Investigational Medicinal Product:

[0651] The compound of Formula A - 300 mg film-coated tablet. These contained the following excipients: microcrystalline cellulose, croscarmellose sodium, povidone, magnesium stearate; the aesthetic coating contains polyvinyl alcohol, titanium dioxide and polyethylene glycol 3350.

[0652] Placebo for the compound tablet. These contained microcrystalline cellulose, croscarmellose sodium, povidone, magnesium stearate and are film-coated; the aesthetic coating contains polyvinyl alcohol, titanium dioxide and polyethylene glycol 3350.

[0653] No study drug dose modifications will be allowed in this study.

[0654] Tablets must be swallowed whole and are not to be crushed or modified in any way. Eligible attacks should initially be treated with a single administration of the study drug as soon as possible after the start of the attack. If needed (as determined by the patient), a second dose of the study drug may be administered for each attack.

[0655] Number of Patients:

[0656] Approximately 114 patients will be enrolled into the trial to ensure approximately 84 patients (including a minimum of 12 adolescents) complete the trial.

[0657] The trial population will comprise 2 subsets: (1) patients who enter the trial taking only conventional on-demand treatment and (2) patients who enter the trial on a stable dose and regimen of long-term prophylactic treatment

[0658] Population:

[0659] The trial population will include male and female patients 12 years of age and older with a confirmed diagnosis of HAE type I or II.

[0660] The trial population will include a subset of patients who enter the trial taking only conventional on demand treatment and a subset of patients who enter the trial on a stable dose and regimen of long-term prophylactic treatment.

[0661] Inclusion Criteria: 1. Male or female patients 12 years of age and older.

[0662] 2. Confirmed diagnosis of HAE type I or II at any time in the medical history: a. Documented clinical history consistent with HAE (sc or mucosal, nonpruritic swelling episodes without accompanying urticaria) and either: i. Diagnostic testing results obtained during the Screening Period that confirm HAE type I or II: C1-INH functional level <40% of the normal level. Patients with functional C1-INH level 40-50% of the normal level may be enrolled if they also have a C4 level below the normal range. Patients may be retested during the Screening Period if results are incongruent with clinical history or believed by the Investigator to be confounded by recent prophylactic or therapeutic C1-INH use, or ii. Documented genetic results that confirm known mutations for HAE type I or II.

[0663] 3. Patient has access to and ability to use conventional on-demand treatment for HAE attacks.

[0664] 4. If a patient is receiving long-term prophylactic treatment with one of these medicines indicated for HAE: intravenous (iv) or sc plasma-derived C1-INH, and / or lanadelumab, they must be on a stable dose and regimen for at least 3 months prior to the Screening Visit and be willing to remain on a stable dose and regimen for the duration of the trial.

[0665] 5. Patient’s last dose of attenuated androgens was at least 28 days prior to randomization.

[0666] 6. Patient has had at least 2 documented HAE attacks within 3 months prior to randomization.

[0667] 7. Patients must meet one of the following contraception requirements as follows: a. Female patients who are fertile and heterosexual ly active must agree to use contraception from the Screening Visit until the Final or Early Termination (ET) Visit. Acceptable methods of contraception include one or more of the following: i. Progestogen-only hormonal contraception associated with inhibition of ovulation: oral / injectable / implantable (hormonal contraception that contains estrogen including ethinylestradiol is excluded per Exclusion 4). ii. Intrauterine device. iii. Intrauterine hormone-releasing system. iv. Bilateral tubal occlusion. v. Vasectomized partner (provided that the partner is the sole heterosexual partner of the female patient of childbearing potential and that the vasectomized partner has received medical assessment of surgical success). vi. Male or female condom. vii. Cap, diaphragm, or sponge with spermicide. b. Patients who are not fertile or not heterosexually active, as defined below, do not require contraception. If the patient’s status changes during the course of the trial, they will be required to meet the requirements specified in Inclusion Criterion 7a. i. Female patients who refrain from heterosexual intercourse during the trial if the reliability of the heterosexual abstinence has been evaluated in relation to the duration of the clinical trial and is the preferred and usual lifestyle of the patient. ii. Female patients who are surgically sterile (e.g. status post hysterectomy, bilateral oophorectomy, or bilateral tubal ligation) or post-menopausal for at least 12 months. iii. Female patients who are premenarche and remain premenarcheal until the end of the trial. iv. Male patients (including female partners).

[0668] 8. Patients must be able to swallow trial tablets whole.

[0669] 9. Patients, as assessed by the Investigator, must be able to appropriately receive and store the study drug, and be able to read, understand, and complete the electronic diary (eDiary).

[0670] 10. Investigator believes that the patient is willing and able to adhere to all protocol requirements.

[0671] 11 . Patient provides signed informed consent or assent (when applicable). A parent or legally authorized representative must also provide signed informed consent when required.

[0672] Exclusion criteria:

[0673] 1. Any concomitant diagnosis of another form of chronic angioedema, such as acquired Ciinhibitor deficiency, HAE with normal C1-INH (previously known as HAE type III), idiopathic angioedema, or angioedema associated with urticaria.

[0674] 2. A clinically significant history of poor response to bradykinin receptor 2 (BR2) blocker, C1- INH therapy or plasma kallikrein inhibitor therapy for the management of HAE, in the opinion of the Investigator.

[0675] 3. Use of angiotensin-converting enzyme (ACE) inhibitors after the Screening Visit or within 7 days prior to randomization.

[0676] 4. Any estrogen containing medications with systemic absorption (such as oral contraceptives including ethinylestradiol or hormonal replacement therapy) within 7 days prior to the Screening Visit.

[0677] 5. Use of strong cytochrome P450 3A4 CYP3A4 inhibitors and inducers during participation in the trial, starting within 5 half-lives of the Screening Visit.

[0678] Note: These medications include but are not limited to the following: Inhibitors: boceprevir, clarithromycin, cobicistat, dasabuvir, denoprevir, elvitegravir, idelalisib, indinavir, itraconazole, ketoconazole, lopinavir, nefazodone, nelfinavirombitasvir, paritaprevir, posaconazole, ritonavir, saquinavir, telaprevir, telithromycin, tipranavir, troleandomycin, and voriconazole.

[0679] Inducers: apalutamide, carbamazepine, enzalutamide, mitotane, phenytoin, rifampin, St. John’s Wort.

[0680] 6. Inadequate organ function, including but not limited to: a. Alanine aminotransferase (ALT) >2x upper limit of normal (ULN) b. Aspartate aminotransferase (AST) >2x ULN c. Bilirubin direct >1.25x ULN d. International normalized ratio (INR) >1.2 e. Clinically significant hepatic impairment defined as a Child-Pugh B or C.

[0681] 7. Any clinically significant comorbidity or systemic dysfunction, which in the opinion of the Investigator, would jeopardize the safety of the patient by participating in the trial.

[0682] 8. History of substance abuse or dependence that would interfere with the completion of the trial, as determined by the Investigator.

[0683] 9. Known hypersensitivity to the compound or placebo or to any of the excipients.

[0684] 10. Prior participation in the phase 2 trial.

[0685] 11. Participation in any gene therapy treatment or trial for HAE.

[0686] 12. Participation in any interventional investigational clinical trial, including an investigational COVID 19 vaccine trial, within 4 weeks of the last dosing of investigational drug prior to screening.

[0687] 13. Any pregnant or breastfeeding patient.

[0688] Assessments:

[0689] Efficacy Variables:

[0690] • Patient global impression of change (PGI-C) scored on a 7-point rating scale as much better, better, a little better, no change, a little worse, worse, much worse.

[0691] • Patient global impression of severity (PGI-S) scored on a 5-point rating scale as none, mild, moderate, severe, and very severe.

[0692] • Visual analog scale (VAS) anchored at 0 (none) and 100 (very severe) for abdominal pain, skin pain, and skin swelling.

[0693] • Modified General Anxiety - Numeric Rating Scale (GA-NRS) scored on an 11-point scale anchored at 0 (not at all anxious) and 10 (extremely anxious).

[0694] • Use of conventional on-demand treatment. Safety Variables:

[0695] • Adverse events, including serious adverse events.

[0696] • Laboratory test results.

[0697] • 12-lead ECG.

[0698] • Vital signs.

[0699] • Physical examination findings.

[0700] Criteria for Evaluation of Efficacy

[0701] Primary Efficacy Endpoints:

[0702] • PGI-C: Time to beginning of symptom relief defined as at least “a little better” (2 time points in a row) within 12 hours of the first study drug administration.

[0703] Key Secondary Efficacy Endpoints:

[0704] • PGI-S: Time to first incidence of decrease from baseline within 12 hours of the first study drug administration.

[0705] • PGI-S: Time to HAE attack resolution defined as “none” within 24 hours of the first study drug administration.

[0706] Secondary Efficacy Endpoints:

[0707] • PGI-C: Proportion of attacks with beginning of symptom relief defined as at least “a little better” (2 time points in a row) within 4 hours and within 12 hours of the first study drug administration.

[0708] • PGI-C: Time to at least “better” within 12 hours of the first study drug administration.

[0709] • PGI-S: Time to first incidence of decrease from baseline within 24 hours of the first study drug administration.

[0710] • Composite VAS: Time to at least a 50% decrease from baseline (3 time points in a row) within 12 hours and within 24 hours of the first study drug administration.

[0711] Exploratory Endpoints:

[0712] • GA-NRS: Cumulative GA-NRS expressed as area under the curve over 12 and 24 hours of the first study drug administration. General Statistical Methods and Types of Analyses

[0713] Analysis Sets:

[0714] • Safety Set will include all patients who receive at least one dose of trial medication.

[0715] • Full Analysis Set (FAS) will include all randomized patients who receive trial medication from at least two periods (including placebo period) after respective qualifying HAE attack. If one or more patient(s) received incorrect trial medication, data summarized using the FAS will be presented according to the randomized treatment. The FAS population will be the population for confirmatory efficacy analyses.

[0716] • Per-protocol Set (PPS) includes all patients from FAS who receive all three trial medications, complete scheduled assessments, and who do not have pre-defined protocol deviations that may affect primary efficacy endpoint.

[0717] Subgroup analysis sets

[0718] • On-demand Full Analysis Set on-demand FAS) will include FAS patients who enter the trial taking only conventional on-demand treatment.

[0719] • Prophylaxis Full Analysis Set (prophylaxis-FAS) will include FAS patients who enter the trial on a stable dose and regimen of long-term prophylactic treatment.

[0720] Sample size:

[0721] Approximately 114 patients will be randomized into the trial to ensure approximately 84 patients (including a minimum of 12 adolescents) complete the trial. The trial population will comprise 2 subsets: (1) patients who enter the trial taking only conventional on-demand treatment and (2) patients who enter the trial on a stable dose and regimen of long-term prophylactic treatment

[0722] 1. A sample size of 66 patients completing the trial would provide 90% power for testing each pairwise comparison (the compound versus placebo) at the 2.5% alpha level (2- sided) for the primary endpoint of time to beginning of symptom relief of the HAE attack as defined by PGI-C as at least “a little better” for 2 time points in a row within 12 hours of the first study drug administration. This sample size is derived based upon an assumption that median time to symptom relief of the HAE attack is 1.6 hours in active dose arm and 9 hours in placebo arm from the Phase 2 trial. It is assumed that patients will begin the trial together and will be followed for the same period of time, 49% of patients in control group and 17% of patients in the compound dose group are assumed to be lost-to-follow-up (right-censored). Taking a conservative approach and using simulation-based procedure for power calculations for a parallel group design, the two-sided two-group survival comparison Gehan Wilcoxon test has approximately 90% power to detect a median time ratio of 5.6 (9 / 1.6) with a target 2-sided significance level of 2.5% when there are 84 patients in each treatment group.

[0723] 2. The treatment effect observed in the Phase 2 trial (Example 15) is assumed to be representative of the entire population in this study. However, additional trial populations will be enrolled (eg, adolescent patients and patients that enter the trial on a stable dose and regimen of long-term prophylactic treatment where the treatment effect has not been previously characterized. Therefore a total of 84 patients completing the trial is proposed. This conservative approach increases the likelihood of maintaining at least 90% power in the event that the true treatment effect in this study population is different to that observed in the phase 2 trial.

[0724] Assuming there is approximately 30% dropout or non-completion rate, consistent with the Phase 2 trial, the oversampling by 30 patients (84 + 30 = 114) is proposed to account for patients that may not complete all treatment periods due to infrequent or ineligible HAE attacks or for patients who discontinue the trial early, for whatever reason.

[0725] General Considerations:

[0726] Continuous data will be summarized by treatment group using descriptive statistics (number, arithmetic mean, median, standard deviation, minimum, and maximum). Categorical data will be summarized by treatment group using frequency tables (frequencies and percentages).

[0727] All analyses will be carried out using SAS Version 9.4 or higher or using other validated software.

[0728] Sample Analyses:

[0729] The primary efficacy endpoint confirmatory analysis will have Bonferroni multiplicity adjustment for multiple dose levels, therefore pairwise comparison tests will be 2-sided with an alpha of 0.025. The analysis of the secondary or exploratory endpoints will not have multiplicity adjustments.

[0730] There are 3 treatment groups in the trial:

[0731] 1. 300 mg of the compound 2. 600 mg of the compound

[0732] 3. Placebo

[0733] Two pairwise comparisons will be performed: 300 mg of the compound versus placebo and 600 mg of the compound versus placebo.

[0734] Confirmatory efficacy analysis will be performed on the Full Analysis Set.

[0735] Efficacy Analyses:

[0736] A fixed sequence closed testing procedure will be followed. In a fixed sequence closed testing procedure the formal inferential testing can proceed to the next step only when statistical significance is declared in the current step. If the testing sequence is stopped, the remaining endpoints in the testing sequence will be considered exploratory. The fixed testing procedure will be employed first on the primary and then on the key secondary endpoints 1 and 2, separately for each dose comparison to placebo.

[0737] Statistical tests on both the primary and the key secondary endpoints will be at the same significance level alpha (0.025), Key secondary endpoint 1 will be tested only if the test on the primary endpoint is statistically significant. Testing within a dose level will be stopped if the test on the primary endpoint could not reject null hypothesis for the compound dose level at significance level of 0.025.

[0738] Key secondary endpoint 2 will be tested only if the test on the key secondary endpoint 1 is statistically significant. Testing within a dose level will be stopped if the test on the key secondary endpoint 1 could not reject null hypothesis for the compound dose level at significance level of 0.025.

[0739] Significance level 0.025 is Bonferroni adjusted significance level obtained by dividing the original significance level 0.05 by the number of comparisons within endpoint family between each compound dose level and placebo, i.e. the adjusted significance level is 0.025 (0.05 divided by 2).

[0740] The null hypothesis is that there is no difference in survival distribution of the time to beginning of symptom relief defined by the PGI-C as at least “a little better” for 2 time points in a row within 12 hours of the first study drug administration (no difference between each dose of the compound group versus placebo group) versus the alternative hypothesis that the survival distributions are different (each of the compound dose groups versus placebo). where tk is the time to “a little better” or higher rating of HAE attack following the compound dose treatment and tpis the time to “a little better” or higher rating of HAE attack following placebo.

[0741] WaU.: tkA. — [J =£ 0.

[0742] The primary endpoint will be analysed using Gehan Score Transformation test proposed by Feingold and Gillespie (1996) (Feingold M, Gillespie BW. Cross-over trials with censored data. Stat Med. 1996; 15:953-67) for crossover trials with censored data.

[0743] The number of patients with the primary endpoint event will be summarized by frequencies and survival estimates, the summaries will be presented by treatment.

[0744] HAE attack severity will be assessed on the PGI-S 5-point Likert scale scored as none, mild, moderate, severe, and very severe. A decrease in severity is defined as any change to any less severe level post baseline, than the score reported at baseline.

[0745] Key secondary endpoints will be analyzed with the same approach as primary endpoint analysis method (Gehan Score Transformation test). Key secondary endpoints will be tested according to the fixed sequence closed testing procedure. Key secondary endpoints will be summarized by frequencies and survival estimates; the summaries will be presented by treatment.

[0746] Subgroup analyses of the primary and key secondary efficacy endpoint will be performed by primary attack location at HAE attack onset, gender, age, Baseline severity, region, and number of doses received. Frequencies and survival estimates will be presented for each subgroup. Subgroups by time of attacks may also be investigated.

[0747] Analysis of the primary and key secondary efficacy endpoints will be performed on the FAS and the prophylaxis FAS. Secondary endpoints will be summarized by frequencies and percentages and presented by treatment.

[0748] All time-to-event endpoints will be summarized by frequencies of the events and Kaplan-Meier estimates or survival estimates as appropriate. The summaries will be presented by treatment.

[0749] For the exploratory endpoints baseline- and time-adjusted ALICs of the GA-NRS over time from 0 hour (pre-dose) up to 12 hours and up to 24 hours or up to the last time point prior to the time of conventional on-demand treatment use, whichever occurs first will be derived by linear trapezoidal rule. ALICs will be summarized descriptively. A statistical comparison of the ALICs between treatments will be performed using a mixed effects analysis of variance with fixed effects of treatment, sequence, and period and patient nested within sequence as a random effect.

[0750] Safety

[0751] Safety analyses will be performed by treatment group using the safety set. Safety endpoints include AE, clinical laboratory assessments, vital signs, and ECG findings.

[0752] Adverse events and serious adverse events (SAEs) recorded during the trial will be summarized by system organ class, preferred term, and treatment. Adverse events and medical history will be coded using the most current version of MedDRA.

[0753] Frequencies and percentages of patients with treatment-emergent adverse events (TEAEs), serious TEAEs, and TEAEs causing premature discontinuation will be provided by treatment group.

[0754] Patient listings of all Aes will be provided as well as listings of deaths, SAEs, and Aes leading to discontinuation.

[0755] For physical examination, ECG, vital signs, and laboratory variables (measured by the central laboratory), the number and percentage of patients with normal or abnormal results will be presented at each scheduled visit by sequence. Descriptive statistics for continuous variables will be provided at scheduled visits together with a summary of changes from baseline for each parameter by sequence.

[0756] Example 18 - Phase 1 multiple dose study in healthy adult subjects Aim: To evaluate the safety, tolerability, pharmacokinetics, and the change from baseline in QTc following administration of the compound formulated as 100 mg film coated tablets in healthy adult subjects.

[0757] Primary Objective:

[0758] • To investigate the safety and tolerability of multiple doses of the compound.

[0759] Secondary Objectives:

[0760] • To investigate the pharmacokinetics (PK) of multiple doses of the compound.

[0761] • To evaluate the effects of the compound on ECG parameters, including concentration-QTc relationship, following administration of the compound 100 mg Film Coated Tablets (Kai Vista Pharmaceuticals) to healthy adult subjects.

[0762] Exploratory Objectives:

[0763] • To investigate the pharmacodynamics (PD) of multiple doses of the compound.

[0764] Methods:

[0765] This was a phase 1 , double-blind, placebo-controlled, multiple-dose, multiple-cohort study to evaluate safety and tolerability of the compound as well as of the ECG effects of the compound formulated as 100 mg Film Coated Tablets in healthy adult male and female subjects.

[0766] Four (4) cohorts were evaluated. Cohorts 1 , 2 and 3 included 8 subjects each. Cohort 4 included 18 subjects. Every attempt was made to include an equal number of male and female subjects in each cohort.

[0767] During the study, oral doses of 600 mg of the compound as Film Coated Tablets (six 100 mg tablets) or 6 matching placebo tablets were administered once every 8 hours (Cohort 1) every 4 hours (Cohort 2), or every 2 hours (Cohort 3 and 4) to healthy adult male and female subjects up to a total dose of 1800 mg. In Cohorts 1 , 2 and 3, 6 subjects received the compound as 100 mg Film Coated Tablets and 2 subjects received the placebo for a total of 8 subjects per cohort. In Cohort 4, 12 subjects received the compound as 100 mg Film Coated Tablets and 6 subjects received the placebo for a total of 18 subjects.

[0768] Progression from Cohort 1 to Cohort 2 and Cohort 2 to Cohort 3 occurred after review of the safety data (labs, vital signs, safety ECGs, and adverse events) captured during the conduct of Cohort 1 and Cohort 2. Progression to Cohort 4 occurred after review of the safety data and pharmacokinetic data from Cohort 3. The pharmacokinetic data from Cohort 3 was reviewed to ensure that the Cmax of the 3rd dose is high enough to support the evaluation of the change in the QTc interval from baseline.

[0769] A Holter monitor was attached to each subject in order to continuously record ECGs. The monitor was attached 1 hour before the first dose and remained attached until after the final blood sample collection. The electrodes for the Holter monitor were checked by a member of the clinic staff at appropriate intervals to ensure they were attached.

[0770] Blood samples were collected at pre-dose, at intervals after the first dose, and at intervals over 24 hours after the final (third) dose (40 hours from the initial dose in Cohort 1 , 32 hours from the initial dose in Cohort 2, 28 hours from the initial dose in Cohorts 3 and 4) in each cohort. Subjects were confined to the clinical facility from at least 10 hours before dosing until after the final blood sample collection in each study cohort and returned to the clinic 5 to 7 days after the final dose for safety evaluations.

[0771] The pharmacokinetics of the compound were measured by a fully validated analytical procedure and the pharmacodynamic effect on plasma kallikrein inhibition enzyme activity was evaluated by an exploratory pharmacodynamic assessment.

[0772] Statistical analysis was performed to evaluate the relationship between plasma drug concentrations and the change from baseline in ECG effects of the test formulation.

[0773] Treatment administration

[0774] Cohort 1

[0775] The subjects received the test or placebo treatment every 8 hours over a 16-hour period (3 administrations of: 6 x 100 mg of the compound as 100 mg Film Coated Tablets or placebo dose administrations at O, 8, and 16 hours, total dose of 1800 mg of the compound or placebo) according to a two-treatment randomization schedule under direct observation. Each dose was administered with 240 mL of room temperature water. Subjects were instructed to swallow the tablets whole without chewing or biting. Any subject who bit or chewed the tablets was dropped from the study. Immediately after dosing a mouth check was performed

[0776] Cohort 2 The subjects received the test or placebo treatment every 4 hours over an 8-hour period (3 administrations of: 6 x 100 mg of the compound as 100 mg Film Coated Tablets or placebo dose administrations at 0, 4, and 8 hours, total dose of 1800 mg of the compound or placebo) according to a two-treatment randomization schedule under direct observation. Each dose was administered with 240 mL of room temperature water. Subjects were instructed to swallow the tablets whole without chewing or biting. Any subject who bit or chewed the tablets was dropped from the study. Immediately after dosing a mouth check was performed to ensure that the tablets were swallowed whole without chewing or biting.

[0777] Cohort 3 and 4

[0778] The subjects received the test or placebo treatment every 2 hours over a 4- hour period (3 administrations of: 6 x 100 mg of the compound as 100 mg Film Coated Tablets or placebo dose administrations at 0, 2, and 4 hours, total dose of 1800 mg of the compound or placebo) according to a two-treatment randomization schedule under direct observation. Each dose was administered with 240 mL of room temperature water. Subjects were instructed to swallow the tablets whole without chewing or biting. Any subject who bit or chewed the tablets were dropped from the study. Immediately after dosing a mouth check was performed to ensure that the tablets were swallowed whole without chewing or biting.

[0779] All subjects fasted (except water) for at least 8 hours before the first dosing. After initial dosing, subjects continued to fast until at least 6 hours after the first dose.

[0780] Method of Assigning Subjects to Treatment Groups:

[0781] Cohort 1, 2 and 3

[0782] Subjects were randomized such that 6 subjects received the test product and 2 subjects received the placebo. As a safety measure, a sentinel dosing scheme was incorporated for each cohort, in which one subject will receive the test product and one subject received the placebo product followed by the remainder of the cohort.

[0783] Cohort 4

[0784] Subjects were randomized such that 12 subjects received the test product and 6 subjects received the placebo.

[0785] The randomization schedule was generated prior to the first dosing cohort using SAS®, Version 9.4 or higher. Results:

[0786] No serious adverse events were reported during the study and no subjects were discontinued because of an AE. All reported adverse events were considered “mild” in severity and had an outcome of “recovered / resolved” at the end of the study.

[0787] No clinically relevant effects on the studied ECG parameters were identified.

[0788] Figure 29a shows the mean plasma concentrations of the compound of Formula A after the initial dose for each cohort.

[0789] Figure 29b shows the mean plasma concentrations (semi-logarithmic scale) of the compound for formula A for each cohort.

[0790] These data demonstrate that the compound of Formula A has a pharmacokinetic profile suitable for oral administration when administered in multiple dosage amounts. The results further suggest that the compound of Formula A can be dosed safely at regular intervals.

[0791] Also disclosed are the following numbered embodiments:

[0792] 1. A solid form of the compound of Formula A: which exhibits at least the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately 5.8, 10.1 , 15.0, 16.1 and 17.3.

[0793] 2. The solid form according to embodiment 1 having an X-ray powder diffraction pattern substantially the same as that shown in Figure 12. The solid form according to embodiment 1 or 2, which exhibits an endothermic peak in its DSC thermograph at 163 ± 3°C. The solid form according to any one of embodiments 1 to 3 having a DSC thermograph substantially the same as that shown in Figure 13. A solid form of the compound of Formula A: which exhibits an endothermic peak in its DSC thermograph at 163 ± 3°C. The solid form according to embodiment 5 having a DSC thermograph substantially the same as that shown in Figure 13. A pharmaceutical composition comprising the solid form according to any one of embodiments 1 to 6, and a pharmaceutically acceptable carrier, diluent or excipient. A solid form according to any one of embodiments 1 to 6, for use in therapy. A solid form according to any one of embodiments 1 to 6, for use in the treatment of a disease or condition mediated by plasma kallikrein. A method of treatment of a disease or condition mediated by plasma kallikrein, said method comprising administering to a mammal in need of such treatment a therapeutically effective amount of a solid form according to any one of embodiments 1 to 6. The solid form for use according to embodiment 9, or the method according to embodiment 10, wherein the disease or condition mediated by plasma kallikrein is selected from impaired visual acuity, diabetic retinopathy, retinal vascular permeability associated with diabetic retinopathy, diabetic macular edema, hereditary angioedema, diabetes, pancreatitis, cerebral haemorrhage, nephropathy, cardiomyopathy, neuropathy, inflammatory bowel disease, arthritis, inflammation, septic shock, hypotension, cancer, adult respiratory distress syndrome, disseminated intravascular coagulation, blood coagulation during cardiopulmonary bypass surgery and bleeding from post-operative surgery. The solid form for use according to embodiment 9, or the method according to embodiment 10, wherein the disease or condition mediated by plasma kallikrein is selected from retinal vascular permeability associated with diabetic retinopathy, diabetic macular edema and hereditary angioedema. The solid form for use according to embodiment 12, or the method according to embodiment 12, wherein the disease or condition mediated by plasma kallikrein is selected from retinal vascular permeability associated with diabetic retinopathy, and diabetic macular edema. The solid form for use according to embodiment 12, or the method according to embodiment 12, wherein the disease or condition mediated by plasma kallikrein is hereditary angioedema. The solid form for use according to embodiment 12, or the method according to embodiment 12, wherein the disease or condition mediated by plasma kallikrein is diabetic macular edema. The solid form for use according to embodiment 9, or the method according to embodiment 10, wherein the disease or condition mediated by plasma kallikrein is retinal vein occlusion. The solid form for use according to embodiment 13 or embodiment 15, or the method according to embodiment 13 or embodiment 15, wherein said solid form is administered in a form suitable for injection into the ocular region of a patient, in particular, in a form suitable for intra-vitreal injection. A process for the preparation of a solid form according to any one of embodiments 1 to 6, comprising crystallising said solid form from a mixture of the compound of the Formula A and a solvent or a mixture of solvents. The process of embodiment 18, wherein the solvent is isopropanol. The process of embodiment 18 or 19, wherein said mixture is heated to an elevated temperature. The process of embodiment 20, wherein the elevated temperature is about 78-83°C, for example 78-82°C. The process of embodiment 20 or 21, wherein, after heating, said mixture is cooled. The process of embodiment 22, wherein the mixture is cooled to a temperature of about -80°C to about 25°C, preferably about -20°C to about 25°C, more preferably about 0°C to about 25°C, most preferably about 0°C to about 5°C. A screening method for determining whether the solid form of the compound of Formula A according to any one of embodiments 1 to 6 (Form 19) is present in a sample comprising any form of the compound of Formula A, wherein the screening method comprises the steps of: a. Providing the sample; b. Measuring a property of the sample; and c. Determining whether the solid form of the compound of Formula A according to any one of embodiments 1 to 6 is present in the sample by comparing the result of step b with one or more known results of measurements of the same property for one or more known solid forms of the compound of Formula A. The screening method of embodiment 24, wherein the measured property of the sample is compared with known values of a measurement of the same property for the solid form of the compound of Formula A according to any one of embodiments 1 to 6. The screening method of embodiment 24 or 25, wherein the sample comprises, or consists essentially of, a solid form of the compound of Formula A (Form 1) which: a. exhibits at least the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately 11.2, 12.5, 13.2, 14.5 and 16.3; and / or b. has an X-ray powder diffraction pattern substantially the same as that shown in Figure 2a; and / or c. exhibits an endothermic peak in its DSC thermograph at about 151 °C. The screening method of any one of embodiments 24 to 26, wherein the measured property of the sample is compared with known values of a measurement of the same property for a solid form of the compound of Formula A (Form 1) which: a. exhibits at least the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately 11.2, 12.5, 13.2, 14.5 and 16.3; and / or b. has an X-ray powder diffraction pattern substantially the same as that shown in Figure 2a; and / or c. exhibits an endothermic peak in its DSC thermograph at about 151 °C. The screening method of any one of embodiments 24 to 27, wherein measuring a property of the sample comprises performing one or more of X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), differential thermal analysis (DTA), thermogravimetric analysis (TGA), gravimetric vapour sorption (GVS), simultaneous thermal analysis (STA), infrared (IR) spectroscopy on the sample. The screening method of embodiment 28, wherein measuring a property of the sample comprises performing differential scanning calorimetry (DSC) on the sample. The screening method of embodiment 29, wherein a DSC thermograph is obtained. The screening method of embodiment 29, wherein the result of the measurement of the property of the sample is a temperature value of an endothermic peak in a DSC thermograph, preferably an endothermic peak corresponding to a melting temperature of a solid form of the compound of Formula A. The screening method of embodiment 30, wherein the result of step b comprises an onset temperature of an endothermic melting peak in the DSC thermograph. The screening method of embodiment 31 , wherein the one or more known results include an onset temperature for an endothermic melting peak in a DSC thermograph obtained for the solid form of the compound of Formula A according to any one of embodiments 1 to 6. The screening method of embodiment 32, wherein the one or more known results include an onset temperature for an endothermic melting peak in a DSC thermograph at about 158°C. The screening method of embodiment 33, wherein it is determined that Form 19 is present in the sample if an endothermic melting peak is present in a DSC thermograph obtained for the sample with an onset temperature between 156°C and 161 °C, optionally at about 158°C.

[0794] Ill The screening method of embodiment 24, wherein the screening method comprises the steps of: a. Providing the sample; b. Measuring a property of the sample, comprising performing DSC on the sample to obtain a DSC thermograph; and c. Determining whether Form 19 is present in the sample by comparing the DSC thermograph of step b with a DSC thermograph obtained by performing DSC on Form 1 and / or Form 19, preferably Form 19. The screening method of any one of embodiments 24 to 35, wherein, if it is determined that Form 19 is present in the sample, the method further comprises the steps of: d. Providing a reference sample comprising Form 19; e. Measuring the same property of the reference sample; g. Determining whether the sample comprises an amount of Form 19 which is less than, equal to, or greater than the amount Form 19 present in the reference sample by comparing the result of step b with the result of step e. The screening method of embodiment 37, wherein step b comprises performing DSC on the sample to obtain a DSC thermograph and step e comprises performing DSC on the reference sample to obtain a DSC thermograph. The screening method of embodiment 37 or 38, wherein the sample consists essentially of Form 1 and Form 19; wherein Form 1 is the solid form of the compound of Formula A (Form 1) which: a. exhibits at least the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately 11.2, 12.5, 13.2, 14.5 and 16.3; and / or b. has an X-ray powder diffraction pattern substantially the same as that shown in Figure 2a; and / or c. exhibits an endothermic peak in its DSC thermograph at about 151 °C. The screening method of any one of embodiments 37 to 39, wherein the reference sample consists essentially of Form 1 and Form 19; wherein Form 1 is the solid form of the compound of Formula A (Form 1) which: a. exhibits at least the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately 11.2, 12.5, 13.2, 14.5 and 16.3; and / or b. has an X-ray powder diffraction pattern substantially the same as that shown in Figure 2a; and / or c. exhibits an endothermic peak in its DSC thermograph at about 151 °C. The screening method of embodiment 40, wherein the reference sample consists essentially of Form 1 and about 0.5 wt.% of Form 19. The screening method of any one of embodiments 37 to 41, wherein step b comprises performing DSC on the sample multiple times to obtain multiple DSC thermographs. The screening method of any one of embodiments 37 to 42, wherein step g comprises comparing one or more DSC thermographs of step b with the DSC thermograph of step e. The screening method of embodiment 43, wherein comparing the one or more DSC thermographs of step b with the DSC thermograph of step e comprises comparing values determined from the endothermic melting peaks in the one or more DSC thermographs of step b with values determined from the endothermic melting peaks in the DSC thermograph of step e. The screening method of embodiment 44, wherein the endothermic melting peaks in the one or more DSC thermographs of step b and the endothermic melting peaks in the one or more DSC thermographs of step e are the endothermic melting peaks corresponding to Form 1 and Form 19. The screening method of embodiment 45, wherein the endothermic melting peak in a DSC thermograph corresponding to Form 1 is a peak with an onset temperature in the range of 150°C and 155°C. The screening method of embodiment 45, wherein the endothermic melting peak in a DSC thermograph corresponding to Form 1 is a peak with an onset temperature at about 151°C. The screening method of any one of embodiments 45 to 47, wherein the endothermic melting peak in a DSC thermograph corresponding to Form 19 is a peak with an onset temperature in the range of 156°C and 161°C. The screening method of any one of embodiments 45 to 47, wherein the endothermic melting peak in a DSC thermograph corresponding to Form 19 is a peak with an onset temperature at about 158°C. The screening method of any one of embodiments 44 to 49, wherein the values determined from the endothermic melting peaks in the one or more DSC thermographs of step b and the values determined from the endothermic melting peaks in the DSC thermograph of step e are the peak areas. The screening method of any one of embodiments 45 to 49, wherein the values determined from the endothermic melting peaks in the one or more DSC thermographs of step b and the values determined from the endothermic melting peaks in the DSC thermograph of step e are the melting enthalpies of the Form 1 and Form 19 present in the sample and reference sample. The screening method of embodiment 51 , wherein determining whether the sample comprises an amount of Form 19 which is less than, equal to, or greater than the amount of Form 19 present in the reference sample comprises comparing the melting enthalpies calculated from the endothermic melting peaks in the DSC thermographs by calculating the ratio of the melting enthalpies for the Form 1 and Form 19 content of the sample (Rt) and the reference sample (Rr). The screening method of embodiment 52, wherein the melting enthalpies are normalised melting enthalpies.

Claims

CLAIMS1. A solid form of the compound of Formula A:which exhibits at least the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately 5.8, 10.1 , 15.0, 16.1 and 17.3.

2. The solid form according to claim 1 having an X-ray powder diffraction pattern substantially the same as that shown in Figure 12.

3. The solid form according to claim 1 or 2, which exhibits an endothermic peak in its DSC thermograph at 163 ± 3°C.

4. The solid form according to any one of claims 1 to 3 having a DSC thermograph substantially the same as that shown in Figure 13.

5. A solid form of the compound of Formula A:which exhibits an endothermic peak in its DSC thermograph at 163 ± 3°C.

6. The solid form according to claim 5 having a DSC thermograph substantially the same as that shown in Figure 13.

7. A pharmaceutical composition comprising a solid form as claimed in any one of claims 1 to 6, and a pharmaceutically acceptable carrier, diluent or excipient.

8. A process for the preparation of a solid form as claimed in any one of claims 1 to 6, comprising crystallising said solid form from a mixture of the compound of the Formula A and a solvent or a mixture of solvents.

9. The process of claim 8, wherein the solvent is isopropanol.

10. The process of claim 8 or 9, wherein said mixture is heated to an elevated temperature; optionally wherein the elevated temperature is about 78-83°C, for example 78-82°C.

11. The process of claim 10, wherein, after heating, said mixture is cooled; optionally wherein the mixture is cooled to a temperature of about -80°C to about 25°C, preferably about -20°C to about 25°C, more preferably about 0°C to about 25°C, most preferably about 0°C to about 5°C.

12. A screening method for determining whether the solid form of the compound of Formula A according to any one of claims 1 to 6 (Form 19) is present in a sample comprising any form of the compound of Formula A, wherein the screening method comprises the steps of: a. Providing the sample; b. Measuring a property of the sample; and c. Determining whether the solid form of the compound of Formula A according to any one of claims 1 to 6 is present in the sample by comparing the result of step b with one or more known results of measurements of the same property for one or more known solid forms of the compound of Formula A.

13. The screening method of claim 12, wherein the measured property of the sample is compared with known values of a measurement of the same property for the solid form of the compound of Formula A according to any one of claims 1 to 6.

14. The screening method of claim 12 or 13, wherein the sample comprises, or consists essentially of, a solid form of the compound of Formula A (Form 1) which:a. exhibits at least the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately 11.2, 12.5, 13.2, 14.5 and 16.3; and / or b. has an X-ray powder diffraction pattern substantially the same as that shown in Figure 2a; and / or c. exhibits an endothermic peak in its DSC thermograph at about 151 °C.

15. The screening method of any one of claims 12 to 14, wherein the measured property of the sample is compared with known values of a measurement of the same property for a solid form of the compound of Formula A (Form 1) which: a. exhibits at least the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately 11.2, 12.5, 13.2, 14.5 and 16.3; and / or b. has an X-ray powder diffraction pattern substantially the same as that shown in Figure 2a; and / or c. exhibits an endothermic peak in its DSC thermograph at about 151 °C.

16. The screening method of any one of claims 12 to 15, wherein measuring a property of the sample comprises performing one or more of X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), differential thermal analysis (DTA), thermogravimetric analysis (TGA), gravimetric vapour sorption (GVS), simultaneous thermal analysis (STA), infrared (IR) spectroscopy on the sample; optionally wherein measuring a property of the sample comprises performing differential scanning calorimetry (DSC) on the sample.

17. The screening method of claim 16, wherein a DSC thermograph is obtained.

18. The screening method of claim 16, wherein the result of the measurement of the property of the sample is a temperature value of an endothermic peak in a DSC thermograph, preferably an endothermic peak corresponding to a melting temperature of a solid form of the compound of Formula A.

19. The screening method of claim 17, wherein the result of step b comprises an onset temperature of an endothermic melting peak in the DSC thermograph.

20. The screening method of claim 18, wherein the one or more known results include an onset temperature for an endothermic melting peak in a DSC thermograph obtained for the solid form of the compound of Formula A according to any one of claims 1 to 6; optionally wherein the one or more known results include an onset temperature for an endothermic melting peak in a DSC thermograph at about 158°C.

21. The screening method of claim 20, wherein it is determined that Form 19 is present in the sample if an endothermic melting peak is present in a DSC thermograph obtained for the sample with an onset temperature between 156°C and 161°C, optionally at about 158°C.

22. The screening method of claim 12, wherein the screening method comprises the steps of: a. Providing the sample; b. Measuring a property of the sample, comprising performing DSC on the sample to obtain a DSC thermograph; and c. Determining whether Form 19 is present in the sample by comparing the DSC thermograph of step b with a DSC thermograph obtained by performing DSC on Form 1 and / or Form 19, preferably Form 19.

23. The screening method of any one of claims 12 to 21 , wherein, if it is determined that Form 19 is present in the sample, the method further comprises the steps of: d. Providing a reference sample comprising Form 19; e. Measuring the same property of the reference sample; h. Determining whether the sample comprises an amount of Form 19 which is less than, equal to, or greater than the amount Form 19 present in the reference sample by comparing the result of step b with the result of step e.

24. The screening method of claim 23, wherein step b comprises performing DSC on the sample to obtain a DSC thermograph and step e comprises performing DSC on the reference sample to obtain a DSC thermograph.

25. The screening method of claim 23 or 24, wherein the sample consists essentially of Form 1 and Form 19; wherein Form 1 is the solid form of the compound of Formula A (Form 1) which: a. exhibits at least the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately 11.2, 12.5, 13.2, 14.5 and 16.3; and / orb. has an X-ray powder diffraction pattern substantially the same as that shown in Figure 2a; and / or c. exhibits an endothermic peak in its DSC thermograph at about 151 °C.

26. The screening method of any one of claims 23 to 25, wherein the reference sample consists essentially of Form 1 and Form 19; wherein Form 1 is the solid form of the compound of Formula A (Form 1) which: a. exhibits at least the following characteristic X-ray powder diffraction peaks (Cu Ka radiation, expressed in degrees 20) at approximately 11.2, 12.5, 13.2, 14.5 and 16.3; and / or b. has an X-ray powder diffraction pattern substantially the same as that shown in Figure 2a; and / or c. exhibits an endothermic peak in its DSC thermograph at about 151 °C.

27. The screening method of claim 26, wherein the reference sample consists essentially of Form 1 and about 0.5 wt.% of Form 19.

28. The screening method of any one of claims 23 to 27, wherein step g comprises comparing one or more DSC thermographs of step b with the DSC thermograph of step e.

29. The screening method of claim 28, wherein comparing the one or more DSC thermographs of step b with the DSC thermograph of step e comprises comparing values determined from the endothermic melting peaks in the one or more DSC thermographs of step b with values determined from the endothermic melting peaks in the DSC thermograph of step e.

30. The screening method of claim 29, wherein the endothermic melting peaks in the one or more DSC thermographs of step b and the endothermic melting peaks in the one or more DSC thermographs of step e are the endothermic melting peaks corresponding to Form 1 and Form 19.

31. The screening method of claim 30, wherein the endothermic melting peak in a DSC thermograph corresponding to Form 1 is a peak with an onset temperature in the range of 150°C and 155°C; optionally wherein the endothermic melting peak in a DSC thermograph corresponding to Form 1 is a peak with an onset temperature at about 151°C.

32. The screening method of claim 30 or 31 , wherein the endothermic melting peak in a DSC thermograph corresponding to Form 19 is a peak with an onset temperature in the range of 156°C and 161°C; optionally wherein the endothermic melting peak in a DSC thermograph corresponding to Form 19 is a peak with an onset temperature at about 158°C.

33. The screening method of any one of claims 30 to 32, wherein the values determined from the endothermic melting peaks in the one or more DSC thermographs of step b and the values determined from the endothermic melting peaks in the DSC thermograph of step e are the melting enthalpies of the Form 1 and Form 19 present in the sample and reference sample.

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