RU2817800C2 - Therapeutic compounds and compositions - Google Patents

Abstract

FIELD: pharmaceutics.

SUBSTANCE: invention relates to compounds of formula (II), where R¹–R⁵, R^x, R^a, R^b, m, n are defined in the patent claim, which inhibit factor XIa or kallikrein, and their pharmaceutically acceptable salts and pharmaceutical compositions. Also disclosed are methods of treating or reducing the risk of thromboembolic complications, stroke, acute coronary syndrome, thromboembolic disorder or atrial fibrillation by inhibiting factor XIa in a patient.

EFFECT: therapeutic compounds and compositions are disclosed.

28 cl, 9 tbl, 221 ex

Description

CROSS-REFERENCES TO RELATED APPLICATIONS

This application claims the benefit of provisional patent application US 62/627435, filed February 7, 2018, which is incorporated herein by reference in its entirety.

BACKGROUND OF THE ART

Blood coagulation represents the first line of defense against blood loss after injury. The blood coagulation “cascade” involves a number of circulating serine protease zymogens, regulatory cofactors, and inhibitors. Each enzyme, once produced from its zymogen, specifically cleaves the next zymogen in the cascade to produce an active protease. This process is repeated until finally thrombin breaks down fibrinopeptides from fibrinogen to produce fibrin, which polymerizes to form a blood clot. Although effective coagulation limits blood loss at the site of injury, it also creates the risk of systemic coagulation leading to massive thrombosis. Under normal conditions, hemostasis maintains a balance between clot formation (coagulation) and clot dissolution (fibrinolysis). However, in some pathological conditions, such as acute myocardial infarction and unstable angina, rupture of an established atherosclerotic plaque leads to abnormal thrombus formation in the vasculature of the coronary arteries.

Diseases caused by blood coagulation, such as myocardial infarction, unstable angina, atrial fibrillation, stroke, pulmonary embolism and deep vein thrombosis, are the leading causes of death in developed countries. Current treatments with anticoagulant therapeutic agents, such as unfractionated heparin injection, low molecular weight (LMW) heparin, and oral warfarin (Coumadin), can be dangerous due to the risk of bleeding episodes and have variable effects on patients, leading to the need for careful monitoring and selection. therapeutic doses. Consequently, today there is a need in medicine for new anticoagulant drugs that do not exhibit some or all of the side effects that currently available drugs have.

Factor XIa is an attractive therapeutic target involved in metabolism associated with these diseases. Elevated levels of factor XIa or increased activity of factor XIa have been observed in several thromboembolic diseases, including venous thrombosis (Meijers et al., N. Engl. J. Med. 342:696, 2000), acute myocardial infarction (Minnema et al., Arterioscler Thromb Vasc Biol 20:2489, 2000), acute coronary syndrome (Butenas et al., Thromb. Haemost. 99:142, 2008), coronary heart disease (Butenas et al., Thromb. Haemost. 99:142, 2008), chronic obstructive pulmonary disease (Jankowski et al., Thromb. Res. 127:242, 2011), aortic stenosis (Blood Coagul Fibrinolysis, 22:473, 2011), acute cerebrovascular insufficiency (Undas et al., Eur. J. Clin. Invest ., 42:123, 2012) and systolic heart failure due to ischemic cardiomyopathy (Zabcyk et al., Pol. Arch. Med. Wewn. 120:334, 2010). Patients who lack factor XI due to genetic factor XI deficiency suffer virtually no ischemic strokes (Salomon et al., Blood, 111:4113, 2008). At the same time, the loss of factor XIa activity, which leaves intact one of the metabolic pathways that initiates coagulation, does not impair hemostasis. In humans, factor XI deficiency can lead to mild to moderate bleeding disorders, particularly in tissues with high levels of local fibrinolytic activity such as the urinary tract, nose, mouth, and tonsils. In addition, hemostasis is almost normal in factor XI-deficient mice (Gailani, Blood Coagul Fibrinolysis, 8:134, 1997). Further, inhibition of factor XI has also been found to improve hypertension and other diseases and dysfunctions, including vascular inflammation (Kossmann et al. Sci. Transl. Med. 9, eaah4923 (2017)).

Therefore, compounds that inhibit factor XIa have the potential to prevent or treat a wide range of disorders without the side effects and therapeutic problems that plague drugs that inhibit other components of the coagulation cascade. In addition, due to the limited effectiveness and adverse side effects of some current therapeutic agents for inhibiting unwanted thrombosis (eg, deep vein thrombosis, hepatic vein thrombosis and stroke), more effective compounds and methods (eg, factor XIa-related compounds and methods) are needed. to prevent or treat unwanted thrombosis.

Another therapeutic target is the venodilator enzyme kallikrein. Human plasma kallikrein is a serine protease that may be responsible for the activation of several downstream factors (e.g., bradykinin and plasmin) critical for coagulation and control of, for example, blood pressure, inflammation, and pain. Kallikreins are expressed, for example, in the prostate gland, epidermis and central nervous system (CNS) and may be involved, for example, in the regulation of sperm liquefaction, breakdown of cell adhesion proteins and neuronal plasticity in the CNS. In addition, kallikreins may be involved in tumorigenesis and the development of cancer and angioedema, such as hereditary angioedema. Excessive activation of the kallinkrein-kinin pathway can lead to a number of disorders, including angioedema, such as hereditary angioedema (HAE) (Schneider et al., J. Allergy Clin. Immunol. 120:2, 416, 2007). To date, there are limited treatment options for HAE (see, for example, WO2003/076458). Thus, there is a need for therapeutic agents to prevent or treat these diseases.

SUMMARY OF THE INVENTION

The present invention relates to compounds that inhibit factor XIa or kallikrein, and methods of preventing or treating unwanted thrombosis or angioedema (eg, hereditary angioedema) by administering one or more compounds alone or in combination with other active substances. The invention also relates to methods for developing and selecting additional factor XIa or kallikrein inhibitors using these structures. It is desirable that these compounds contain specific structures with physical and spatial characteristics that allow the compounds to interact with specific active site residues of factor XIa or kallikrein.

In one aspect, the present invention provides a compound of formula (II):

or a pharmaceutically acceptable salt thereof, wherein R ¹ is hydrogen or -NR ⁸ R ⁹ ; R ^a is C _1-6 alkyl, C _1-6 haloalkyl, halogen, cyano or -OR ⁶ ; R ^b represents hydrogen or C _1-6 alkyl; R ² represents an optionally substituted 5-membered heteroaryl or an optionally substituted 5-membered heterocyclyl; R ³ represents hydrogen, C _1-6 alkyl or C _1-6 haloalkyl; R ⁴ is C _1-6 alkyl, C _1-6 haloalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted with one, two or three independent substituents selected from halogen, C _{1- 6} alkyl, C _1-6 haloalkyl, cyano or -OR ⁶ ; R ⁵ is hydrogen, C _1-6 alkyl, C _1-6 haloalkyl, cycloalkyl or aryl, wherein cycloalkyl or aryl is optionally substituted with one, two or three independent substituents selected from halogen, C _1-6 alkyl, C _{1- 6} haloalkyl, cyano or -OR ⁶ , or R ⁴ and R ⁵ together with the carbon atom to which they are attached form a ring; R ⁶ represents hydrogen, C _1-6 alkyl, or C _1-6 haloalkyl; R ⁸ and R ⁹ are each independently hydrogen, C _1-6 alkyl, -C(O)R ¹⁰ or -C(O)OR ¹⁰ ; R ¹⁰ represents C _1-6 alkyl or C _1-6 haloalkyl; R ^x is -O or absent, where when R ^x is -O, the nitrogen atom of the pyridyl ring is positively charged and R ^x is negatively charged, thereby forming a pyridyl-N-oxide; m is 0, 1, 2 or 3; and n is 0 or 1, where if n is 0, then R ⁵ is hydrogen and R ⁴ is absent.

In one aspect, the compound described in the present invention is a compound of formula (II):

or a pharmaceutically acceptable salt thereof, wherein R ¹ is hydrogen or -NR ⁸ R ⁹ ; R ^a is C _1-6 alkyl, C _1-6 haloalkyl, halogen, cyano or -OR ⁶ ; R ^b represents hydrogen or C _1-6 alkyl; R ² represents an optionally substituted 5-membered heteroaryl; R ³ represents hydrogen, C _1-6 alkyl or C _1-6 haloalkyl; R ⁴ is C _1-6 alkyl, C _1-6 haloalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted with one, two or three independent substituents selected from halogen, C _{1- 6} alkyl, C _1-6 haloalkyl, cyano or -OR ⁶ ; R ⁵ is hydrogen, C _1-6 alkyl, C _1-6 haloalkyl, cycloalkyl or aryl, wherein cycloalkyl or aryl is optionally substituted with one, two or three independent substituents selected from halogen, C _1-6 alkyl, C _{1- 6} haloalkyl, cyano or -OR ⁶ , or R ⁴ and R ⁵ together with the carbon atom to which they are attached form a ring; R ⁶ represents hydrogen, C _1-6 alkyl or C _1-6 haloalkyl; R ⁸ and R ⁹ are each independently hydrogen, C _1-6 alkyl, -C(O)R ¹⁰ or -C(O)OR ¹⁰ ; R ¹⁰ represents C _1-6 alkyl or C _1-6 haloalkyl; m is 0, 1, 2 or 3; and n is 0 or 1, where if n is 0, then R ⁵ is hydrogen and R ⁴ is absent.

In some embodiments, the compound is a compound of formula (I):

In some embodiments, R ⁶ is C _1-6 alkyl or C _1-6 haloalkyl. In some embodiments, R ⁶ is -CH ₃ or -CF ₃ .

In some embodiments, R ¹ is hydrogen, -NH ₂ or -NHCH ₃ .

In some embodiments, R ² is a 5-membered heteroaryl, optionally substituted with one or two independent substituents selected from C _1-6 alkyl. In some embodiments, R ² is pyrazolyl, imidazolyl, thienyl, isothiazolyl, or thiazolyl, wherein imidazolyl, thienyl, isothiazolyl, or thiazolyl is optionally substituted with one or two independent substituents selected from C _1-6 alkyl. In some embodiments, R ² is pyrazolyl, imidazolyl, thienyl, isothiazolyl, or thiazolyl, wherein imidazolyl, thienyl, isothiazolyl, or thiazolyl is optionally substituted with one or two substituents selected from -CH ₃ .

In some embodiments, R ² is

In some embodiments, R ³ is -CH ₃ .

In some embodiments, R ⁴ is C _1-6 alkyl, cycloalkyl, aryl or heteroaryl, wherein the cycloalkyl, aryl or heteroaryl is optionally substituted with one, two or three independent substituents selected from halogen or C _1-6 alkyl. In some embodiments, R ⁴ is cyclohexyl, cyclopropyl, phenyl, pyridyl, pyrazolyl, thienyl, or C _1-6 alkyl, wherein cyclohexyl, cyclopropyl, phenyl, pyridyl, pyrazolyl, or thienyl is optionally substituted with one, two, or three independent substituents selected from halogen or C _1-6 alkyl. In some embodiments, R ⁴ is cyclohexyl, phenyl, pyridyl, pyrazolyl, or thienyl, wherein the cyclohexyl, phenyl, pyridyl, pyrazolyl, or thienyl is optionally substituted with one, two, or three independent substituents selected from halogen or C _1-6 alkyl. In some embodiments, R ⁴ is phenyl, optionally substituted with one, two or three substituents selected from -F, -Cl or C _1-6 alkyl.

In some embodiments, R ⁵ is hydrogen, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH ₂ CH ₃ , -CH(CH ₃ ) ₂ , -CH ₂ CH ₂ CH ₂ CH ₃ , -CF ₃ or unsubstituted cyclopropyl. In some embodiments, R ⁵ is -CH ₂ CH ₃ , -CF ₃ or unsubstituted cyclopropyl.

In some embodiments, R ⁴ is C _1-6 alkyl, C _1-6 haloalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted with one, two or three independent substituents selected from halogen , C _1-6 alkyl, C _1-6 haloalkyl, cyano or -OR ⁶ , and R ⁵ is hydrogen, C _1-6 alkyl, C _1-6 haloalkyl, cycloalkyl or aryl, wherein cycloalkyl or aryl is optionally substituted with one , two or three independent substituents selected from halogen, C _1-6 alkyl, C _1-6 haloalkyl or -OR ⁶ ; or R ⁴ and R ⁵ together with the carbon atom to which they are attached form a ring.

In some embodiments, the compound is a compound of formula (I-a):

In some embodiments, the compound is a compound of formula (I-b):

In some embodiments, the compound is a compound of formula (I-c), formula (I-d), formula (I-e), formula (I-f), formula (I-g), formula (I-i), or formula (I-j):

In some embodiments, the compound is a compound of formula (I-c):

In some embodiments, the compound is a compound of formula (I-e), formula (I-k), formula (I-l), formula (I-m), formula (I-o) or formula (I-p):

where each of R ^b2 independently represents halogen, C _1-6 alkyl, C _1-6 haloalkyl, cyano or -OR ⁶ ; and p is 0, 1, 2, or 3. In some embodiments, R ^a is -C _1-6 alkyl.

In some embodiments, R ^a is -CH ₃ . In some embodiments, R ^a is -CH ₃ and m is 1. In some embodiments, m is 1. In some embodiments, R ^b is C _1-6 alkyl. In some embodiments, R ^b is -CH ₃ .

In some embodiments, the compound is a compound selected from the compounds presented in Table 1.

In some embodiments, the compound is a pharmaceutically acceptable salt (eg, hydrochloride (HCl), hydrobromide (HBr), tartrate, oleate, or citrate salt). In a preferred embodiment, the pharmaceutically acceptable salt is a hydrochloride (HCl) salt.

In one aspect, the present invention relates to a pharmaceutical composition comprising a compound of formula (II) or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. In some embodiments, the composition is in the form of a liquid preparation (eg, solution). In some embodiments, the composition is in the form of a solid preparation (eg, capsule, pill, tablet, or powder).

In one aspect, the present invention relates to a method of reducing the risk of stroke (eg, ischemia, in particular transient ischemic attack) in a patient who has suffered an ischemic attack (eg, transient ischemic attack), comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutical thereof an acceptable salt or composition of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, administration reduces the patient's risk of stroke compared to a patient not administered the compound. In some embodiments, administration reduces the risk of atrial fibrillation in a patient compared to a patient not administered the compound.

In one aspect, the present invention provides a method of reducing the risk of systemic embolism outside the CNS (eg, ischemia, particularly transient ischemic attack) in a patient who has suffered an ischemic attack (eg, transient ischemic attack), comprising administering to the patient an effective amount of a compound of formula (II ) or a pharmaceutically acceptable salt or composition thereof of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, administration reduces the risk of systemic embolism outside the CNS in a patient compared to a patient who is not administered the compound.

In one aspect, the present invention provides a method of treating deep vein thrombosis, comprising administering to a patient who has suffered an ischemic attack (eg, transient ischemic attack), an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention (eg, a composition comprising a compound of formula (II)).

In one aspect, the present invention relates to a method of treating acute coronary syndrome in a patient, comprising administering to a patient in need thereof a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising compound of formula (II).

In one aspect, the present invention relates to a method of preventing deep vein thrombosis, comprising administering to a patient who has suffered deep vein thrombosis (for example, a patient who has previously been treated for deep vein thrombosis), an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof according to the present invention (eg, a composition comprising a compound of formula (II)). In one aspect, the present invention provides a method of reducing the risk of recurrent deep vein thrombosis, comprising administering to a patient who has experienced deep vein thrombosis (for example, a patient who has previously been treated for deep vein thrombosis), an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof, or compositions of the present invention (for example, a composition comprising a compound of formula (II)). In some embodiments, administration reduces the patient's risk of recurrent deep vein thrombosis compared to a patient not receiving the compound.

In one aspect, the present invention provides a method for preventing venous thromboembolism, such as deep vein thrombosis or pulmonary embolism, in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (for example, a composition comprising the compound formula (II)). In some embodiments, the patient undergoes surgery. In some embodiments, a compound, a pharmaceutically acceptable salt, or composition thereof is administered to a patient before, during, or after surgery. In some embodiments, the patient undergoes knee or hip replacement surgery. In some embodiments, the patient undergoes orthopedic surgery. In some embodiments, the patient undergoes lung surgery. In some embodiments, the patient is treated for cancer, such as through surgery. In some embodiments, the patient suffers from a chronic disease. In some embodiments, venous thromboembolism is associated with cancer. In some embodiments, the compound, pharmaceutically acceptable salt thereof, or composition of the present invention is a primary medicinal agent in the prevention of deep vein thrombosis or venous thromboembolism. In some embodiments, a compound, a pharmaceutically acceptable salt thereof, or a composition of the present invention is used as an extended therapeutic treatment. In one aspect, the present invention provides a method of reducing a patient's risk of venous thromboembolism, such as deep vein thrombosis or pulmonary embolism, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (for example, a composition comprising compound of formula (II)). In some embodiments, the patient undergoes surgery. In some embodiments, a compound, a pharmaceutically acceptable salt or composition thereof is administered to a patient following surgery. In some embodiments, the patient undergoes knee or hip replacement surgery. In some embodiments, the patient undergoes orthopedic surgery. In some embodiments, the patient undergoes lung surgery. In some embodiments, the patient is treated for cancer, such as through surgery. In some embodiments, the patient suffers from a chronic disease. In some embodiments, the thromboembolic disorder is associated with cancer. In some embodiments, the compound, pharmaceutically acceptable salt or composition thereof of the present invention is a primary medicinal agent in reducing the risk of a thromboembolic disorder. In some embodiments, a compound, a pharmaceutically acceptable salt thereof, or a composition of the present invention is used as an extended therapeutic treatment.

In one aspect, the present invention provides a method of reducing the risk of stroke or systemic embolism in a patient in need thereof, comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention, e.g. a composition comprising a compound of formula (II). In some embodiments, the patient suffers from atrial fibrillation (eg, non-valvular atrial fibrillation). In some embodiments, the patient suffers from impaired renal function (eg, end-stage renal disease).

In one aspect, the present invention provides a method for preventing stroke or systemic embolism in a patient in need thereof, comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt or composition of the present invention, e.g. comprising a compound of formula (II). In some embodiments, the patient suffers from atrial fibrillation (eg, non-valvular atrial fibrillation). In some embodiments, the patient suffers from impaired renal function (eg, end-stage renal failure).

In one aspect, the present invention provides a method of reducing the risk of recurrent pulmonary embolism (eg, symptomatic pulmonary embolism) comprising administering to a patient who has suffered a pulmonary embolism (eg, a patient who has previously been treated for pulmonary embolism) an effective amount of a compound of formula ( II) or a pharmaceutically acceptable salt or composition thereof of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, administration reduces the patient's risk of recurrent pulmonary embolism compared to a patient not administered the compound.

In one aspect, the present invention provides a method for preventing pulmonary embolism in a patient who has suffered a pulmonary embolism (eg, a patient who has previously been treated for pulmonary embolism), comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof according to the present invention (for example, a composition comprising a compound of formula (II)).

In one aspect, the present invention provides a method of reducing the risk of recurrent pulmonary embolism (eg, symptomatic pulmonary embolism) comprising administering to a patient who has suffered deep vein thrombosis (eg, a patient who has previously been treated for deep vein thrombosis) an effective amount of a compound of formula ( II) or a pharmaceutically acceptable salt or composition thereof of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, administration reduces the patient's risk of recurrent pulmonary embolism compared to a patient not administered the compound.

In one aspect, the present invention relates to a method of preventing pulmonary embolism in a patient who has suffered deep vein thrombosis (for example, in a patient who has previously been treated for deep vein thrombosis), comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or compositions of the present invention (for example, a composition comprising a compound of formula (II)).

In one aspect, the present invention relates to a method of treating pulmonary embolism in a patient who has suffered deep vein thrombosis, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention, e.g. a composition comprising a compound of formula (II).

In one aspect, the present invention relates to a method of treating deep vein thrombosis in a patient who has previously been administered an anticoagulant, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (for example, a composition comprising a compound of formula (II) )). In some embodiments, the anticoagulant is administered parenterally over a period of 5-10 days.

In one aspect, the present invention relates to a method of treating pulmonary embolism in a patient who has previously been administered an anticoagulant, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (for example, a composition comprising a compound of formula (II) )). In some embodiments, the anticoagulant is administered parenterally for 5-10 days.

In one aspect, the present invention provides a method of treating a patient who has suffered an ischemic attack (eg, transient ischemic attack), comprising administering a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II) )) to the patient. In some embodiments, the compound is administered to a patient within 24 hours or less, such as 12, 10, 9, 8, 7, 6 hours or less, after the onset of an ischemic attack in the patient.

In one aspect, the present invention provides a method of treating a patient who has suffered an ischemic attack (e.g., transient ischemia), comprising administering a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (e.g., a composition comprising a compound of formula (II) ) to the patient. In some embodiments, the compound is administered to a patient over a period of more than 2 hours to 12 hours, such as more than 2 hours to 10 hours or less, more than 2 hours to 8 hours or less, after the onset of an ischemic attack in the patient.

In one aspect, the present invention provides a method of treating hypertension, e.g., arterial hypertension, in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (e.g., a composition comprising a compound of formula (II)) . In some embodiments, hypertension, such as arterial hypertension, leads to atherosclerosis. In some embodiments, the hypertension is pulmonary arterial hypertension.

In one aspect, the present invention provides a method of reducing the risk of hypertension, such as arterial hypertension, in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (e.g., a composition comprising a compound of formula (II) ). In some embodiments, hypertension, such as arterial hypertension, leads to atherosclerosis. In some embodiments, the hypertension is pulmonary arterial hypertension.

In one aspect, the present invention provides a method for preventing hypertension, e.g., arterial hypertension, in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (e.g., a composition comprising a compound of formula (II)) . In some embodiments, hypertension, such as arterial hypertension, leads to atherosclerosis. In some embodiments, the hypertension is pulmonary arterial hypertension.

In one aspect, the present invention provides a method of reducing inflammation in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, the inflammation is vascular inflammation. In some embodiments, vascular inflammation is accompanied by atherosclerosis. In some embodiments, the patient's vascular inflammation is accompanied by thromboembolic disease. In some embodiments, the vascular inflammation is angiotensin II-induced vascular inflammation.

In one aspect, the present invention provides a method of preventing vascular infiltration of leukocytes in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)).

In one aspect, the present invention provides a method of preventing endothelial dysfunction induced by angiotensin II in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (e.g., a composition comprising a compound of formula (II)) .

In one aspect, the present invention provides a method of preventing excessive thrombin production in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, excess thrombin production occurs on platelets.

In one aspect, the present invention provides a method of treating renal dysfunction associated with hypertension in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (e.g., a composition comprising a compound of formula (II)) .

In one aspect, the present invention provides a method for preventing renal dysfunction associated with hypertension in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (e.g., a composition comprising a compound of formula (II) ).

In one aspect, the present invention provides a method of reducing the risk of renal dysfunction associated with hypertension in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (for example, a composition comprising a compound of formula (II) )).

In one aspect, the present invention provides a method of treating renal fibrosis in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)).

In one aspect, the present invention provides a method for preventing renal fibrosis in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)).

In one aspect, the present invention provides a method of reducing the risk of renal fibrosis in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)).

In one aspect, the present invention provides a method of treating renal failure in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)).

In one aspect, the present invention provides a method for preventing renal failure in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)).

In one aspect, the present invention provides a method of reducing the risk of renal failure in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)). In one aspect, the present invention provides a method of inhibiting factor XIa in a patient, comprising administering to a patient who has suffered ischemia an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (e.g., a composition comprising a compound of formula (II) ). In some embodiments, the ischemia is coronary ischemia. In some embodiments, the patient is a mammal (eg, a human). In some embodiments, the patient undergoes surgery (eg, knee replacement or hip replacement). In some embodiments, the ischemia is coronary ischemia. In some embodiments, the patient is a patient with non-valvular atrial fibrillation. In some embodiments, the patient has one or more of the following risk factors for stroke: a history of stroke (eg, ischemic, unknown, hemorrhagic), transient ischemic attack, or systemic embolism outside the CNS. In some embodiments, the patient has one or more of the following risk factors: age 75 years or more, hypertension, heart failure or left ventricular ejection fraction (eg, less than or equal to 35%), or diabetes mellitus.

In some embodiments, the compound is administered by an oral or parenteral (eg, intravenous) route of administration. In some embodiments, the compound is administered by an oral route.

In some embodiments, the compound is administered prior to an ischemic attack (eg, to a patient at risk for an ischemic attack).

In some embodiments, the compound is administered following an ischemic attack (particularly a transient ischemic attack). In some embodiments, the compound is administered approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days or more after an ischemic attack (particularly a transient ischemic attack). In some embodiments, the compound is administered approximately 1, 2, 3, 4, 5, 6, 7, or 8 weeks or more after an ischemic attack (particularly a transient ischemic attack).

In some embodiments, the compound is administered in combination with an additional therapeutic agent. In some embodiments, the additional therapeutic agent is administered after administration of the compound. In some embodiments, the additional therapeutic agent is administered orally. In some embodiments, the additional therapeutic agent is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 16, 18, 20, or 24 hours or more after administration of the compound. In some embodiments, the additional therapeutic agent is administered at least 1, 2, 3, 4, 5, 6, 7, 14, 21, or 28 days or more after administration of the compound. In some embodiments, the additional therapeutic agent is administered about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days or more after administration of the compound.

In some embodiments, the additional therapeutic agent is administered continuously (e.g., for about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, or about 14 days or more) after administration of the compound.

In some embodiments, the additional therapeutic agent treats a side effect (e.g., active abnormal bleeding or severe hypersensitivity reactions (e.g., anaphylactic reactions), spinal and or epidural hematoma, gastrointestinal distress (e.g., upper abdominal pain, dyspepsia, toothache ), general disorders and injection site conditions (eg, fatigue), infections and infestations (eg, sinusitis, urinary tract infections), musculoskeletal and connective tissue disorders (eg, back pain, osteoarthritis), respiratory disorders , chest and mediastinal organs (for example, pain in the oropharynx), trauma, infection and complications after the procedure (for example, discharge from a wound), disorders of the skeletal muscles and connective tissue (for example, pain in the limb, muscle spasm), disorders of the nervous system systems (eg, syncope), cutaneous and subcutaneous tissue disorders (eg, ruritis, blisters), hematopoietic and lymphatic system disorders (eg, agranulocytosis), gastrointestinal disorders (eg, retroperitoneal hemorrhage), hepatobiliary disorders (eg , jaundice, cholestasis, cytolytic hepatitis), immune system disorders (eg, hypersensitivity, anaphylatic reaction, anaphylactic shock, angioedema), nervous system disorders (eg, cerebral hemorrhage, subdural hematoma, epidural hematoma, hemiparesis), skin and subcutaneous disorders fiber (for example, Stevens-Johnson syndrome)).

In some embodiments, the additional therapeutic agent is a nonsteroidal anti-inflammatory drug (NSAID) (eg, aspirin or naproxen), a platelet aggregation inhibitor (eg, clopidogrel), or an anticoagulant (eg, warfarin or enoxaparin).

In some embodiments, the additional therapeutic agent results in an additive therapeutic effect. In some embodiments, the additional therapeutic agent results in a synergistic therapeutic effect.

In another aspect, the present invention relates to a pharmaceutical composition comprising a compound of the present invention (eg, a compound of formula (II)) and a pharmaceutically acceptable excipient.

In another aspect, the present invention relates to a method of modulating (eg, inhibiting) factor XIa in a patient. The method includes the step of administering an effective amount of a compound of the present invention (for example, a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention (for example, a composition comprising a compound of formula (II)) to a patient in need thereof, and modulating such way (for example, by inhibiting) factor XIa.

In another aspect, the present invention relates to a method of treating a thromboembolic complication in a patient in need thereof. The method involves administering to a patient an effective amount of a compound of the present invention (eg, a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention (eg, a composition comprising a compound of formula (II)). The thromboembolic complication may be arterial cardiovascular thromboembolic complications, arterial thrombosis, venous cardiovascular thromboembolic complications, and thromboembolic complications in the heart chambers, including unstable angina, acute coronary syndrome, primary myocardial infarction, recurrent myocardial infarction, ischemia (eg, coronary ischemia, sudden ischemic death or transient ischemic attack), stroke, atherosclerosis, peripheral arterial occlusive disease, venous thromboembolism, venous thrombosis, deep vein thrombosis, thrombophlebitis, arterial embolism, coronary thrombosis, cerebral artery thrombosis, cerebral embolism, renal embolism, pulmonary embolism and thrombosis resulting from the insertion of (a) artificial valves or other implants, (b) abdominal catheters, (c) stents, (d) cardiopulmonary bypass, (e) hemodialysis, or (f) other procedures in which the blood is exposed to artificial surface, which promotes thrombosis.

In another aspect, the present invention relates to a method for preventing a thromboembolic complication in a patient. The method involves administering to a patient an effective amount of a compound of the present invention (eg, a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention (eg, a composition comprising a compound of formula (II)). The thromboembolic complication may be arterial cardiovascular thromboembolic complications, arterial thrombosis, venous cardiovascular thromboembolic complications, and thromboembolic complications in the cardiac chambers, including unstable angina, acute coronary syndrome, primary myocardial infarction, recurrent myocardial infarction, ischemia (eg, coronary ischemia, sudden ischemic death or transient ischemic attack), stroke, atherosclerosis, peripheral arterial occlusive disease, venous thromboembolism, venous thrombosis, deep vein thrombosis, thrombophlebitis, arterial embolism, coronary artery thrombosis, cerebral artery thrombosis, cerebral embolism, renal embolism, pulmonary embolism and thrombosis, resulting from the insertion of (a) artificial valves or other implants, (b) catheters, (c) stents, (d) cardiopulmonary bypass, (e) hemodialysis or (f) other procedures in which blood is exposed to an artificial surface, which promotes thrombosis.

In another aspect, the present invention provides a method for reducing the risk of a thromboembolic complication in a patient. The method involves administering to a patient an effective amount of a compound of the present invention (eg, a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention (eg, a composition comprising a compound of formula (II)). The thromboembolic complication may be arterial cardiovascular thromboembolic complications, arterial thrombosis, venous cardiovascular thromboembolic complications, and thromboembolic complications in the cardiac chambers, including unstable angina, acute coronary syndrome, primary myocardial infarction, recurrent myocardial infarction, ischemia (eg, coronary ischemia, sudden ischemic death or transient ischemic attack), stroke, atherosclerosis, peripheral arterial occlusive disease, venous thromboembolism, venous thrombosis, deep vein thrombosis, thrombophlebitis, arterial embolism, coronary artery thrombosis, cerebral artery thrombosis, cerebral embolism, renal embolism, pulmonary embolism and thrombosis, resulting from the insertion of (a) artificial valves or other implants, (b) catheters, (c) stents, (d) cardiopulmonary bypass, (e) hemodialysis or (f) other procedures in which blood is exposed to an artificial surface, which promotes thrombosis.

In one aspect, the present invention provides a method of treating end-stage renal disease in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)).

In one aspect, the present invention provides a method for preventing end-stage renal disease in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)).

In one aspect, the present invention provides a method of reducing the risk of end-stage renal disease in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)).

In another aspect, the present invention provides a method of treating a thromboembolic complication in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (e.g., a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention (e.g. , a composition comprising a compound of formula (II)), wherein the patient is exposed to an artificial surface. In some embodiments, the artificial surface is in contact with the patient's blood. In some embodiments, the artificial surface is an extracorporeal surface. In some embodiments, the artificial surface is the surface of an implantable device, such as a mechanical valve. In some embodiments, the artificial surface is the surface of a dialysis catheter. In some embodiments, the artificial surface is a cardiopulmonary bypass circuit. In some embodiments, the artificial surface is the surface of an artificial heart valve. In some embodiments, the artificial surface is the surface of a ventricular support device. In some embodiments, the artificial surface is a small graft surface. In some embodiments, the artificial surface is the surface of a central venous catheter. In some embodiments, the artificial surface is the surface of an extracorporeal membrane oxygenation (ECMO) machine. In some embodiments, the artificial surface causes or is associated with thromboembolic complications. In some embodiments, the thromboembolic complication is a venous thromboembolism. In some embodiments, the thromboembolic complication is deep vein thrombosis. In some embodiments, the thromboembolic complication is a pulmonary embolism.

In another aspect, the present invention provides a method of reducing the risk of a thromboembolic complication in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (e.g., a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention (eg, a composition comprising a compound of formula (II)), where the patient is exposed to an artificial surface. In some embodiments, the artificial surface is in contact with the patient's blood. In some embodiments, the artificial surface is an extracorporeal surface. In some embodiments, the artificial surface is the surface of an implantable device, such as a mechanical valve. In some embodiments, the artificial surface is the surface of a dialysis catheter. In some embodiments, the artificial surface is a cardiopulmonary bypass circuit. In some embodiments, the artificial surface is the surface of an artificial heart valve. In some embodiments, the artificial surface is the surface of a ventricular support device. In some embodiments, the artificial surface is a small graft surface. In some embodiments, the artificial surface is the surface of a central venous catheter. In some embodiments, the artificial surface is the surface of an extracorporeal membrane oxygenation (ECMO) machine. In some embodiments, the artificial surface causes or is associated with thromboembolic complications. In some embodiments, the thromboembolic complication is a venous thromboembolism. In some embodiments, the thromboembolic complication is deep vein thrombosis. In some embodiments, the thromboembolic complication is a pulmonary embolism.

In another aspect, the present invention provides a method for preventing a thromboembolic complication in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (e.g., a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention ( for example, a composition comprising a compound of formula (II)), wherein the patient is exposed to an artificial surface. In some embodiments, the artificial surface is in contact with the patient's blood. In some embodiments, the artificial surface is an extracorporeal surface. In some embodiments, the artificial surface is the surface of an implantable device, such as a mechanical valve. In some embodiments, the artificial surface is the surface of a dialysis catheter. In some embodiments, the artificial surface is a cardiopulmonary bypass circuit. In some embodiments, the artificial surface is the surface of an artificial heart valve. In some embodiments, the artificial surface is the surface of a ventricular support device. In some embodiments, the artificial surface is a small graft surface. In some embodiments, the artificial surface is the surface of a central venous catheter. In some embodiments, the artificial surface is the surface of an extracorporeal membrane oxygenation (ECMO) machine. In some embodiments, the artificial surface causes or is associated with thromboembolic complications. In some embodiments, the thromboembolic complication is a venous thromboembolism. In some embodiments, the thromboembolic complication is deep vein thrombosis. In some embodiments, the thromboembolic complication is a pulmonary embolism.

In another aspect, the present invention provides a method for treating atrial fibrillation in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (e.g., a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention ( for example, a composition comprising a compound of formula (II)). In some embodiments, the patient also requires dialysis, such as kidney dialysis. In some embodiments, a compound of the present invention is administered to a patient during dialysis. In some embodiments, the compound or a pharmaceutically acceptable salt or composition thereof is administered to a patient before or after dialysis. In some embodiments, the patient suffers from end-stage renal disease. In some embodiments, the patient does not require dialysis, such as kidney dialysis. In some embodiments, the patient is at high risk of bleeding. In some embodiments, atrial fibrillation is associated with another thromboembolic complication, such as a blood clot (thrombus).

In another aspect, the present invention provides a method of reducing the risk of atrial fibrillation in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (eg, a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, the patient is at high risk of developing atrial fibrillation. In some embodiments, the patient also requires dialysis, such as kidney dialysis. In some embodiments, a compound of the present invention is administered to a patient during dialysis. In some embodiments, the compound or a pharmaceutically acceptable salt or composition thereof is administered to a patient before or after dialysis. In some embodiments, the patient suffers from end-stage renal disease. In some embodiments, the patient does not require dialysis, such as kidney dialysis. In some embodiments, the patient is at high risk of bleeding. In some embodiments, atrial fibrillation is associated with another thromboembolic complication, such as a blood clot.

In another aspect, the present invention provides a method for preventing atrial fibrillation in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (e.g., a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention ( for example, a composition comprising a compound of formula (II)). In some embodiments, the patient is at high risk of developing atrial fibrillation. In some embodiments, the patient also requires dialysis, such as kidney dialysis. In some embodiments, a compound of the present invention is administered to a patient during dialysis. In some embodiments, the compound or a pharmaceutically acceptable salt or composition thereof is administered to a patient before or after dialysis. In some embodiments, the patient suffers from end-stage renal disease. In some embodiments, the patient does not require dialysis, such as kidney dialysis. In some embodiments, the patient is at high risk of bleeding. In some embodiments, atrial fibrillation is associated with another thromboembolic complication, such as a blood clot.

In another aspect, the present invention provides a method for treating heparin-induced thrombocytopenia in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (e.g., a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof. invention (eg, a composition comprising a compound of formula (II)).

In another aspect, the present invention provides a method of reducing the risk of heparin-induced thrombocytopenia in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (eg, a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof according to the present invention (eg, a composition comprising a compound of formula (II)).

In another aspect, the present invention provides a method for preventing heparin-induced thrombocytopenia in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (e.g., a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof. invention (eg, a composition comprising a compound of formula (II)).

In another aspect, the present invention provides a method for treating heparin-induced thrombocytopenia and thrombosis in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (e.g., a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof according to the present invention (for example, a composition comprising a compound of formula (II)).

In another aspect, the present invention provides a method of reducing the risk of heparin-induced thrombocytopenia and thrombosis in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (eg, a compound of formula (II)) or a pharmaceutically acceptable salt thereof or compositions of the present invention (for example, a composition comprising a compound of formula (II)).

In another aspect, the present invention provides a method for preventing heparin-induced thrombocytopenia and thrombosis in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (e.g., a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof according to the present invention (for example, a composition comprising a compound of formula (II)).

In another aspect, the present invention provides a method for preventing a thromboembolic complication in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (e.g., a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention ( for example, a composition comprising a compound of formula (II)), where the patient has cancer or is undergoing chemotherapy. In some embodiments, the patient simultaneously receives chemotherapy. In some embodiments, the patient has elevated lactate dehydrogenase levels. In some embodiments, the thromboembolic complication is a venous thromboembolism. In some embodiments, the thromboembolic complication is deep vein thrombosis. In some embodiments, the thromboembolic complication is a pulmonary embolism.

In another aspect, the present invention provides a method of treating thrombotic microangiopathy in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (e.g., a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention ( for example, a composition comprising a compound of formula (II)). In some embodiments, the thrombotic microangiopathy is hemolytic uremic syndrome (HUS). In some embodiments, the thrombotic microangiopathy is thrombotic thrombocytopenic purpura (TTP).

In another aspect, the present invention provides a method of reducing the risk of thrombotic microangiopathy in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (e.g., a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, the thrombotic microangiopathy is hemolytic uremic syndrome (HUS). In some embodiments, the thrombotic microangiopathy is thrombotic thrombocytopenic purpura (TTP).

In another aspect, the present invention provides a method for preventing thrombotic microangiopathy in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (e.g., a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention ( for example, a composition comprising a compound of formula (II)). In some embodiments, the thrombotic microangiopathy is hemolytic uremic syndrome (HUS). In some embodiments, the thrombotic microangiopathy is thrombotic thrombocytopenic purpura (TTP).

In another aspect, the present invention provides a method for preventing recurrent ischemia in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention (eg, a compound of formula (II)) or a pharmaceutically acceptable salt or composition thereof of the present invention ( for example, a composition comprising a compound of formula (II)), wherein the patient has an acute coronary syndrome. In some embodiments, the patient suffers from atrial fibrillation. In some embodiments, the patient does not suffer from atrial fibrillation. In another aspect, the present invention provides a method of treating a patient identified as being at risk, eg, high risk, for stroke or thrombosis, which reduces the patient's likelihood of stroke or thrombosis. In some embodiments, the patient is also identified as being at risk for bleeding (eg, heavy bleeding) or sepsis. In some embodiments, the treatment is effective without a tendency to bleed. In some embodiments, treatment is effective in maintaining patency of infusion ports and lines. In addition, the compounds of the present invention (eg, compounds of formula (II)) can be used in the treatment and prevention of other diseases in which thrombin generation plays a physiological role. For example, thrombin contributes to morbidity and mortality from chronic and degenerative diseases such as cancer, arthritis, atherosclerosis, vascular dementia and Alzheimer's disease due to its ability to regulate the function of various cell types through specific cleavage and activation of the cell surface thrombin receptor, mitogenic effects, various cellular functions such as cellular proliferation, for example abnormal proliferation of vascular cells leading to restenosis or angiogenesis, PDGF release and DNA synthesis. Inhibition of factor XIa effectively blocks the generation of thrombin and therefore neutralizes any physiological effects of thrombin on various cell types. The typical indications discussed above include some, but not all, potential clinical situations amenable to treatment with a factor XIa inhibitor.

In another aspect, the present invention relates to a method of treating a patient who has edema (for example, angioedema, in particular hereditary angioedema), comprising administering a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention (for example, a composition comprising a compound of formula (II)) to the patient.

In another aspect, the present invention relates to a method of preventing edema (for example, allergic angioedema, in particular hereditary angioedema) in a patient, comprising administering a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (for example, a composition comprising the compound formula (II)) to the patient.

In another aspect, the present invention relates to a method of reducing the risk of edema (for example, angioedema, in particular hereditary angioedema) in a patient, comprising administering a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (for example, a composition comprising the compound formula (II)) to the patient.

In another aspect, the present invention provides a method of inhibiting kallikrein in a patient, comprising administering to a patient with edema (e.g., angioedema, particularly hereditary angioedema) an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention (e.g. composition comprising a compound of formula (II)).

In another aspect, the present invention provides a method of treating a thromboembolic consequence or complication in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, the thromboembolic consequence or complication is associated with intervention in the peripheral blood vessels (eg, extremities), hemodialysis, catheter ablation, cerebrovascular intervention, organ transplantation (eg, liver), surgery (eg, orthopedic surgery, pulmonary surgery, surgery abdominal or cardiac surgery (eg, open heart surgery), transcatheter aortic valve implantation, large-bore surgery used to treat an aneurysm, percutaneous coronary intervention, or hemophilia therapy. In some embodiments, the surgery is orthopedic surgery, pulmonary surgery, abdominal surgery, or cardiac surgery.

In another aspect, the present invention provides a method for preventing a thromboembolic consequence or complication in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, the thromboembolic consequence or complication is associated with intervention in the peripheral blood vessels (eg, extremities), hemodialysis, catheter ablation, cerebrovascular intervention, organ transplantation (eg, liver), surgery (eg, orthopedic surgery, pulmonary surgery, surgery abdominal or cardiac surgery (eg, open heart surgery), transcatheter aortic valve implantation, large-bore surgery used to treat an aneurysm, percutaneous coronary intervention, or hemophilia therapy. In some embodiments, the surgery is orthopedic surgery, pulmonary surgery, abdominal surgery, or cardiac surgery.

In another aspect, the present invention provides a method of reducing the risk of a thromboembolic consequence or complication in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, the thromboembolic consequence or complication is associated with intervention in the peripheral blood vessels (eg, extremities), hemodialysis, catheter ablation, cerebrovascular intervention, organ transplantation (eg, liver), surgery (eg, orthopedic surgery, pulmonary surgery, surgery abdominal or cardiac surgery (eg, open heart surgery), transcatheter aortic valve implantation, large-bore surgery used to treat an aneurysm, percutaneous coronary intervention, or hemophilia therapy. In some embodiments, the surgery is orthopedic surgery, pulmonary surgery, abdominal surgery, or cardiac surgery.

In another aspect, the invention provides a method of treating restenosis following arterial injury in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, arterial injury occurs following cranial artery stenting.

In another aspect, the present invention provides a method for preventing restenosis following arterial injury in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, arterial injury occurs following cranial artery stenting.

In another aspect, the present invention provides a method of reducing the risk of restenosis after arterial injury in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, arterial injury occurs following cranial artery stenting.

In another aspect, the present invention provides a method of treating hepatic vascular thrombosis in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)).

In another aspect, the present invention provides a method for preventing hepatic vascular thrombosis in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)).

In another aspect, the present invention provides a method of reducing the risk of hepatic vascular thrombosis in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)).

In another aspect, the present invention provides a method of treating non-ST segment elevation myocardial infarction or ST segment elevation myocardial infarction, comprising administering to a patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention (e.g., a composition comprising a compound of formula (II)).

In another aspect, the present invention provides a method for preventing non-ST segment elevation myocardial infarction or ST segment elevation myocardial infarction in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention (e.g. composition comprising a compound of formula (II)).

In another aspect, the present invention provides a method of reducing the risk of non-ST segment elevation myocardial infarction or ST segment elevation myocardial infarction in a patient, comprising administering to the patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention (eg , a composition comprising a compound of formula (II)).

In another aspect, the present invention relates to a method of maintaining blood vessel patency, comprising administering to a patient an effective amount of a compound of formula (II) or a pharmaceutically acceptable salt thereof or a composition of the present invention (eg, a composition comprising a compound of formula (II)). In some embodiments, the patient is suffering from acute renal failure. In some embodiments, the patient additionally receives continuous renal replacement therapy.

In some of the above embodiments, the compound of the present invention or composition thereof is administered orally or parenterally. In some embodiments, the compound or composition thereof is administered orally. In some embodiments, the compound or composition thereof is administered after the patient has stopped using a direct oral anticoagulant. In some embodiments, the patient has been using the oral coagulant for approximately 2.5 years. In some embodiments, the patient is a mammal, such as a human.

DETAILED DESCRIPTION

Definitions

The term "alkyl", by itself or as part of another substituent, means, unless otherwise stated, a straight or branched chain or cyclic hydrocarbon radical or combination thereof, which may be fully saturated, mono- or polyunsaturated and may include di- and multivalent radicals containing a specified number of carbon atoms (ie, C ₁ -C ₁₀ means from one to ten carbon atoms). Examples of saturated hydrocarbon radicals include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, homologues and isomers , for example, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc. An unsaturated alkyl group is a group containing one or more double bonds or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3 -propynyl, 3-butynyl and their higher homologues and isomers. The term "alkyl", unless otherwise specified, also includes alkyl derivatives defined in more detail below, such as "heteroalkyl". Alkyl groups that are limited to hydrocarbon groups are called "homoalkyl". Unless otherwise indicated, each example of an alkyl group is optionally substituted, i.e. unsubstituted (“unsubstituted alkyl”) or substituted (“substituted alkyl”) with one or more substituents; for example 1-5 substituents, 1-3 substituents or 1 substituent. In some embodiments, the alkyl group is an unsubstituted C _1-10 alkyl (eg, -CH ₃ ). In some embodiments, the alkyl group is a substituted C _1-10 alkyl. Common abbreviations include Me (-CH ₃ ), Et (-CH ₂ CH ₃ ), iPr (-CH(CH ₃ ) ₂ ), nPr (-CH ₂ CH ₂ CH ₃ ), n-Bu (-CH ₂ CH ₂ CH ₂ CH ₃ ) or i-Bu (-CH ₂ CH(CH ₃ ) ₂ ).

The term “alkylene,” by itself or as part of another substituent, means a divalent radical derived from an alkane, such as, but not limited to, -CH ₂ CH ₂ CH ₂ CH ₂ -, and further includes the groups described below as “heteroalkylene”. Typically, an alkyl (or alkylene) group will contain from 1 to 24 carbon atoms, with groups containing 10 or fewer carbon atoms being preferred in the present invention. "Lower alkyl" or "lower alkylene" is a shorter chain alkyl or alkylene group, typically containing eight or fewer carbon atoms.

The term "alkenyl" refers to a straight or branched hydrocarbon chain containing 2-12 carbon atoms (unless otherwise noted) and containing one or more double bonds. Examples of alkenyl groups include, but are not limited to, allylic, propenyl, 2-butenyl, 3-hexenyl and 3-octenyl groups. One of the carbon atoms of the double bond may optionally be the point of attachment of the alkenyl substituent.

The term "alkenylene" refers to divalent alkenyl, for example -CH=CH-, -CH ₂ -CH=CH- and -CH=CH-CH ₂ -.

The term "alkynyl" refers to a straight or branched hydrocarbon mixture containing 2-12 carbon atoms (unless otherwise specified) and characterized by the presence of one or more triple bonds. Examples of alkynyl groups include, but are not limited to, ethynyl, propargyl and 3-hexynyl. One of the carbon atoms of the triple bond may optionally be the point of attachment of the alkynyl substituent.

The term "alkynylene" refers to divalent alkynyl, for example -CH≡CH-, -CH ₂ -CH≡CH- and -CH≡CH-CH ₂ -.

The terms "alkoxy", "alkylamino" and "alkylthio" (or "thioalkoxy") are used in their usual meaning and refer to alkyl groups attached to the rest of the molecule through an oxygen atom, an amino group or a sulfur atom, respectively.

The terms "cyano" and "nitrile" refer to the -CN radical.

The terms "cycloalkyl", "heterocycloalkyl" or "heterocyclyl", alone or in combination with other terms, mean, unless otherwise indicated, the cyclic variants of "alkyl" and "heteroalkyl", respectively. Additionally, in a heterocycloalkyl or heterocyclyl, the heteroatom may occupy a position in which the heterocycle is attached to the rest of the molecule. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, cyclooctanyl, and the like. Examples of heterocycloalkyl and heterocyclyl include, but are not limited to, 1-1,2,5,6-tetrahydropyridyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran- 3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, 2-tetrahydropyranyl, 3-tetrahydropyranyl, 4-tetrahydropyranyl, 1-methyl-1,5-dihydro-2-imidazol- 4-onyl, etc.

The term “heteroalkyl” as used herein refers to an alkyl group as defined herein further including 1 or more (e.g., 1, 2, 3, or 4) heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus) in the main chain, with either several heteroatoms inserted between adjacent carbon atoms in the main carbon chain and/or one or more heteroatoms inserted between a carbon atom and the main molecule, i.e. between the connection point. In some embodiments, a heteroalkyl group refers to a saturated group containing 1 to 10 carbon atoms and 1, 2, 3, or 4 heteroatoms (“hetero-C _1-10 alkyl”). In some embodiments, a heteroalkyl group is a saturated group containing 1 to 9 carbon atoms and 1, 2, 3, or 4 heteroatoms (“hetero-C _1-9 alkyl”). In some embodiments, a heteroalkyl group is a saturated group containing 1 to 8 carbon atoms and 1, 2, 3, or 4 heteroatoms (“hetero-C _1-8 alkyl”). In some embodiments, a heteroalkyl group is a saturated group containing 1 to 7 carbon atoms and 1, 2, 3, or 4 heteroatoms (“hetero-C _1-7 alkyl”). In some embodiments, a heteroalkyl group is a group containing 1 to 6 carbon atoms and 1, 2, or 3 heteroatoms (“hetero-C _1-6 alkyl”). In some embodiments, a heteroalkyl group is a saturated group containing 1 to 5 carbon atoms and 1 or 2 heteroatoms (“hetero-C _1-5 alkyl”). In some embodiments, a heteroalkyl group is a saturated group containing 1 to 4 carbon atoms and 1 or 2 heteroatoms (“hetero-C _1-4 alkyl”). In some embodiments, a heteroalkyl group is a saturated group containing 1 to 3 carbon atoms and 1 heteroatom (“hetero-C _1-3 alkyl”). In some embodiments, a heteroalkyl group is a saturated group containing 1 or 2 carbon atoms and 1 heteroatom (“hetero-C _1-2 alkyl”). In some embodiments, a heteroalkyl group is a saturated group containing 1 carbon atom and 1 heteroatom (“hetero-C ₁ alkyl”). In some embodiments, a heteroalkyl group is a saturated group containing 2 to 6 carbon atoms and 1 or 2 heteroatoms (“hetero-C _2-6 alkyl”). Unless otherwise indicated, each example of a heteroalkyl group is independently unsubstituted (“unsubstituted heteroalkyl”) or substituted (“substituted heteroalkyl”) with one or more substituents. In some embodiments, the heteroalkyl group is unsubstituted hetero-C _1-10 alkyl. In some embodiments, the heteroalkyl group is a substituted hetero-C _1-10 alkyl.

The term "heterocyclyl", when used in combination with other terms (eg, heterocyclylalkyl), includes heterocyclyl rings as defined above. Thus, the term “heterocyclylalkyl” includes radicals in which a heterocyclyl group is attached to an alkyl group, including alkyl groups in which a carbon atom (eg, a methylene group) is replaced by an oxygen atom.

The terms "halogen", by itself or as part of another substituent, means, unless otherwise indicated, a fluorine, chlorine, bromine or iodine atom.

The term "haloalkyl" as used herein refers to an alkyl group as defined herein further including 1 or more (e.g., 1, 2, 3, or 4) halogen atoms (e.g., fluorine, chlorine, bromine, or iodine ), where the alkyl group is substituted with one or more halogen atoms. In some embodiments, a haloalkyl group refers to a saturated group containing 1 to 10 carbon atoms and 1, 2, 3, or 4 halogen atoms (“halo-C _1-10 alkyl”). In addition, the term "haloalkyl" includes monohaloalkyl and polyhaloalkyl. For example, the term "haloalkyl" includes, but is not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

The terms "haloalkoxy" as used herein refers to an alkoxy group as defined herein further including 1 or more (e.g., 1, 2, 3, or 4) halogen atoms (e.g., fluorine, chlorine, bromine, or iodine), where the alkoxy group is substituted with one or more halogen atoms.

The term "hydroxyl" refers to the -OH radical.

The term "aryl" means, unless otherwise stated, a polyunsaturated aromatic hydrocarbon substituent that may be a single ring or include multiple rings (preferably 1 to 3 rings) that are fused or covalently linked. The term "heteroaryl" refers to aryl groups (or rings) that contain from one to four heteroatoms selected from N, O and S atoms, where the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized(s). The heteroaryl group can be attached to the rest of the molecule through a heteroatom. Non-limiting examples of aryl and heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, pyrazinyl, 2-oxazolyl , 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4 -triazolyl, 3-isothiazolyl, 4-isothiazolyl, 5-isothiazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 1,3,4-thiadiazolyl, 3-pyrazolyl, 4-pyrazolyl, 5-pyrazolyl, 2-thienyl , 3-thienyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl , 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, 6-quinolyl and tetrazolyl. Each of the above aryl or heteroaryl ring systems may be substituted, and the substituents are selected from the group of suitable substituents described below. Examples of substituted pyrazolyl include, but are not limited to, 1-methyl-3-pyrazolyl, 1-methyl-4-pyrazolyl, 1-methyl-5-pyrazolyl; examples of substituted imidazolyl include, but are not limited to, 1-methyl-2-imidazolyl, 1-methyl-4-imidazolyl, and 1-methyl-5-imidazolyl. For brevity, the term “aryl,” when used in combination with other terms (eg, aryloxy, arylthio, arylalkyl, aralkyl, heteroaralkyl), includes aryl and heteroaryl rings as described above. Thus, the terms "arylalkyl", "aralkyl" and "heteroaralkyl" include radicals in which an aryl or heteroaryl group is attached to an alkyl group (for example, benzyl, phenethyl, pyridylmethyl, etc.), including alkyl groups in which an atom carbon (eg, a methylene group) is replaced, for example, by an oxygen atom (eg, phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, etc.).

The term "nitro" refers to the -NO ₂ radical.

The term "protecting group" as used herein refers to a portion of a compound that is substantially stable under particular reaction conditions, but which is removed from the compound under other reaction conditions. The protecting group may also be selected such that it participates in direct oxidation of the aromatic ring component of the compounds of the present invention. Examples of protecting groups useful in accordance with the present invention are presented in Greene et al., Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.

Alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups as defined herein are optionally substituted (e.g., "substituted" or "unsubstituted" alkyl, "substituted" or "unsubstituted" alkenyl, "substituted" or " unsubstituted" alkynyl, "substituted" or "unsubstituted" cycloalkyl, "substituted" or "unsubstituted" heterocyclyl, "substituted" or "unsubstituted" aryl or "substituted" or "unsubstituted" heteroaryl group). Generally, the term "substituted", whether preceded by the term "optionally" or not, means that at least one hydrogen atom present on a group (for example, a carbon or nitrogen atom) is replaced by an acceptable substituent, such as a substituent that produces a stable compound, eg a compound that does not undergo spontaneous transformation such as rearrangement, cyclization, elimination, or other reaction. Unless otherwise specified, in a "substituted" group the substituent is at one or more substitutable positions, and when there is more than one substitutable position in any given group, the substituents at each position are the same or different. The term “substituted” is intended to include substitution with all allowable substituents of the organic compounds of the present invention, which results in the formation of a stable compound. For purposes of the present invention, heteroatoms, such as a nitrogen atom, may have hydrogen substituents and/or any suitable substituent, as described herein, that satisfies the valencies of the heteroatoms and results in the formation of a stable ftagment.

Examples of carbon atom substituents include, but are not limited to, halogen, -CN, -NO ₂ , -N ₃ , -SO ₂ H, -SO ₃ H, -OH, -OR ^aa , -ON(R ^bb ) ₂ , -N (R ^bb ) ₂ , -N(R ^bb ) ₃ ⁺ X ^- , -N(OR ^cc )R ^bb , -SH, -SR ^aa , -SSR ^cc , -C(=O)R ^aa , -CO ₂ H , -CHO, -C(OR ^cc ) ₂ , -CO ₂ R ^aa , -OC(=O)R ^aa , -OCO ₂ R ^aa , -C(=O)N(R ^bb ) ₂ , -OC(= O)N(R ^bb ) ₂ , -NR ^bb C(=O)R ^aa , -NR ^bb CO ₂ R ^aa , -NR ^bb C(=O)N(R ^bb ) ₂ , -C(=NR ^bb ) R ^aa , -C(=NR ^bb )OR ^aa , -OC(=NR ^bb )R ^aa , -OC(=NR ^bb )OR ^aa , -C(=NR ^bb )N(R ^bb ) ₂ , -OC( =NR ^bb )N(R ^bb ) ₂ , -NR ^bb C(=NR ^bb )N(R ^bb ) ₂ , -C(=O)NR ^bb SO ₂ R ^aa , -NR ^bb SO ₂ R ^aa , -SO ₂ N(R ^bb ) ₂ , -SO ₂ R ^aa , -SO ₂ OR ^aa , -OSO ₂ R ^aa , -S(O)R ^aa -S(=O)R ^aa , -OS(=O)R ^aa , -Si(R ^aa ) ₃ , -Osi(R ^aa ) ₃ -C(=S)N(R ^bb ) ₂ , -C(=O)SR ^aa , -C(=S)SR ^aa , -SC( =S)SR ^aa , -SC(=O)SR ^aa , -OC(=O)SR ^aa , -SC(=O)OR ^aa , -SC(=O)R ^aa , -P(=O) ₂ R ^aa , -OP(=O) ₂ R ^aa , -P(=O)(R ^aa ) ₂ , -OP(=O)(R ^aa ) ₂ , -OP(=O)(OR ^cc ) ₂ , -P (=O) ₂ N(R ^bb ) ₂ , -OP(=O) ₂ N(R ^bb ) ₂ , -P(=O)(NR ^bb ) ₂ , -OP(=O)(NR ^bb ) ₂ , -NR ^bb P(=O)(OR ^cc ) ₂ , -NR ^bb P(=O)(NR ^bb ) ₂ , -P(R ^cc ) ₂ , -P(R ^cc ) ₃ , -OP(R ^cc ) ₂ , -OP(R ^cc ) ₃ , -B(R ^aa ) ₂ , -B(OR ^cc ) ₂ , -BR ^aa (OR ^cc ), C _1-10 alkyl, C _1-10 perhaloalkyl, C _2-10 alkenyl, _C2-10 alkynyl, _C3-10 cycloalkyl, 3-14 membered heterocyclyl, _C6-14 aryl and 5-14 membered heteroaryl, wherein alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently substituted 0, 1, 2, 3, 4 or 5 groups R ^dd ;

each example of R ^aa is independently selected from C _1-10 alkyl, C _1-10 perhaloalkyl, C _2-10 alkenyl, C _2-10 alkynyl, C _3-10 cycloalkyl, 3-14 membered heterocyclyl, C _6-14 aryl and 5 -14 membered heteroaryl, or two R ^aa groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, where each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 groups R ^dd ;

each example R ^bb is independently selected from hydrogen, -OH, -OR ^aa , -N(R ^cc ) ₂ , -CN, -C(=O)R ^aa , -C(=O)N(R ^cc ) ₂ , - CO ₂ R ^aa , -SO ₂ R ^aa , -C(=NR ^cc )OR ^aa , -C(=NR ^cc )N(R ^cc ) ₂ , -SO ₂ N(R ^cc ) ₂ , -SO ₂ R ^cc , -SO ₂ OR ^cc , -SOR ^aa , -C(=S)N(R ^cc ) ₂ , -C(=O)SR ^cc , -C(=S)SR ^cc , -P(=O) ₂ R ^aa , -P(=O)(R ^aa ) ₂ , -P(=O) ₂ N(R ^cc ) ₂ , -P(=O)(NR ^cc ) ₂ , C _1-10 alkyl, C _1-10 perhaloalkyl, C _2-10 alkenyl, C _2-10 alkynyl, C _3-10 cycloalkyl, 3-14 membered heterocyclyl, C _6-14 aryl and 5-14 membered heteroaryl, or two R ^bb groups joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl ring is independently substituted with 0, 1, 2, 3, 4 or 5 R ^dd groups;

each example R ^cc is independently selected from hydrogen, C _1-10 alkyl, C _1-10 perhaloalkyl, C _2-10 alkenyl, C _2-10 alkynyl, C _3-10 cycloalkyl, 3-14 membered heterocyclyl, C _6-14 aryl and a 5-14 membered heteroaryl, or two R ^cc groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 groups R ^dd ;

each example R ^dd is independently selected from halogen, -CN, -NO ₂ , -N ₃ , -SO ₂ H, -SO ₃ H, -OH, -OR ^ee , -ON(R ^ff ) ₂ , -N(R ^ff ) ₂ , -N(R ^ff ) ₃ ⁺ X ^- , -N(OR ^ee )R ^ff , -SH, -SR ^ee , -SSR ^ee , -C(=O)R ^ee , -CO ₂ H, -CO ₂ R ^ee , -OC(=O)R ^ee , -OCO ₂ R ^ee , -C(=O)N(R ^ff ) ₂ , -OC(=O)N(R ^ff ) ₂ , -NR ^ff C( =O)R ^ee , -NR ^ff CO ₂ R ^ee , -NR ^ff C(=O)N(R ^ff ) ₂ , -C(=NR ^ff )OR ^ee , -OC(=NR ^ff )R ^ee , - OC(=NR ^ff )OR ^ee , -C(=NR ^ff )N(R ^ff ) ₂ , -OC(=NR ^ff )N(R ^ff ) ₂ , -NR ^ff C(=NR ^ff )N(R ^ff ) ₂ ,-NR ^ff SO ₂ R ^ee , -SO ₂ N(R ^ff ) ₂ , -SO ₂ R ^ee , -SO ₂ OR ^ee , -OSO ₂ R ^ee , -S(=O)R ^ee , -Si (R ^ee ) ₃ , -Osi(R ^ee ) ₃ , -C(=S)N(R ^ff ) ₂ , -C(=O)SR ^ee , -C(=S)SR ^ee , -SC(=S )SR ^ee , -P(=O) ₂ R ^ee , -P(=O)(R ^ee ) ₂ , -OP(=O)(R ^ee ) ₂ , -OP(=O)(OR ^ee ) ₂ , C _1-6 alkyl, C _1-6 perhaloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl, C _6-10 aryl, 5-10 membered heteroaryl, where each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 R ^gg groups, or two terminal R ^dd substituents can combine to form =O or =S;

each example R ^ee is independently selected from C _1-6 alkyl, C _1-6 perhaloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-10 cycloalkyl, C _6-10 aryl, 3-10 membered heterocyclyl and 3 -10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R ^gg groups;

each example R ^ff is independently selected from hydrogen, C _1-6 alkyl, C _1-6 perhaloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl, C _6-10 aryl and a 5-10 membered heteroaryl, or two R ^ff groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 groups R ^gg ; And

each example of R ^gg is independently halogen, -CN, -NO ₂ , -N ₃ , -SO ₂ H, -SO ₃ H, -OH, -OC _1-6 alkyl, -ON(C _1-6 alkyl) ₂ , -N(C _1-6 alkyl) ₂ , -N(C _1-6 alkyl) ₃ ⁺ X ^- , -NH(C _1-6 alkyl) ₂ ⁺ X ^- , -NH ₂ (C _1-6 alkyl) ⁺ X ^- , -NH ₃ ⁺ X ^- , -N(OC _1-6 alkyl)(C _1-6 alkyl), -N(OH)(C _1-6 alkyl), -NH(OH), -SH, -SC _1-6 alkyl, -SS(C _1-6 alkyl), -C(=O)(C _1-6 alkyl), -CO ₂ H, -CO ₂ (C _1-6 alkyl), -OC( =O)(C _1-6 alkyl), -OCO ₂ (C _1-6 alkyl), -C(=O)NH ₂ , -C(=O)N(C _1-6 alkyl) ₂ , -OC( =O)NH(C _1-6 alkyl), -NHC(=O)(C _1-6 alkyl), -N(C _1-6 alkyl)C(=O)(C _1-6 alkyl), -NHCO ₂ (C _1-6 alkyl), -NHC(=O)N(C _1-6 alkyl) ₂ , -NHC(=O)NH(C _1-6 alkyl), -NHC(=O)NH ₂ , - C(=NH)O(C _1-6 alkyl), -OC(=NH)(C _1-6 alkyl), -OC(=NH)OC _1-6 alkyl, -C(=NH)N(C _{1 -6} alkyl) ₂ , -C(=NH)NH(C _1-6 alkyl), -C(=NH)NH ₂ , -OC(=NH)N(C _1-6 alkyl) ₂ , -OC(NH )NH(C _1-6 alkyl), -OC(NH)NH ₂ , -NHC(NH)N(C _1-6 alkyl) ₂ , -NHC(=NH)NH ₂ , -NHSO ₂ (C _1-6 alkyl), -SO ₂ N(C _1-6 alkyl) ₂ , -SO ₂ NH(C _1-6 alkyl), -SO ₂ NH ₂ , -SO ₂ C _1-6 alkyl, -SO ₂ OC _1-6 alkyl, -OSO ₂ C _1-6 alkyl, -SOC _1-6 alkyl, -Si(C _1-6 alkyl) ₃ , -OSi(C _1-6 alkyl) ₃ -C(=S)N(C _{1- 6} alkyl) ₂ , C(=S)NH(C _1-6 alkyl), C(=S)NH ₂ , -C(=O)S(C _1-6 alkyl), -C(=S)SC _{1 -6} alkyl, -SC(=S)SC _1-6 alkyl, -P(=O) ₂ (C _1-6 alkyl), -P(=O)(C _1-6 alkyl) ₂ , -OP(= O)(C _1-6 alkyl) ₂ , -OP(=O)(OC _1-6 alkyl) ₂ , C _1-6 alkyl, C _1-6 perhaloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-10 cycloalkyl, C _6-10 aryl, 3-10 membered heterocyclyl, 5-10 membered heteroaryl; or two terminal substituents R ^gg can combine to form =O or =S; where X ^- represents a counterion.

These and other typical examples of substituents are described in more detail in the Detailed Description of the Invention, Examples and Claims.

Connections

The compounds of the present invention are compounds that inhibit factor XIa or kallikrein.

In one aspect, the compound of the present invention is a compound of formula (II):

or a pharmaceutically acceptable salt thereof, wherein R ¹ is hydrogen or -NR ⁸ R ⁹ ; R ^a is C _1-6 alkyl, C _1-6 haloalkyl, halogen, cyano or -OR ⁶ ; R ^b represents hydrogen or C _1-6 alkyl; R ² represents an optionally substituted 5-membered heteroaryl or an optionally substituted 5-membered heterocyclyl; R ³ represents hydrogen, C _1-6 alkyl, or C _1-6 haloalkyl; R ⁴ is C _1-6 alkyl, C _1-6 haloalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted with one, two or three independent substituents selected from halogen, C _{1- 6} alkyl, C _1-6 haloalkyl, cyano or -OR ⁶ ; R ⁵ is hydrogen, C _1-6 alkyl, C _1-6 haloalkyl, cycloalkyl or aryl, wherein cycloalkyl or aryl is optionally substituted with one, two or three independent substituents selected from halogen, C _1-6 alkyl, C _{1- 6} haloalkyl, cyano or -OR ⁶ , or R ⁴ and R ⁵ together with the carbon atom to which they are attached form a ring; R ⁶ represents hydrogen, C _1-6 alkyl, or C _1-6 haloalkyl; each R ⁸ and R ⁹ independently represents hydrogen, C _1-6 alkyl, -C(O)R ¹⁰ , or -C(O)OR ¹⁰ ; R ¹⁰ represents C _1-6 alkyl or C _1-6 haloalkyl; R ^x is -O or absent, where when R ^x is -O, the nitrogen atom of the pyridyl ring is positively charged and R ^x is negatively charged, thereby forming a pyridine N-oxide; m is 0, 1, 2 or 3; and n is 0 or 1, where if n is 0, then R ⁵ is hydrogen and R ⁴ is absent.

In one aspect, the present invention provides a compound of formula (II):

or a pharmaceutically acceptable salt thereof, wherein R ¹ is hydrogen or -NR ⁸ R ⁹ ; R ^a is C _1-6 alkyl, C _1-6 haloalkyl, halogen, cyano or -OR ⁶ ; R ^b represents hydrogen or C _1-6 alkyl; R ² represents an optionally substituted 5-membered heteroaryl; R ³ represents hydrogen, C _1-6 alkyl or C _1-6 haloalkyl; R ⁴ is C _1-6 alkyl, C _1-6 haloalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted with one, two or three independent substituents selected from halogen, C _{1- 6} alkyl, C _1-6 haloalkyl, cyano or -OR ⁶ ; R ⁵ is hydrogen, C _1-6 alkyl, C _1-6 haloalkyl, cycloalkyl or aryl, wherein cycloalkyl or aryl is optionally substituted with one, two or three independent substituents selected from halogen, C _1-6 alkyl, C _{1- 6} haloalkyl, cyano or -OR ⁶ , or R ⁴ and R ⁵ together with the carbon atom to which they are attached form a ring; R ⁶ represents hydrogen, C _1-6 alkyl or C _1-6 haloalkyl; R ⁸ and R ⁹ are each independently hydrogen, C _1-6 alkyl, -C(O)R ¹⁰ or -C(O)OR ¹⁰ ; R ¹⁰ represents C _1-6 alkyl or C _1-6 haloalkyl; m is 0, 1, 2 or 3; and n is 0 or 1, where if n is 0, then R ⁵ is hydrogen and R ⁴ is absent.

In some embodiments, R ^x is absent.

In some embodiments, the compound is a compound of formula (I):

In some embodiments, R ⁶ is C _1-6 alkyl or C _1-6 haloalkyl. In some embodiments, R ⁶ is -CH ₃ or -CF ₃ .

In some embodiments, R ¹ is hydrogen, -NH ₂ or -NHCH ₃ . In some embodiments, R ¹ is hydrogen. In some embodiments, R ¹ is -NH ₂ . In some embodiments, R ¹ is -NHCH ₃ .

In some embodiments, the 5-membered heterocyclyl is a nitrogen-containing 5-membered heterocyclyl. In some embodiments, the nitrogen-containing 5-membered heterocyclyl is saturated or unsaturated.

In some embodiments, R ² is a 5-membered heterocyclyl, optionally substituted with one, two, three or four independent substituents selected from C _1-6 alkyl, C _2-6 alkenyl, C ₂ -C ₆ alkynyl, C _1-6 alkoxyl, C _1-6 haloalkyl, C _1-6 haloalkoxy, halogen, cyano, C _3-8 carbocyclyl, C _3-8 heterocyclyl, aryl or heteroaryl. In some embodiments, the nitrogen-containing 5-membered heterocyclyl is optionally substituted with an oxo group (=O). In some embodiments, R ² is a 5-membered heterocyclyl, optionally substituted with one or two independent substituents selected from C _1-6 alkyl.

In some embodiments, R ² is a 5-membered heteroaryl, optionally substituted with one, two, three or four independent substituents selected from C _1-6 alkyl, C _2-6 alkenyl, C ₂ -C ₆ alkynyl, C _1-6 alkoxyl, C _1-6 haloalkyl, C _1-6 haloalkoxy, halogen, cyano, C _3-8 carbocyclyl, C _3-8 heterocyclyl, aryl or heteroaryl.

In some embodiments, R ² is a 5-membered heteroaryl, optionally substituted with one or two independent substituents selected from C _1-6 alkyl. In some embodiments, R ² is pyrazolyl, imidazolyl, thienyl, isothiazolyl, or thiazolyl, wherein imidazolyl, thienyl, isothiazolyl, or thiazolyl is optionally substituted with one or two independent substituents selected from C _1-6 alkyl. In some embodiments, R ² is pyrazolyl, imidazolyl, thienyl, isothiazolyl, thiazolyl, oxadiazolyl, or thiadiazolyl, wherein imidazolyl, thienyl, isothiazolyl, thiazolyl, oxadiazolyl, or thiadiazolyl is optionally substituted with one or two independent substituents selected from C _1-6 alkyl . In some embodiments, R ² is pyrazolyl, imidazolyl, thienyl, isothiazolyl, or thiazolyl, wherein imidazolyl, thienyl, isothiazolyl, or thiazolyl is optionally substituted with one or two substituents selected from -CH ₃ . In some embodiments, R ² is pyrazolyl, imidazolyl, thienyl, isothiazolyl, thiazolyl, oxadiazolyl, or thiadiazolyl, wherein imidazolyl, thienyl, isothiazolyl, thiazolyl, oxadiazolyl, or thiadiazolyl is optionally substituted with one or two substituents selected from -CH ₃ .

In some embodiments, R ² is

In some embodiments, R ² is

.

In some embodiments, R ² is

In some embodiments, R ² is

In some embodiments, R ² is

In some embodiments, R ³ is C _1-6 alkyl. In some embodiments, R ³ is -CH ₃ .

In some embodiments, R ⁴ is C _1-6 alkyl, cycloalkyl, aryl or heteroaryl, wherein the cycloalkyl, aryl or heteroaryl is optionally substituted with one, two or three independent substituents selected from halogen or C _1-6 alkyl. In some embodiments, R ⁴ is cyclohexyl, cyclopropyl, phenyl, pyridyl, pyrazolyl, thienyl, or C _1-6 alkyl, wherein cyclohexyl, cyclopropyl, phenyl, pyridyl, pyrazolyl, or thienyl is optionally substituted with one, two, or three independent substituents selected from halogen or C _1-6 alkyl. In some embodiments, R ⁴ is cyclohexyl, phenyl, pyridyl, pyrazolyl, or thienyl, wherein the cyclohexyl, phenyl, pyridyl, pyrazolyl, or thienyl is optionally substituted with one, two, or three independent substituents selected from halogen or C _1-6 alkyl. In some embodiments, R ⁴ is phenyl, optionally substituted with one, two or three substituents selected from -F, -Cl or C _1-6 alkyl.

In some embodiments, R ⁴ is C _1-6 alkyl, C _1-6 haloalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted with one, two or three independent substituents selected from halogen , C _1-6 alkyl, C _1-6 haloalkyl, cyano or -OR ⁶ , and R ⁵ is hydrogen, C _1-6 alkyl, C _1-6 haloalkyl, cycloalkyl or aryl, wherein cycloalkyl or aryl is optionally substituted with one , two or three independent substituents selected from halogen, C _1-6 alkyl, C _1-6 haloalkyl or -OR ⁶ , or R ⁴ and R ⁵ together with the carbon atom to which they are attached form a ring.

In some embodiments, R ⁴ is C _1-6 alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein cycloalkyl, aryl or heteroaryl is optionally substituted with one, two or three independent substituents selected from halogen or C _1-6 alkyl. In some embodiments, R ⁴ is cyclohexyl, cyclopropyl, phenyl, tetrahydropyranyl, pyridyl, pyrazolyl, thienyl, or C _1-6 alkyl, wherein cyclohexyl, cyclopropyl, phenyl, tetrahydropyranyl, pyridyl, pyrazolyl, or thienyl is optionally substituted with one, two, or three independent substituents selected from halogen or C _1-6 alkyl. In some embodiments, R ⁴ is cyclohexyl, phenyl, pyridyl, pyrazolyl, or thienyl, wherein the cyclohexyl, phenyl, pyridyl, pyrazolyl, or thienyl is optionally substituted with one, two, or three independent substituents selected from halogen or C _1-6 alkyl. In some embodiments, R ⁴ is phenyl, optionally substituted with one, two or three substituents selected from -F, -Cl or C _1-6 alkyl.

In some embodiments, R ⁵ is hydrogen, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH ₂ CH ₃ , -CH(CH ₃ ) ₂ , -CH ₂ CH ₂ CH ₂ CH ₃ , -CF ₃ or unsubstituted cyclopropyl. In some embodiments, R ⁵ is -CH ₂ CH ₃ , -CF ₃ or unsubstituted cyclopropyl.

In some embodiments, the compound is a compound of formula (I-a):

In some embodiments, the compound is a compound of formula (I-b):

In some embodiments, the compound is a compound of formula (I-c), formula (I-d), formula (I-e), formula (I-f), formula (I-g), formula (I-i), or formula (I-j):

In some embodiments, the compound is a compound of formula (I-c):

In some embodiments, the compound is a compound of formula (I-e), formula (I-k), formula (I-l), formula (I-m), formula (I-o), formula (I-p), formula (I-t), formula (I-u), or formula ( I-v):

where each of R ^b2 independently represents halogen, C _1-6 alkyl, C _1-6 haloalkyl, cyano or -OR ⁶ ; and p is 0, 1, 2 or 3.

In some embodiments, R ^a is -C _1-6 alkyl. In some embodiments, R ^a is -CH ₃ . In some embodiments, R ^a is -CH ₃ and m is 1. In some embodiments, m is 1. In some embodiments, R ^a is halogen or -C _1-6 alkyl. In some embodiments, R ^a is F or -CH ₃ . In some embodiments, R ^b is C _1-6 alkyl. In some embodiments, R ^b is -CH ₃ .

In some embodiments, the compound is a compound of formula (I-q):

In some embodiments, the compound is a compound of formula (I-r):

In some embodiments, the compound is a compound of formula (I-s):

In some embodiments, the compound is a compound of formula (I-s1) or (I-s2):

where each of R ^b2 independently represents halogen, C _1-6 alkyl, C _1-6 haloalkyl, cyano or -OR ⁶ ; and p is 0, 1, 2 or 3.

In some embodiments, R ^b is C _1-6 alkyl. In some embodiments, R ^b is -CH ₃ .

Representative examples of compounds include, but are not limited to, the compounds presented in Table 1 below.

Table 1. Typical connection examples

Connection number Structure 1-3 1-4 1-5 1-6 1-7 1-8 1-9 1-10 1-11 1-12 1-13 1-14 1-15 1-16 1-17 1-18 1-19 1-20 1-21 1-22 1-23 1-24 1-25 1-26 1-27 1-28 1-29 1-30 1-31 1-32 1-33 1-34 1-35 1-36 1-37 1-38 1-39 1-40 1-41 1-42 1-43 1-44 1-45 1-46 1-47 1-48 1-49 1-50 1-51 1-52 1-53 1-54 1-55 1-56 1-57 1-58 1-60 1-61 1-62 1-63 1-64 1-65 1-66 1-67 1-68 1-69 1-70 1-71 1-72 1-73 1-74 1-75 1-76 1-77 1-78 1-79 1-80 1-81 1-82 1-83 1-84 1-85 1-86 1-87 1-88 1-89 1-90 1-91 1-92 1-93 1-94 1-95 1-96 1-97 1-98 1-99 1-100 1-101 1-102 1-103 1-104 1-105 1-106 1-107 1-108 1-109 1-110 1-111 1-112 1-113 1-114 1-115 1-116 1-117 1-118 1-119 1-120 1-121 1-122 1-123 1-124 1-125 1-126 1-127 1-128 1-129 1-130 1-131 1-132 1-133 1-134 1-135 1-136 1-137 1-138 1-139 1-140 1-141 1-142 1-143 1-144 1-145 1-146 1-147 1-148 1-149 1-150 1-151 1-152 1-153 1-154 1-155 1-156 1-157 1-158 1-159 1-160 1-161 1-162 1-163 1-164 1-165 1-166 1-167 1-168 1-169 1-170 1-171 1-172 1-173 1-174 1-175 1-176 1-177 1-178 1-179 1-180 1-181 1-182 1-183 1-184 1-185 1-186 1-187 1-188 1-189 1-190 1-191 1-192 1-193 1-194 1-195 1-196 1-197 1-198 1-199 1-200 1-201 1-202 1-203 1-204 1-205 1-206 1-207 1-208 1-209 1-210 1-211 1-212 1-213 1-214 1-215 1-216 1-217 1-218 1-219 1-220 1-221

In some embodiments, a compound of the present invention is in the form of a salt. The compound of the present invention may be administered in the form of a free acid, a colorterion or a salt. The salt can also form a cation and a negatively charged substituent on the compound of the present invention. Suitable cationic counterions include sodium ions, potassium ions, magnesium ions, calcium ions and ammonium ions (eg, tatraalkylammonium cation such as tetramethylammonium ion). In compounds containing a positively charged substituent or basic substituent, the salt may form an anion and a positively charged substituent (eg, amino group) or basic substituent (eg, pyridyl) on the compound of the present invention. Suitable anions include chlorine, bromine, iodine, sulfate, nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate and acetate.

Pharmaceutically acceptable salts of the compounds of the present invention (for example, compounds of formula (II), formula (I), formula (I-a), formula (I-b), formula (I-c), formula (I-d), formula (I-e), formula (I-f) , formulas (I-g), formulas (I-h), formulas (I-i), formulas (I-j), formulas (I-k), formulas (I-l), formulas (I-m), formulas (I-n), formulas (I-o), formulas (I-p) , formula (I-r), formula (I-c), formula (I-s1), formula (I-s2), formula (I-t), formula (I-u), formula (I-v) or a compound shown in Table 1) also include salts derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acid addition salts include acetate, 4-acetamidobenzoate, adipate, alginate, 4-aminosalicylate, aspartate, ascorbate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, carbonate, cinnamate, cyclamate, decanoate, decanedioate, 2 ,2-dichloroacetate, digluconate, dodecyl sulfate, ethanesulfonate, ethane-1,2-disulfonate, fumarate, galactarate, glucoheptanoate, gluconate, glucoheptonate, glucoronate, glutamate, glutarate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate, at, hydrochloride , hydrobromide, hydroiodide, 1-hydroxy-2-naphthoate, 2-hydroxyethanesulfonate, isobutyrate, lactate, lactobionate, laurate, malate, maleate, malonate, mandelate, methanesulfonate, naphthalene-1,5-disulfonate, 2-naphthalene sulfonate, nicotinate, nitrate , octanoate, oleate, oxalate, 2-oxoglutarate, palmitate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, phosphonate, picrate, pivalate, propionate, pyroglutamate, salicylate, sebacate, succinate, stearate, sulfate, tartrate, thiocyanate, toluenesulfonate , tosylate, trifluoroacetate and undecanoate.

Salts derived from suitable bases include alkali metal (eg sodium), alkaline earth metal (eg magnesium), ammonium and N-(alkyl) ₄ ⁺ salts. The present invention also provides for the quaternization of any basic nitrogen-containing groups of compounds disclosed herein. With such quaternization, water-soluble or oil-dispersible compounds can be obtained.

As used herein, the compounds of the present invention, including the compounds of formula (II), are defined to include their pharmaceutically acceptable derivatives or prodrugs. The term “pharmaceutically acceptable derivative or prodrug” means any pharmaceutically acceptable salt, ester, ester salt, or other derivative of a compound of the present invention that, when administered to a recipient, is capable of forming (directly or indirectly) the compound of the present invention. Particularly preferred derivatives and prodrugs are derivatives and prodrugs that increase the bioavailability of the compounds of the present invention when such compounds are administered to a mammal (eg, allowing the orally administered compound to be more readily absorbed into the bloodstream), or that enhance delivery of the parent compound to a biological compartment (eg, the brain or lymphatic system) relative to the original species. Preferred prodrugs include derivatives in which a group is added to the structure of the formulas described in the present invention that increases their solubility in water or active transport across the intestinal membrane.

Any formula or compound of the present invention also includes both unlabeled and isotopically labeled forms of the compounds, wherein the structures of the isotopically labeled compounds are represented by the given formulas, but with the difference that one or more atoms are replaced by atom(s) of the selected atomic mass or selected atomic mass number. Examples of isotopes that can be introduced into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as ² H, ³ H, ¹¹ C, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ F ⁵¹ P, ³² P, ³⁵ S, ³⁶ Cl, ¹²⁵ I, respectively. The invention includes various isotopically labeled compounds that are defined in the present invention, for example compounds in which radioactive isotomes such as ³ H, ¹³ C, and ¹⁴ C are present. Such isotopically labeled compounds are useful in metabolic studies (with ¹⁴ C), kinetic studies reactions (eg 'H or ^3H ), detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including drug or tissue substrate assay techniques, or in radioactive treatment techniques for patients. In particular, ^{an 18} F or labeled compound may be particularly desirable for PET or SPECT studies, and isotopically labeled compounds of the present invention and prodrugs thereof can be prepared by implementing the procedures disclosed in the schemes and in the examples and preparations described below, substituting a readily available an isotopically labeled reagent is an isotopically unlabeled reagent.

In addition, substitution with heavier isotopes, particularly deuterium (ie, ² H or D), may provide certain therapeutic benefits due to greater metabolic stability, such as increased in vivo half-life, reduced dosage requirements, and improved therapeutic index. It will be understood that deuterium is considered in this context as a substitute for the compound of the formula described in the present invention. The concentration of such a heavier isotope, in particular deuterium, can be determined by the isotomic enrichment factor. The term "isotopic enrichment factor" when used herein means the ratio of the abundance of said isotope to the natural abundance of said particular isotope. If a substituent in a compound of the present invention is designated deuterium, the isotopic enrichment factor of such compound for each designated deuterium atom is at least 3500 (52.5% deuterium injected per designated deuterium atom), at least 4000 (60% deuterium injected), at least 4500 (67.5% deuterium injected), at least 5000 (75% deuterium injected), at least 5500 (82.5% deuterium injected), at least 6000 deuterium), at least 6333.3 (95% deuterium injected), at least 6466.7 (97% deuterium injected), at least 6600 (99% deuterium injected) or at least 8633.3 (99.5% deuterium injected).

The isotopically labeled compounds of the present invention can generally be prepared by standard methods known to one skilled in the art, or by methods similar to those described in the accompanying Examples and Preparation Examples, using suitable isotopically labeled reagents in place of the previously used isotopically unlabeled reagents. Pharmaceutically acceptable solvates in accordance with the present invention include solvates in which the crystallization solvent may be isotopically substituted, for example D ₂ O, D ₂ -acetone, D ₂ -DMSO.

Any asymmetric atom (eg, carbon atom or the like) of the compound(s) of the present invention may be present in a racemic or enantiomerically enriched mixture, for example (R)-, (S)- or (RS)-configuration. In some embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least at least 95% enantiomeric excess, or at least 99% enantiomeric excess in the (R) or (S) configuration. Substituents on atoms with unsubstituted bonds may, if possible, be present in cis-(Z)- or trans-(E)-form. Accordingly, the term “compound of the present invention,” when used herein, means a compound of the present invention, which may be in the form of one of possible isomers, rotamers, atropisomers, tautomers, and mixtures thereof, such as substantially pure geometric (cis or trans) isomers, diastereomers, optical isomers (antipodes), racemates and mixtures thereof. Any resulting mixtures of isomers can be separated, based on differences in the physicochemical properties of their constituent compounds, into pure or essentially pure geometric or optical isomers, diastereomers, racemates, for example, using chromatography or fractional crystallization.

Methods for synthesizing the compounds of the present invention (eg, compounds of formula (II)) may independently produce compositions comprising mixtures of two or more diastereomers of the compound. The term “diastereomeric excess” (“d.e.”) or “% diastereomeric excess” (“% d.e.”) of a composition, when used herein, refers to the excess of one diastereomer relative to another or several different diastereomers present in the composition.

In some embodiments, the term “disastereomeric excess (d.e.) of a composition”, when referring to a diastereomer of the present invention relative to another, means a diastereomeric excess of greater than 90%. In some embodiments, the term "d.e. composition" referring to a diastereomer of the present invention means a diastereomeric excess of greater than 95%. In some embodiments, the term “d.e. composition" referring to a diastereomer of the present invention means a diastereomeric excess of greater than 97%. In some embodiments, the term “d.e. composition" referring to a diastereomer of the present invention means a diastereomeric excess of greater than 99%. In some embodiments, the term “d.e. composition" referring to a diastereomer of the present invention means a diastereomeric excess of greater than 99.9%.

Any resulting racemates of the final products or intermediates can be resolved into optical antipodes by known methods, for example, separation from diastereomeric salts obtained with an optically active acid or an optically active base, and isolation of the optically active acid or base compound. In particular, a basic molecular fragment can thus be used to separate the compounds of the present invention into their optical antipodes, for example by fractional crystallization of a salt obtained with an optically active acid, for example tartaric acid, dibenzoyltartaric acid, diacetyltartaric acid, (+)-O, O ′-di-p-toluoyl-D-tartaric acid, mandelic acid, malic acid or camphor-10-sulfonic acid. Racemic products can also be separated by chiral chromatography, such as high pressure liquid chromatography (HPLC) using a chiral adsorbent.

The compounds of the present invention (eg, a compound of formula (II)) may also be present in a variety of tautomeric forms, for example, as a compound of formulas (II), (I), (I-a), (I-b), (I-c), (I-d), ( I-e), (I-f), (I-g), (I-h), (I-i), (I-j), (I-k), (I-l), (I-m), (I-n), (I-o), (I-p), (I-q) , (I-r), (I-c), (I-s1), (I-s2), (I-t), (I-u) or (I-v). In such cases, the present invention includes all tautomeric forms of the compounds of the present invention. All crystalline forms of the compounds of the present invention are included in the present invention.

A compound of the present invention (eg, a compound of formula (II)) can be tested for its ability to modulate (eg, inhibit) factor XIa or kallikrein.

Methods for synthesizing compounds

The compounds of the present invention can be synthesized by standard methods using commercially available starting materials and reagents. For example, the compounds can be synthesized using the methods disclosed in US Patent No. 7501404, or as described in the methods presented herein.

Methods of treatment, prevention and risk reduction

The compounds of the present invention (eg, compounds of formula (II)) can inhibit factor XIa or kallikrein. In some embodiments, a compound of the present invention may inhibit factor XIa and kallikrein. As a result, these compounds can be used to treat, prevent, or reduce the risk of the disorders described herein. Examples of disorders include thrombotic processes associated with coronary artery and cardiovascular disease, venous or arterial thrombosis, coagulation syndromes, ischemia (eg, coronary artery ischemia) and angina (stable and unstable), deep vein thrombosis (DVT), hepatic thrombosis venous, disseminated intravascular coagulopathy, Kasabach-Merritt syndrome, pulmonary embolism, myocardial infarction (eg, myocardial infarction with ST-segment elevation or myocardial infarction without ST-segment elevation (eg, myocardial infarction without ST-segment elevation before catheterization), cerebral infarction, cerebral thrombosis, transient ischemic attack, atrial fibrillation (eg, non-valvular atrial fibrillation), cerebral embolism, thromboembolic complications of surgery (eg, hip or knee replacement, orthopedic surgery, cardiac surgery, pulmonary surgery, abdominal organs or endarterectomy) and peripheral arterial obstruction, and may also be used to treat or prevent myocardial infarction, stroke, angina pectoris and other consequences of atheroclerotic plaque rupture. The compounds of the present invention having factor XIa or kallikrein inhibitory activity may also be used to prevent thromboembolic complications, such as venous thromboembolism, in cancer patients, including patients receiving chemotherapy and/or patients with elevated lactate dehydrogenase (LDH) levels, and to prevent thromboembolic complications during or after repair with tissue plasminogen activator or mechanical restoration of blood vessel patency. The compounds of the present invention having inhibitory activity against factor XIa or kallikrein can also be used as blood coagulation inhibitors, for example in the collection, storage and fractionation of whole blood. In addition, the compounds of the present invention may be used in acute hospitalization or periprocedural settings where the patient is at risk for a thromboembolic disorder or complication, as well as in patients who are in a hypercoagulable state, such as cancer patients.

Inhibition of factor XIa according to the present invention may be a more effective and safe method of inhibiting thrombosis compared to inhibition of other serine coagulation proteases such as thrombin or factor Xa. Administration of a small molecule factor XIa inhibitor should exhibit an effect of inhibiting thrombin generation and clot formation with no or no effect on bleeding time and little or no effect on bleeding time and little or no effect on homeostasis. These results differ significantly from those shown by other “direct acting” protease inhibitors (eg, thrombin active site inhibitors and factor Xa), which show increased bleeding time and less separation between antithrombotic efficacy and increased bleeding time. A preferred method of the present invention involves administering to a mammal a pharmaceutical composition comprising at least one compound of the present invention

Compounds of the present invention (for example, a compound of formula (II)), in particular compounds of formulas (II), (I), (I-a), (I-b), (I-c), (I-d), (I-e), (I-f), (I-g), (I-h), (I-i), (I-j), (I-k), (I-l), (I-m), (I-n), (I-o), (I-p), (I-q), (I-r), (I -c), (I-s1), (I-s2), (I-t), (I-u) or (I-v), may inhibit kallikrein. Thus, these compounds can be used for the treatment, prevention or reduction of the risk of diseases associated with inflammation, such as edema (eg, cerebral edema, macular edema and angioedema (eg, hereditary angioedema)). In some embodiments, the compounds of the present invention may be used to treat or prevent hereditary angioedema. The compounds of the present invention (eg, compounds of formula (II)) can also be used to treat, prevent or reduce the risk of, for example, stroke, ischemia (eg, coronary artery disease) and perioperative blood loss, for example, a compound of formula (II), such as compound of formulas (I), (I-a), (I-b), (I-c), (I-d), (I-e), (I-f), (I-g), (I-h), (I-i), (I-j), (I-k), (I-l), (I-m), (I-n), (I-o), (I-p), (I-q), (I-r), (I-c), (I-s1), (I-s2), (I-t), (I-u) or (I-v). The methods of the present invention can be used to treat or prevent conditions associated with factor XIa or kallikrein. Accordingly, the methods of the present invention can be used to treat the consequences of atherosclerotic plaque rupture, including cardiovascular diseases associated with activation of the coagulation cascade in thrombotic or thrombophilic conditions.

More specifically, the methods of the present invention can be used to treat, prevent, or reduce the risk of acute coronary syndromes such as coronary artery disease, myocardial infarction, unstable angina (including progressive angina), ischemia (eg, ischemia due to vascular occlusion), and cerebral infarction. The methods of the present invention may additionally be used to treat, prevent, or reduce the risk of stroke and related cerebrovascular diseases (including cerebrovascular accidents, vascular dementia, and transient ischemic attack); venous thrombosis and thromboembolism such as deep vein thrombosis (DVT) and pulmonary embolism; thrombosis associated with atrial fibrillation, ventricular hypertrophy, widespread cardiomyopathy, or heart failure; peripheral arterial disease and intermittent claudication; formation of atherosclerotic plaques and atherosclerosis of the graft; restenosis after arterial injury, induced endogenously (by rupture of an atorosclerotic plaque) or exogenously (by invasive cardiac procedures, such as damage to the vessel wall as a result of angioplasty or postcranial arterial stenting); disseminated intravascular coagulopathy, Kasabach-Merritt syndrome, cerebral thrombosis and cerebral embolism.

In addition, the methods of the present invention can be used to treat, prevent (eg, prevent), or reduce the risk of thromboembolic consequences or complications associated with cancer, surgery (eg, hip replacement, orthopedic surgery), endarterectomy, insertion of artificial heart valves , peripheral vascular surgery (eg, limbs), cardiovascular surgery, large-bore aneurysm surgery, vascular grafts, mechanical organs and implantation (eg, transcatheter aortic valve implantation) or organ transplantation (eg, liver transplantation) , tissue or cells); percutaneous coronary interventions; catheter ablation; hemophilia therapy; hemodialysis; drugs (such as tissue plasminogen activator or similar drugs and drugs for the surgical restoration of blood vessel patency) in patients with myocardial infarction, stroke, pulmonary embolism, etc. states; drugs (such as oral contraceptives, hormone replacement therapy and hepatin, for example for the treatment of heparin-induced thrombocytopeia); sepsis (such as sepsis associated with disseminated intravascular coagulation syndrome); pregnancy or childbirth; and other chronic medical conditions. The methods of the present invention can be used to treat thrombosis due to restriction of movement (eg, immobilization, hospitalization, bed rest or limb immobilization, eg with immobilization bandages, etc.).

In addition, the compounds of the present invention (for example, a compound of formula (II)) or pharmaceutically acceptable salts or compositions thereof can be used for the treatment, prevention and reduction of the risk of thromboembolic complications, such as venous thromboembolism, deep vein thrombosis or pulmonary embolism, or associated complications in the patient where the patient is exposed to the artificial surface. The artificial surface may come into contact with the patient's blood, for example as an extracorporeal surface or the surface of an implantable device. Such artificial surfaces include, but are not limited to, the surfaces of dialysis catheters, cardiopulmonary bypass circuits, artificial heart valves, such as mechanical heart valves (MHV), ventricular assist devices, small grafts, central venous catheters, extracorporeal membrane oxygenation (ECMO) machines. In addition, a thromboembolic disorder or thromboembolic complication may be caused by or associated with the artificial surface. For example, foreign surfaces and various components of mechanical heart valves (MHVs) are prothrombotic and promote thrombin generation through the intrinsic metabolic pathway of blood coagulation. In addition, thrombin and FXa inhibitors are contraindicated in thromboembolic disorders or related complications caused by artificial surfaces, such as modified hemostatic valve (MHV) surfaces, because these inhibitors are ineffective at blocking the intrinsic metabolic pathway at plasma levels that will not cause significant bleeding . Thus, the compounds of the present invention, which can be used, for example, as inhibitors of factor XIa, are considered as alternative therapeutic agents for these purposes.

The compounds of the present invention (eg, a compound of formula (II)) or pharmaceutically acceptable salts or compositions thereof may also be used to treat, prevent, or reduce the risk of atrial fibrillation in a patient in need thereof. For example, a patient may be at high risk of developing atrial fibrillation. The patient may also need dialysis, such as kidney dialysis. The compounds of the present invention (eg, a compound of formula (II)) or pharmaceutically acceptable salts or compositions thereof can be administered before, during or after dialysis. Direct oral coagulants (DOACs) available today, such as some FXa or thrombin inhibitors, are contraindicated for atrial fibrillation in this condition. Therefore, the compounds of the present invention, which can be used, for example, as inhibitors of factor XIa, are considered as alternative therapeutic agents for these purposes. In addition, the patient may be at high risk of bleeding. In some embodiments, the patient may suffer from end-stage renal disease. In other cases, the patient does not need dialysis, such as kidney dialysis. In addition, atrial fibrillation may be associated with another thromboembolic complication such as a blood clot.

Further, the compounds of the present invention (eg, a compound of formula (II)) or pharmaceutically acceptable salts or compositions thereof can be used for the treatment, prevention or reduction of the risk of hypertension, eg arterial hypertension, in a patient. In some embodiments, hypertension, such as arterial hypertension, can lead to atherosclerosis. In some embodiments, the hypertension may be pulmonary arterial hypertension.

Moreover, the compounds of the present invention (for example, a compound of formula (II)) or pharmaceutically acceptable salts or compositions thereof can be used for the treatment, prevention or reduction of the risk of disorders such as heparin-induced thrombocytopenia, heparin-induced thrombocytopenia and thrombosis or thrombotic microangiopathy , such as hemolytic uremic syndrome (HUS) or thrombotic thrombocytopenic purpura (TTP).

The compounds of the present invention (eg, a compound of formula (II)) or pharmaceutically acceptable salts or compositions thereof can be used to reduce inflammation in a patient. In some embodiments, the inflammation may be vascular inflammation. In some embodiments, vascular inflammation may be accompanied by atherosclerosis. In some embodiments, vascular inflammation may be accompanied by thrombotic disease in a patient. In some embodiments, the vascular inflammation may be angiotensin II-induced vascular inflammation.

The compounds of the present invention (for example, a compound of formula (II)) or pharmaceutically acceptable salts or compositions thereof can be used for the treatment, prevention or reduction of the risk of renal dysfunction or renal dysfunction, including end-stage renal failure, renal dysfunction associated with hypertension in a patient, renal fibrosis and renal failure.

The methods of the present invention may also be used to maintain patency of blood vessels, for example in patients undergoing transluminal coronary angioplasty, or in connection with vascular surgery such as bypass surgery, arterial reconstruction, atherectomy, vascular grafts, stent reconstruction, as well as implantation and transplantation of organs, tissues or cells. The methods of the present invention can be used to inhibit blood coagulation during the collection, storage, fractionation, and use of whole blood. For example, the methods of the present invention can be used to maintain whole and fractionated blood in the liquid phase as needed for analytical and biological testing, such as ex vivo studies of platelet and other cell function, bioanalytical techniques and quantitative analysis of blood components, or for the maintenance of extracorporeal circuits. , as in renal replacement solution (eg, hemodialysis) or during surgery (eg, open heart surgery, such as coronary artery bypass grafting). In some embodiments, the renal replacement solution may be used to treat patients with acute renal failure. In some embodiments, the renal replacement solution may be continuous renal replacement therapy.

In addition, the methods of the present invention can be used to treat and prevent prothrombotic complications of cancer. The methods may be used to treat tumor growth as an adjunct to chemotherapy, to prevent angiogenesis, and to treat cancer, particularly lung cancer, prostate cancer, colon cancer, breast cancer, ovarian cancer, and bone cancer.

In one aspect, the present invention relates to a method of reducing the risk of stroke in a patient who has suffered an ischemic attack, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition, comprising a compound of formula (II). In some embodiments, administration reduces the patient's risk of stroke compared to a patient not administered the compound.

In one aspect, the present invention provides a method of reducing the risk of stroke or systemic embolism in a patient suffering from atrial fibrillation, comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention, e.g. a composition comprising a compound of formula (II).

In another aspect, the present invention relates to a method of reducing the risk of systemic embolism outside the CNS in a patient who has suffered an ischemic attack, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention , for example a composition comprising a compound of formula (II). In some embodiments, administration reduces the risk of systemic embolism outside the CNS in a patient compared to a patient who is not administered the compound.

In one aspect, the present invention relates to a method of treating deep vein thrombosis, comprising administering to a patient who has suffered an ischemic attack an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition, comprising a compound of formula (II).

In one aspect, the present invention provides a method of reducing the risk of recurrent deep vein thrombosis, comprising administering to a patient who has experienced deep vein thrombosis an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention , for example a composition comprising a compound of formula (II). In some embodiments, administration reduces the patient's risk of recurrent deep vein thrombosis compared to a patient not receiving the compound.

In one aspect, the present invention provides a method for preventing deep vein thrombosis in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula ( II). In some embodiments, the patient undergoes orthopedic surgery or knee or hip replacement surgery.

In one aspect, the present invention relates to a method of reducing the risk of recurrent pulmonary embolism, comprising administering to a patient who is suffering from a pulmonary embolism an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention, e.g. a composition comprising a compound of formula (II).

In one aspect, the present invention relates to a method of reducing the risk of recurrent pulmonary embolism, comprising administering to a patient who has suffered from deep vein thrombosis an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II). In some embodiments, administration reduces the patient's risk of recurrent pulmonary embolism compared to a patient not administered the compound.

In one aspect, the present invention relates to a method of treating pulmonary embolism in a patient who has suffered from deep vein thrombosis, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention, for example a composition comprising a compound of formula (II).

In one aspect, the present invention provides a method for preventing pulmonary embolism in a patient who has suffered from pulmonary embolism, comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, e.g. comprising a compound of formula (II).

In one aspect, the present invention provides a method for preventing pulmonary embolism in a patient who has suffered from pulmonary embolism, comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, e.g. comprising a compound of formula (II).

In one aspect, the present invention relates to a method of preventing pulmonary embolism in a patient who has suffered from deep vein thrombosis, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention, for example a composition comprising a compound of formula (II).

In one aspect, the present invention relates to a method of treating a patient who has suffered an ischemic attack, comprising administering a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II) , to the patient within 12 hours or less, 10 hours or less, 9 hours or less, 8 hours or less, 7 hours or less, 6 hours or less, or 1 hour or less, after the onset of the patient's ischemic attack.

In one aspect, the present invention relates to a method of treating deep vein thrombosis in a patient who has previously been administered an anticoagulant, comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, e.g. comprising a compound of formula (II).

In one aspect, the present invention relates to a method of treating pulmonary embolism in a patient who has previously been administered an anticoagulant, comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, e.g. comprising a compound of formula (II). In some embodiments, the anticoagulant is administered parenterally over a period of 5-10 days.

In one aspect, the present invention relates to a method of treating a patient who has suffered an ischemic attack, comprising: administering a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II ), the patient in the range of more than 2 hours to 12 hours, for example from more than 2 hours to 10 hours or less, from more than 2 hours to 8 hours or less, after the onset of an ischemic attack in the patient.

In one aspect, the present invention relates to a method of treating acute coronary syndrome in a patient, comprising administering to a patient in need thereof a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising compound of formula (II).

In one aspect, the present invention provides a method of inhibiting factor XIa in a patient, comprising administering to a patient who has suffered ischemia an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition, comprising a compound of formula (II). In some embodiments, the ischemia is coronary ischemia.

In one aspect, the present invention relates to a method of treating a patient with edema, comprising administering a compound of the present invention, eg a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, eg a composition comprising a compound of formula (II).

In one aspect, the present invention provides a method of inhibiting kallikrein in a patient, comprising administering to a patient with edema an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula ( II).

In one aspect, the present invention provides a method of treating a thromboembolic consequence or thromboembolic complication in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising compound of formula (II). In some embodiments, the thromboembolic consequence or thromboembolic complication is associated with intervention in peripheral blood vessels (eg, extremities), hemodialysis, catheter ablation, cerebrovascular intervention, organ transplantation (eg, liver), surgery (eg, orthopedic surgery, pulmonary surgery, abdominal or cardiac surgery (eg, open heart surgery), transcatheter aortic valve implantation, large-bore procedure used to treat an aneurysm, percutaneous coronary intervention, or hemophilia therapy. In some embodiments, the surgery is orthopedic surgery, pulmonary surgery, abdominal surgery, or cardiac surgery.

In one aspect, the present invention relates to a method of treating restenosis following arterial injury in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II). In some embodiments, arterial injury occurs following cranial artery stenting.

In one aspect, the present invention relates to a method of treating hepatic vascular thrombosis in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula ( II).

In one aspect, the present invention provides a method of treating non-ST segment elevation myocardial infarction or ST segment elevation myocardial infarction, comprising administering to a patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition according to the present invention, for example a composition comprising a compound of formula (II).

In one aspect, the present invention relates to a method of maintaining blood vessel patency, comprising administering to a patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II) . In some embodiments, the patient is suffering from acute renal failure. In some embodiments, the patient additionally receives continuous renal replacement therapy.

In one aspect, the present invention relates to a method of treating hypertension in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II) . In some embodiments, the hypertension is arterial hypertension.

In one aspect, the present invention provides a method of reducing the risk of hypertension in a patient, comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, e.g., a composition comprising a compound of formula (II) ). In some embodiments, the hypertension is arterial hypertension.

In one aspect, the present invention provides a method for preventing hypertension in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II) . In some embodiments, the hypertension is arterial hypertension.

In one aspect, the present invention provides a method of reducing inflammation in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II) . In some embodiments, the inflammation is vascular inflammation. In some embodiments, the vascular inflammation is angiotensin II-induced vascular inflammation.

In one aspect, the present invention provides a method of preventing vascular infiltration of leukocytes in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula ( II).

In one aspect, the present invention provides a method of preventing endothelial dysfunction induced by angiotensin II in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising compound of formula (II).

In one aspect, the present invention provides a method of preventing excessive thrombin production in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula ( II).

In one aspect, the present invention relates to a method of treating renal dysfunction associated with hypertension in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising compound of formula (II).

In one aspect, the present invention relates to a method of preventing renal dysfunction associated with hypertension in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising compound of formula (II).

In one aspect, the present invention provides a method of reducing a patient's risk of renal dysfunction associated with hypertension, comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, e.g., a composition comprising a compound of formula (II).

In one aspect, the present invention relates to a method of treating renal dysfunction associated with hypertension in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising compound of formula (II).

In one aspect, the present invention provides a method of preventing renal dysfunction associated with hypertension in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising compound of formula (II).

In one aspect, the present invention provides a method of reducing a patient's risk of renal dysfunction associated with hypertension, comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, e.g., a composition comprising a compound of formula (II).

In one aspect, the present invention relates to a method of treating renal fibrosis in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II ).

In one aspect, the present invention provides a method for preventing renal fibrosis in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II) ).

In one aspect, the present invention provides a method of reducing the risk of renal fibrosis in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula ( II).

In one aspect, the present invention provides a method of treating renal failure in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II) ).

In one aspect, the present invention provides a method for preventing renal failure in a patient, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II) ).

In one aspect, the present invention provides a method of reducing the risk of renal failure in a patient, comprising administering to the patient an effective amount of a compound of the present invention, such as a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, such as a composition comprising a compound of formula ( II).

In one aspect, the present invention relates to a method of treating a thromboembolic complication in a patient in need thereof, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II), where the patient is exposed to an artificial surface.

In one aspect, the present invention relates to a method of reducing the risk of a thromboembolic complication in a patient in need thereof, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition, comprising a compound of formula (II), wherein the patient is exposed to an artificial surface.

In one aspect, the present invention relates to a method of preventing a thromboembolic complication in a patient in need thereof, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II), where the patient is exposed to an artificial surface.

In one aspect, the present invention provides a method of treating heparin-induced thrombocytopenia in a patient in need thereof, comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, e.g. comprising a compound of formula (II).

In one aspect, the present invention provides a method of reducing the risk of heparin-induced thrombocytopenia in a patient in need thereof, comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention, e.g. a composition comprising a compound of formula (II).

In one aspect, the present invention provides a method for preventing heparin-induced thrombocytopenia in a patient in need thereof, comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, e.g. comprising a compound of formula (II).

In one aspect, the present invention relates to a method of treating heparin-induced thrombocytopenia and thrombosis in a patient in need thereof, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention, for example a composition comprising a compound of formula (II).

In one aspect, the present invention provides a method of reducing the risk of heparin-induced thrombocytopenia and thrombosis in a patient in need thereof, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention , for example a composition comprising a compound of formula (II).

In one aspect, the present invention provides a method for preventing heparin-induced thrombocytopenia and thrombosis in a patient in need thereof, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention, for example a composition comprising a compound of formula (II).

In one aspect, the present invention relates to a method of preventing a thromboembolic complication in a patient in need thereof, comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, for example a composition comprising a compound of formula (II), wherein the patient suffers from cancer.

In one aspect, the present invention provides a method of treating thrombotic microangiopathy in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of formula (II) of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof invention, for example a composition comprising a compound of formula (II).

In one aspect, the present invention provides a method of reducing the risk of thrombotic microangiopathy in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention, for example a compound of formula (II) or a pharmaceutically acceptable salt or composition thereof of the present invention, e.g. a composition comprising a compound of formula (II).

In another aspect, the present invention provides a method for preventing thrombotic microangiopathy in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of the present invention, e.g., a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a composition of the present invention, e.g. comprising a compound of formula (II).

In some embodiments, the patient is a patient with non-valvular atrial fibrillation. In some embodiments, the patient has one or more of the following risk factors: a history of stroke, transient ischemic attack, or systemic embolism outside the CNS. In some embodiments, the patient has one or more of the following risk factors: age 75 years or older, hypertension, heart failure or left ventricular ejection fraction, or diabetes mellitus.

Extracorporeal membrane oxygenation (ECMO)

The compounds of the present invention may be used to treat, prevent, or reduce the risk of a thromboembolic complication in a patient in need thereof where the patient is exposed to an artificial surface such as an extracorporeal membrane oxygenation (ECMO) machine (vide supra), which may be used as salvage therapy with heart or pulmonary failure. The surface of the ECMO device that is in direct contact with the patient may be a prothrombotic surface, which may cause a thromboembolic complication such as venous thromboembolism, such as deep vein thrombosis or pulmonary embolism, leading to difficulties in treating the patient requiring ECMO. Blood clots in the circuit are the most common mechanical complication (19%). Large blood clots can cause oxygenator malfunction and lead to pulmonary or systemic embolism.

ECMO is often administered with a continuous infusion of heparin as an anticoagulant to prevent blood clots. However, insertion of a cannula may cause damage to the internal jugular vein, causing severe internal bleeding. Bleeding occurs in 30-40% of patients receiving ECMO and can be life-threatening. This severe bleeding is due to both the need for continuous heparin infusion and platelet dysfunction. Approximately 50% of reported deaths are due to severe bleeding complications. Aubron et al (Critical Care 2013, 17:R73) analyzed factors associated with ECMO outcomes.

The compounds of the present invention, which can be used as, for example, factor XIa inhibitors, are considered as an alternative replacement for Hepatir in ECMO therapy. The compounds of the present invention are considered to be effective drugs for blocking the intrinsic metabolic pathway at plasma levels that do not cause severe bleeding.

Ischemia

The term "ischemia" or "ischemic attack" means a vascular disease, usually involving vascular occlusion or restriction of blood supply to tissue. Ischemia can cause a lack of oxygen and glucose needed for cellular metabolism. Ischemia is usually caused by problematic blood vessels, leading to tissue damage or dysfunction. Ischemia may also be associated with local loss of blood or oxygen in a given part of the body due to congestion (eg, vasoconstriction or embolism). Causes include embolism, atherosclerotic artery thrombosis, trauma, venous problems, aneurysm, cardiac disease (eg, myocardial infarction, mitral valve disease, chronic atrial fibrillation, cardiomyopathy, and prosthetics), trauma or traumatic injury (eg, injury to a limb causing partial or complete vascular occlusion), thoracic outlet syndrome, atherosclerosis, hypoglycemia, tachycardia, hypotension, extrinsic compression of a blood vessel (eg, tumor), sickle cell anemia, localized severe cold (eg, frostbite), tourniquet application, glutamate receptor stimulation, arteriovenous malformations, rupture of important blood vessels supplying a tissue or organ, and anemia.

A transient ischemic attack generally refers to a transient (eg, short-lived) episode of neurological dysfunction caused by loss of blood flow (eg, to the brain, spinal cord, or retina) without acute infarction (eg, tissue death). In some embodiments, the transient ischemic attack lasts for less than 72 hours, 48 hours, 24 hours, 12 hours, 10 hours, 8 hours, 4 hours, 2 hours, 1 hour, 45 minutes, 30 minutes, 20 minutes, 15 minutes, 10 minutes, 5 minutes, 4 minutes, 3 minutes, 2 minutes or 1 minute.

Angioedema

Angioedema is a rapid swelling of the dermis, subcutaneous tissue, mucous membrane and submucosal tissues. Angioedema is usually classified as either hereditary or acquired.

"Acquired angioedema" may be immunological, non-immunological, or idiopathic; which is caused, for example, by allergies as a side effect of medications, such as ACE inhibitors.

The term "hereditary angioedema" or "HAO" refers to a generic disorder that intermittently results in acute swelling (eg, swelling) that can occur in almost all parts of the body, including the face, limbs, neck, throat, larynx, hands, and feet. , gastrointestinal tract and genitals. Attacks of HAE can often be life-threatening, their severity depending on the area affected, for example attacks in the abdominal region can lead to intestinal obstruction, while swelling of the larynx and upper respiratory tract can lead to asphyxia. The pathogenesis of hereditary angioedema may be associated with unopposed activation of the contact metabolic pathway through the initial production of kallikrein or coagulation factors (eg, factor XII).

Objective signs and symptoms of the disease include swelling (edema), for example, of the face, mucous membranes of the mouth or throat and tongue. Itching, pain, decreased sensation in the affected areas, urtricaria (i.e., urticaria), or airway stridor may also be objective signs of angioedema. However, there may not be any associated itching or urticaria, such as with hereditary angioedema. Patients with HAE may experience abdominal pain (eg, abdominal pain lasting for one to five days, attacks of abdominal pain with an increase in the patient's white blood cell count), vomiting, weakness, watery diarrhea, or rash.

Bradykinin plays an important role in angioedema, especially hereditary angioedema. Bradykinin is released by various cell types in response to a number of different stimuli and is a pain mediator. Interference with bradykinin production or degradation can lead to angioedema. In hereditary angioedema, continuous production of the enzyme kallikrein may contribute to the formation of bradykinin. Inhibition of kallikrein may interfere with bradykinin production and treat or prevent angioedema.

Pharmaceutical compositions

The compositions of the present invention include a compound of the present invention (for example, a compound of formula (II), for example a compound of formulas (I), (I-a), (I-b), (I-c), (I-d), (I-e), (I-f), ( I-g), (I-h), (I-i), (I-j), (I-k), (I-l), (I-m), (I-n), (I-o), (I-p), (I-q), (I-r), (I- c), (I-s1), (I-s2), (I-t), (I-u) or (I-v), as well as additional therapeutic agents, if present, in amounts effective to provide treatment of the disease or symptoms of the disease (eg , such as diseases associated with factor Ia or kallikrein).

Pharmaceutically acceptable carriers, adjuvants and excipients that may be used in the pharmaceutical composition of the present invention include, but are not limited to, ion exchange resins, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS). ), such as d-α-tocopherol polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tweens or other similar polymer delivery matrices, serum proteins such as human serum albumin, buffer systems such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinylpyrrolidone, substances on cellulose based, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, block copolymers of polyethylene and polyoxypropylene, polyethylene glycol and lanolin. Cyclodextrins, such as α-, β-, and γ-cyclodextrin or chemically modified derivatives thereof, such as hydroxyalkylcyclodextrins, including 2- and 3-hydroxypropyl-β-cyclodextrins or other soluble derivatives, can also be used successfully to improve the delivery of compounds of the formulas the present invention.

Methods of administration

The pharmaceutical compositions of the present invention can be administered orally, rectally, or parenterally (eg, intravenous infusion, intravenous bolus injection, inhalation, implantation). The term “parenteral” when used herein includes subcutaneous, intradermal, intravenous (eg, intravenous infusion, intravenous bolus injection), nasal, inhalation, pulmonary, transdermal, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial injection or other infusion methods. The pharmaceutical compositions of the present invention may contain any standard non-toxic pharmaceutically acceptable carriers, adjuvants or excipients. In some cases, the pH of the drug may be adjusted using pharmaceutically acceptable acids, bases, or buffers to increase the stability of the compound in the drug or its delivery form.

The pharmaceutical compositions may be presented in the form of a sterile injectable preparation, for example, in the form of a sterile aqueous or oily solution or a sterile aqueous or oily injectable suspension. This suspension can be prepared in accordance with methods known in the art, using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example a solution in 1,3-butanediol. Acceptable diluents and solvents that can be used are mannitol, water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils have been successfully used as solvents or suspending media. Any light, fixed oil can be used for this purpose, including synthetic mono- or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives can be used to prepare injectable preparations, as can natural pharmaceutically acceptable oils such as olive oil or castor oil, especially their polyoxyethylated forms. These oily solutions or suspensions may also contain a long-chain alcohol diluent or dispersant or carboxymethylcellulose or similar additives that are commonly used in pharmaceutically acceptable dosage form preparations such as emulsions and/or suspensions. Other conventionally used surfactants such as Tweens or Spans or other similar emulsifiers or bioavailability additives that are conventionally used in the manufacture of pharmaceutically acceptable solid, liquid or other dosage forms may also be used to formulate the formulation.

The pharmaceutical compositions of the present invention can be administered orally in any orally acceptable dosage form, including, but not limited to, capsules, tablets, emulsions and aqueous suspensions, dispersions and solutions. In the case of oral tablets, carriers commonly used include lactose and corn starch. Lubricants such as magnesium stearate are also commonly added. For oral administration in capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions or emulsions are administered orally, the active ingredient may be suspended or dissolved in the oil phase in combination with emulsifiers or suspending agents. Certain sweeteners, flavorings, colors, or flavor masking additives may be added if desired.

The compounds of the present invention can be administered, for example, by intravenous injection (eg, intravenous infusion, intravenous bolus injection), intra-arterial, subcutaneous, intraperitoneal, intramuscular, or subcutaneous; or orally, buccally, nasally, transmucosally, topically at a dose ranging from about 0.5 to about 100 mg/kg body weight, alternatively at doses ranging from 1 mg to 1000 mg/dose, every 4 to 120 hours, or in accordance with the requirements of a particular medicinal product. The methods described in the present invention involve the administration of an effective amount of a compound or composition of a compound to provide the desired or claimed effect. Typically, the pharmaceutical compositions of the present invention will be administered from about 1 to about 6 times per day (eg, by intravenous bolus injection) or, alternatively, by continuous infusion. Such administration may be used as long-term or short-term therapy. The amount of active ingredient that can be combined with the carrier to produce a unit dose dosage form will vary depending on the host being treated and the particular route of administration. A typical formulation will contain from about 5% to about 95% active compound (w/w). Alternatively, such preparations contain from about 20% to about 80% active compound.

In some embodiments, pharmaceutical compositions formulated for oral administration or intravenous administration are administered to a patient from 1 time per day to 6 times per day (eg, 2 times a day or 4 times a day). In some embodiments, the pharmaceutical composition formulated for oral administration is administered to a patient 1 time per day to 6 times per day (eg, 2 times a day or 4 times a day) for about 3 to 9 months. In some embodiments, the pharmaceutical composition formulated for oral administration is administered to a patient between 1 time per day and 6 times per day (eg, 2 times a day or 4 times a day) for the rest of his or her life.

In some embodiments, the compound is administered orally or parenterally. In some embodiments, the compound is administered orally. In some embodiments, the compound is administered parenterally. In some embodiments, the compound or a pharmaceutically acceptable salt or pharmaceutical composition thereof is administered after the patient has stopped using a direct oral anticoagulant. In some embodiments, the patient has used an oral anticoagulant. In some embodiments, the patient is a mammal. In some embodiments, the mammal is a human.

Combinations

In carrying out the methods of the present invention, it may be desirable to administer the compounds of the present invention (eg, factor XIa or kallikrein inhibitors) in combination with each other and one or more other drugs to achieve therapeutic effect, such as antithrombotic or anticoagulant agents, antihypertensive agents, anti-ischemic drugs, anti-arrhythmic drugs, platelet function inhibitors, etc. For example, the methods of the present invention are carried out by administering small molecule inhibitors of factor XIa or kallikrein in combination with a small molecule inhibitor of factor XIa or kallikrein. More specifically, the methods of the present invention can be carried out by administering small molecule inhibitors of factor XIa or kallikrein in combination with aspirin, clopidogrel, ticlopidine or CS-747, warfarin, low molecular weight heparins (such as LOVENOX), GPIIb/GPIIIa blockers, PAI-1 inhibitors such as XR-330 and T-686, P2Y1 and P2Y12 receptor antagonists; thromboxane receptor antagonists (such as ifetroban), prostacyclin mimetics, thromboxane A synthetase inhibitors (such as picotamide), serotonin-2 receptor antagonists (such as ketanserin); compounds that inhibit other coagulation factors such as FVII, FVIII, FIX, FX, prothrombin, TAFI and fibrinogen, or other compounds that inhibit FXI or kallikrein; fibrinolytics such as TPA, streptokinase, PAI-1 inhibitors, and α-2-antiplasmin inhibitors such as anti-α-2-antiplasmin antibodies, fibrinogen receptor antagonists, α-1-antitrypsin inhibitors, lipid-lowering agents such as HMG- CoA reductase (eg, pravastatin, simvastatin, atorvastatin, fluvastatin, cerivastatin, AZ4522 and itavastatin), and microsomal triglyceride transfer protein inhibitors (such as those disclosed in US Patent Nos. 5,739,135, 5,712,279 and 5,760,246); antihypertensive drugs such as angiotensin-converting enzyme inhibitors (eg captopril, lisinopril or fosinopril); angiotensin II receptor antagonists (for example, irbesartan, losartan or valsartan); ACE/NEP inhibitors (eg, omapatrilate and hemopatrilate); or β-blockers (such as propranolol, nadolol and carvedilol). The methods of the present invention can be carried out by administering small molecule inhibitors of factor XIa or kalikrein in combination with antiarrhythmic agents, such as drugs for the treatment of atrial fibrillation, for example amiodarone or dofetilide. The methods of the present invention may also be administered in combination with continuous renal replacement therapy to treat, for example, acute renal failure.

When carrying out the methods of the present invention, it may be desirable to administer the compounds of the present invention (factor XIa or kallikrein inhibitors) in combination with drugs that increase the levels of cAMP or cGMP in cells to obtain a therapeutic benefit. For example, the compounds of the present invention may be beneficial when used in combination with phosphodiesterase inhibitors, including PDE1 inhibitors (such as those described in the Journal of Medicinal Chemistry, Vol. 40, pp. 2196-2210 [1997]), PDE2 inhibitors, PDE3 inhibitors (such as revizinone, pimobendan or olprinone), PDE4 inhibitors (such as rolipram, cilomilast or piclamilast), PDE7 inhibitors or other PDE inhibitors such as dipyridamole, cilostazol, sildenafil, denbutyline, theophylline (1,2-dimethylxanthine), ARIFLOT .TM. (i.e., cis-4-cyano-4-[3-(cyclopentyloxy)-4-methoxyphenyl]cyclohexane-1-carboxylic acid), arophylline, roflumilast, C-11294A, CDC-801, BAY-19-8004 , cipamphylline, SCH351591, YM-976, PD-189659, mesiopram, pumafentrine, CDC-998, IC-485 and KW-4490.

The methods of the present invention can be carried out by administering the compounds of the present invention in combination with prothrombin drugs, such as tissue plasminogen activator (natural or recombinant), streptokinase, reteplase, activase, lanoteplase, urokinase, prourokinase, ASPAC (anisolated streptokinase plasminogen activator complex), plasminogen activators of animal salivary glands, etc. The methods of the present invention can be carried out by administering the compounds of the present invention in combination with β-adrenergic agonists such as albuterol, terbutaline, formoterol, salmeterol, bitolterol, pilbuterol or fenoterol; aanticholinergics such as ipratropium bromide; anti-inflammatory corticosteroids such as beclomethasone, triamcinolone, budesonide, fluticasone, flunisolide or dexamethasone; and anti-inflammatory drugs such as cromolyn, nedocromil, theophylline, zileuron, zafirlukast, monteleukast and pranleukast. Small molecule inhibitors of factor XIa or kallikrein may act synergistically with one or more of the above agents. Thus, reduced doses of the thrombolytic agent(s) can be used, obtaining the beneficial effects of administering the compounds while minimizing possible hemorrhagic and other side effects.

A course of treatment

The compositions of the present invention include an effective amount of a compound of the present invention (eg, a factor XIa inhibitor or kallikrein) in combination with one or more other drugs (eg, an additional therapeutic agent), such as antithrombotic or anticoagulant agents, antihypertensive agents, antiischemic agents, antiarrhythmics, platelet function inhibitors, etc., to achieve therapeutic benefits.

In some embodiments, the additional therapeutic agent is administered after administration of a compound of the present invention (eg, a factor XIa inhibitor or kallikrein). In some embodiments, the additional therapeutic agent is administered at 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 14 hours, 18 hours, 24 hours, 48 hours, 72 hours. or a longer period of time after administration of a compound of the present invention (eg, a factor XIa inhibitor or kallikrein). In some embodiments, the additional therapeutic agent is administered (eg, orally) upon discharge from a healthcare facility (eg, hospital).

In some embodiments, a compound of the present invention (eg, a factor XIa or kallikrein inhibitor) and an additional therapeutic agent are formulated into a single composition or dosage form. In some embodiments, a compound of the present invention (eg, a factor XIa or kallikrein inhibitor) and the additional therapeutic agent are administered separately. In some embodiments, a compound of the present invention (eg, a factor XIa or kallikrein inhibitor) and the additional therapeutic agent are administered sequentially. In some embodiments, a compound of the present invention (eg, a factor XIa or kallikrein inhibitor) and the additional therapeutic agent are administered separately and sequentially. Typically, at least one compound of the present invention (e.g., a factor XIa or kallikrein inhibitor) and the additional therapeutic agent are administered parenterally (e.g., nasally, buccally, inhalation, implantation, transdermally, intravenously (e.g., intravenous infusion, intravenous bolus injection) , subcutaneous, intradermal, intranasal, pulmonary, percutaneous, intra-articular, intra-arterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or other infusion methods); orally; or rectally, eg by intramuscular injection or intravenously (eg, intravenous infusion, intravenous bolus injection)). In some embodiments, a compound of the present invention is administered parenterally (eg, nasally, buccally, intravenously (eg, intravenous infusion, intravenous bolus injection), or intramuscularly). In some embodiments, the additional therapeutic agent is administered orally. In some embodiments, a compound of the present invention (e.g., a factor XIa or kallikrein inhibitor) is administered parenterally (e.g., nasally, buccally, intravenously (e.g., intravenous infusion, intravenous bolus injection, or intramuscular injection) and the additional therapeutic agent is administered orally.

In some embodiments, a compound of the present invention (eg, a factor XIa or kallikrein inhibitor) may be administered once or several times per day. The duration of treatment may be, for example, when administered once daily, a period of about 1, 2, 3, 4, 5, 6, 7 days or more. In some embodiments, the treatment is ongoing (eg, lifelong). In some embodiments, the administration is either a single dose as a single dosage form, multiple dosage forms of a smaller dose, or multiple administrations of divided doses at specified intervals. For example, the dosage form may be administered over about 0 hours to about 1 hour, about 1 hour to about 24 hours, about 1 to about 72 hours, about 1 to about 120 hours, or about 24 hours to at least about 120 hours. hours after injury. Alternatively, the dosage form may be administered over about 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 30, 40, 48, 72, 96, 120 hours after injury. Subsequent dosage forms may be administered at any time after the initial administration so that a therapeutic effect is achieved. In some embodiments, the initial dose is administered orally. In some embodiments, subsequent doses after the initial dose are administered parenterally (e.g., nasally, intramuscular-buccal, inhalation, implantation, percutaneous, intravenous (e.g., intravenous infusion, intravenous bolus injection), subcutaneous, intradermal, nasal, pulmonary, transdermal, intra-articular, intra-arterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or other infusion methods); orally; or rectally.

In some embodiments, the compounds of the present invention (eg, factor XIa or kallikrein inhibitor) are administered orally, eg, as a liquid or solid dosage form for ingestion over a period of about 5 minutes to about 1 week; from about 30 minutes to about 24 hours, from about 1 hour to about 12 hours, from about 2 hours to about 12 hours, from about 4 hours to about 12 hours, from about 6 hours to about 12 hours, from about 6 hours to approximately 10 hours; from about 5 minutes to about 1 hour, from about 5 minutes to about 30 minutes; from about 12 hours to about 1 week, from about 24 hours to about 1 week, from about 2 days to about 5 days, or from about 3 days to about 5 days. In one embodiment, a compound of the present invention (eg, a factor XIa or kallikrein inhibitor) is administered orally in a liquid dosage form. In another embodiment, a compound of the present invention (eg, a factor XIa or kallikrein inhibitor) is administered orally in a solid dosage form.

If a patient undergoing therapy experiences an incomplete response or relapse after completion of the first cycle of therapeutic treatment, subsequent courses of therapy may be required to achieve a limited or complete therapeutic response (e.g., chronic treatment, e.g., lifelong).

In some embodiments, a compound of the present invention (eg, a factor XIa or kallikrein inhibitor) is administered intravenously, eg, as an intravenous infusion or intravenous bolus injection, over a period of about 5 minutes to about 1 week; from about 30 minutes to about 24 hours, from about 1 hour to about 12 hours, from about 2 hours to about 12 hours, from about 4 hours to about 12 hours, from about 6 hours to about 12 hours, from about 6 hours to approximately 10 hours; from about 5 minutes to about 1 hour, from about 5 minutes to about 30 minutes; from about 12 hours to about 1 week, from about 24 hours to about 1 week, from about 2 days to about 5 days, or from about 3 days to about 5 days. In one embodiment, a compound of the present invention (eg, a factor XIa or kallikrein inhibitor) is administered as an intravenous infusion over about 5, 10, 15, 30, 45 or 60 minutes or longer; about 1, 2, 4, 6, 8, 10, 12, 16 or 24 hours or longer; approximately 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 days or longer.

In some embodiments, the compound is administered after an ischemic attack. In some embodiments, the compound is administered approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days or more after the ischemic attack. In some embodiments, the compound is administered approximately 1, 2, 3, 4, 5, 6, 7, or 8 weeks or more after the ischemic attack. In some embodiments, the compound is administered approximately 1, 2, 3, 4, 5, or 6 months or more after the ischemic attack.

In some embodiments, the compound is administered in combination with an additional therapeutic agent. In some embodiments, the additional therapeutic agent is administered after administration of the compound. In some embodiments, the additional therapeutic agent is administered orally. In some embodiments, the additional therapeutic agent is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 16, 18, 20, or 24 hours or more after administration of the compound. In some embodiments, the additional therapeutic agent is administered at least 1, 2, 3, 4, 5, 6, 7, 14, 21, or 28 days or more after administration of the compound. In some embodiments, the additional therapeutic agent is administered about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days or more after administration of the compound. In some embodiments, the additional therapeutic agent is administered continuously after administration of the compound. In some embodiments, the additional therapeutic agent is an NSAID, a platelet aggregation inhibitor, or an anticoagulant. In some embodiments, the additional therapeutic agent results in an additive therapeutic effect. In some embodiments, the additional therapeutic agent results in a synergistic therapeutic effect.

Doses and treatment regimens

The effective amount of the small molecule factor XIa inhibitor or kallikrein administered in accordance with the present invention can be determined by one skilled in the art. The specific dosage and frequency of administration for any given patient may vary and will depend on various factors, including the potency of the particular compound used, the metabolic stability and duration of action of that compound, the species, age, body weight, general health, sex and dietary intake of the patient, route and timing of administration, rate of elimination, combination medications, and severity of the specific condition.

As the patient's condition improves, a maintenance dose of a compound, composition, or combination provided herein may be administered if necessary. Subsequently, the dosage or frequency of administration, or both, can be reduced depending on the symptoms to a level at which the improved condition is maintained when the symptoms are reduced to the desired level. However, patients may require periodic treatment on a long-term basis for any recurrence of disease symptoms.

EXAMPLES

The starting materials and various intermediates described in the following examples can be obtained from commercial sources, from commercially available organic compounds, or using known synthetic methods. Typical examples of methods suitable for preparing intermediates of the invention are also given below.

General techniques

All non-aqueous reactions were carried out under nitrogen atmosphere to maintain an anhydrous atmosphere and obtain maximum yields. All reaction mixtures were stirred using a top drive stirrer or a Teflon coated magnetic stirrer. The term "dried over" refers to drying a solution of the reaction product over a specific drying agent, such as magnesium sulfate or sodium sulfate, and then filtering the solution through a suitable filter paper or Schott funnel. The terms "concentrate", "concentrate under reduced pressure" or "evaporate" refer to the removal of solvents under reduced pressure using a rotary evaporator. The terms "chromatography" or "chromatography" refer to the use of silica gel flash column chromatography unless otherwise noted. Flash chromatography can be performed using gas pressure (such as nitrogen) or using a mechanical solvent pump under pressure, such as a commercial system from Biotage or other suppliers. The term "preparative TLC" refers to thin layer chromatography on silica gel plates. The term "flash chromatography" refers to pressurized column chromatography and can be performed using commercial units such as the one from Biotage. The term "retention time" refers to the elution time of a product by HPLC under specific conditions. Unless otherwise noted, all NMR spectra are proton ( ^1H ) NMR spectra in the indicated solvent.

In the following examples, high performance liquid chromatography/mass spectrometry (HPLC/MS) to characterize compounds is performed under the following conditions:

1. HPLC/MS Equipment: Analysis is performed using a Waters 2545 Binary Gradient Module (Waters Corporation, Milford, MA), Waters SFO System Fluidics Organizer, Waters 2996 Diode Array Detector and Waters 2767 Autosampler, 3100 Mass Detector. Data is acquired using MassLynxTM 4.0 software with processing using OpenLynxTM and AutoLynxTM.

2. Analytical HPLC conditions: 4.6 × 50 mm column; UV 10 spectra/sec., 220-340 nm; volumetric flow rate 2.0 ml/min; injection volume 5 µl;

gradient conditions A: mobile phase A - water with 0.1% formic acid; mobile phase B - acetonitrile with 0.1% formic acid; gradient: 1.50 min. from 99.0% A to 95.0% B; 0.5 min. holding; then return to 99.0% A within 0.5 minutes;

gradient conditions B: mobile phase A - water with 0.1% formic acid; mobile phase B - acetonitrile with 0.1% formic acid, gradient: 3.00 min. from 95.0% A to 95.0% B; 2.0 min. holding; then return to 95.0% A within 0.5 min.

The meanings of the abbreviations used in the experimental examples are presented in the table below.

Table of abbreviations

ACN Acetonitrile Ammonium chloride solution Saturated aqueous ammonium chloride solution Aqueous ammonium chloride solution Saturated aqueous ammonium chloride solution Aqueous NaHCO ₃ solution Saturated aqueous sodium bicarbonate solution Saturated salt solution Saturated aqueous sodium chloride solution VOS or Vos Tert-butoxycarbonyl Celite® Diatomaceous earth DBU 1,8-Diazabicyclo[5.6.0]undec-7-ene DXM Dichloromethane DMSO Dimethyl sulfoxide EA Ethyl acetate Ether Diethyl ether Hour. Watch HATU 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate LC-MS HPLC/MS LDA Lithium diisopropylamide Min. minutes NMR Nuclear magnetic resonance Pd/C Palladium on coal RMV 4-Methoxybenzyl CT Room temperature TBME tert-Butyl methyl ether TEA Triethylamine Tret Tertiary THF Tetrahydrofuran TMAF Tetramethylammonium fluoride ACN Acetonitrile DMF N,N-Dimethylformamide TLC Thin layer chromatography X-phos 2-Dicyclohexylphosphino-2’,4’,6’-triisopropylbiphenyl Chromatography or Chromatography Purification of products using silica gel flash chromatography Concentrate or concentrate in vacuo Concentration of organic solutions under reduced pressure using a rotary evaporator

Scheme 1

Scheme 1 illustrates a general method for the preparation of (2S,3R)-trans-disubstituted-4-oxoazetidines of general structure 10 containing an amide group at the 2-position. Alkylation of the trans dianion 1 with commercially available or readily synthesized compound 2 (as described in the preparation of intermediates 7 and 8) results in the desired compound 3 as a predominantly trans product while retaining the stereochemistry at the 2 position (Baldwin, J.E., et al. , Tetrahedron, 1990, 46, 4733). Removal of the TBS protecting group at N-1 results in compound 4, which is then reacted with isocyanate 5 or phenylcarbamate 6 to yield compound 7. Coupling of carboxylic acid 7 with a commercially available or easily synthesized amine 8 (as described in the preparation of intermediate 6) in the presence of a base and a coupling reagent results in compound 9. Subsequent removal of the protecting groups P1 and P2 results in compound 10.

Scheme 2

Scheme 2 illustrates an alternative method for preparing compound 10. Coupling of carboxylic acid 3 with a commercially available or readily synthesized amine 8 (as described in the preparation of intermediate 6) in the presence of a base and coupling reagent results in compound 11. Removal of the TBS protecting group at N-1 yields compound 12, which is then reacted with isocyanate 5 or phenylcarbamate 6 yields compound 9. Subsequent removal of the protecting groups P1 and P2 provides an example of the general compound 10.

Scheme 3

Scheme 3 illustrates the method for preparing compound 18. Alkylation of the dianion of compound 1 with commercially available or easily synthesized compound 13 (prepared as described for the preparation of intermediate 9) leads to the desired compound 14. Coupling of carboxylic acid 14 with commercially available or easily synthesized amine 8 ( as described in Preparation of Intermediate 6) in the presence of a base and coupling reagent yields compound 15. Removal of the TBS protecting group at N-1 yields compound 16, which is then reacted with isocyanate 5 or phenylcarbamate 6 to yield compound 17. Subsequent Removal of the P2 protecting group results in compound 18.

Scheme 4

Scheme 4 illustrates the method for preparing compound 19. Alkylation of compound 18 (R ₅ =hydrogen or alkyl) using an alkyl halide in the presence of a base or the reaction of compound 18 with an alkyl aldehyde in the presence of a reducing agent (such as NaBH ₃ CN, etc.) produces connections 19.

Scheme 5

Scheme 5 illustrates the method for preparing compound 21. Alkylation of compound 9 using an alkyl halide in the presence of a base gives compound 20. Subsequent removal of the protecting groups P1 and P2 leads to compound 21.

Example 1. Preparation of intermediate compounds.

Intermediate 1: Phenyl-N-[(1S)-1-cyclohexyl-2,2,2-trifluoroethyl]carbamate

To a stirred solution of phenylchloroformate (714 μL, 5.69 mmol) in dry THF (30 mL) at 0°C, add a solution of (1S)-1-cyclohexyl-2,2,2-trifluoroethanamine(1, 00 g, 5.52 mmol) and pyridine (536 µl, 6.63 mmol) in THF (18 ml) After addition for 45 minutes at 0°C, the mixture is diluted with ethyl acetate (EA) (50 ml), washed with aqueous NaHCO solution ₃ (50 ml), water (40 ml) and brine (30 ml), dry over MgSO ₄ and concentrate. The resulting solid was triturated with hexanes (35 ml), collected by filtration, washed with small amounts of hexanes and dried to give 1.51 g (91%) of the title compound as a white solid, which was used without further purification. ^1H NMR (400 MHz, CDCl ₃ ) δ 7.40 (m, 2H), 7.24 (m, 1H), 7.17 (m, 2H), 5.20 (br. d, 1H), 4 .25 (m, 1H), 1.92-1.68 (m, 6H), 1.40-1.05 (m, 5H); MS (ESI+) m/z 302.1 (M+H) ⁺ , retention time: 2.03 min. (method A).

Intermediate 2: Phenyl N-[(1R)-1-cyclohexylpropyl]carbamate

Scheme 6

Step 1. Preparation of [N(E),S(S)]-N-(cyclohexylmethylene)-2-methyl-2-propanesulfonamide

Titanium (IV) ethoxide (101, 7 g, 0.446 mol). The resulting reaction mixture was heated and maintained at 75°C for 2 hours. The mixture is cooled to RT. Add brine (250 ml) and the mixture is stirred vigorously, then filtered through Celite® and washed with EA (300 ml). The aqueous layer is extracted with EA (2 x 200 ml). The combined organic phase is dried over MgSO ₄ , filtered and concentrated. The crude product was purified by chromatography (elution: 4:1 hexane:EA) to give the product as a colorless oil (45.6 g, 95%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.97 (1H, d, J=4.6 Hz), 2.47 (1H, m), 1.92-1.68 (7H, m), 1.67 -1.35 (3H, m), 1.20 (9H, s).

Step 2. Preparation of [S(S)]-N-((1R)-1-cyclohexylpropyl)-2-methyl-2-propanesulfonamide

[N(E),S(S)]-N-(cyclohexylmethylene)-2-methyl-2-propanesulfonamide (32.3 g, 0.15 mol) was dissolved in TBME (500 ml). The reaction mixture was cooled to -40°C, ethylmagnesium bromide (3M in ether, 100 ml, 0.3 mol) was added dropwise to the mixture and the reaction mixture was stirred at -25°C for 4 hours. Add aqueous ammonium chloride and then EA. The layers are separated and the aqueous layer is extracted with EA twice. The combined organic phase was washed with brine, dried over sodium sulfate, concentrated and purified by chromatography (elution: 2:1 hexane:EA) to give [S(S)]-N-((1R)-1-cyclohexylpropyl)-2 -methyl-2-propanesulfonamide as a colorless oil (22 g, 60%). ^1H NMR (400 MHz, CDCl ₃ ) δ 0.90-1.00 (m, 4H), 1.12 (m, 1H), 1.20 (s, 9H), 1.44 (m, 1H ), 1.52-1.80 (m, 9H), 2.88-3.00 (m, 2H).

Step 3. Preparation of (R)-1-cyclohexylpropan-1-amine hydrochloride

[S(S)]-N-((1R)-1-cyclohexylpropyl)-2-methyl-2-propanesulfonamide (31.86 g, 0.129 mol) was dissolved in methanol (200 ml) and 4 M HCl in 1 was added slowly. 4-dioxane (100 ml). The reaction solution is stirred for 60 minutes. The reaction mixture is concentrated to obtain a solid residue. Hexane (200 ml) was added to the solid residue and the resulting suspension was stirred at RT for 60 minutes. The solid was collected by filtration, washed with hexanes and dried to give (R)-1-cyclohexylpropan-1-amine hydrochloride as a white solid (12.8 g, 56%).

Step 4. Preparation of phenyl-N-[(1R)-1-cyclohexylpropyl]carbamate

To a stirred suspension of (R)-1-cyclohexylpropan-1-amine hydrochloride (5 g, 28.1 mmol) in DCM (100 ml) at 0°C add TEA (6.82 g, 67.4 mmol) and then slowly add phenyl chloroformate (4.84 g, 31 mmol). After stirring for an additional 45 min. At RT, the mixture is diluted with DCM (50 ml), washed with aqueous NaHCO ₃ (50 ml), water (40 ml) and brine (30 ml), dried over MgSO ₄ and concentrated. The resulting solid residue is triturated with hexane (35 ml). The solid was collected by filtration, washed with small amounts of hexane and dried to give 6.3 g (85%) of the title compound as a white solid, which was used without further purification. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.40 (m, 2H), 7.24 (m, 1H), 7.17 (m, 2H), 4.75 (br. d, 1H), 3 .50 (m, 1H), 1.92-1.68 (m, 6H), 1.50-1.05 (m, 10H). MS (ESI+) 262.2 (M+H) ⁺ , retention time: 2.09 min. (method A).

Intermediate 3. (R)-1-(3-pyridinyl)-1-propanamine dihydrochloride

Scheme 7

Step 1. Preparation of [S(S)]-N-[1-(pyridin-2-yl)propylidene]-2-methylpropane-2-sulfinamide

A solution of 3-propionylpyridine (5 g, 37 mmol), (S)-(-)-2-methylpropane-2-sulfinamide (4.7 g, 38.8 mmol) and titanium (IV) ethoxide (17.8 g, 78 mmol) in THF (100 ml) is heated and maintained at 80°C for 3 days. After cooling, add brine (100 ml) and EA (100 ml). The mixture is stirred at RT for 1 hour. The mixture is filtered through Celite® and the filter is washed with EA. The organic phase is separated, concentrated and the residue is purified by chromatography (elution: 2:1 hexane:EA) to give the product as a colorless oil (8.15 g, 92%).

Step 2. Preparation of [S(S),R]-N-[1-(pyridin-2-yl)propyl]-2-methylpropane-2-sulfinamide

To a solution of [(S(S)]-N-[1-(pyridin-2-yl)propylidene]-2-methylpropane-2-sulfinamide (8.15 g, 34.2 mmol) in THF (150 ml) at -78°C add a solution of tri-sec-butylborohydride (L-selectride, 1M solution in THF, 68 ml, 68 mmol). The reaction mixture is stirred for 3 hours at -78°C. The reaction mixture is quenched with aqueous ammonium chloride (150). ml) The mixture is filtered through Celite® and washed with EA. The organic phase is separated, dried over Na ₂ SO ₄ and the residue is purified by chromatography (elution: 1:1 hexane:EA) to give the product as a colorless oil (4.3). g, 53%).

Step 3. (R)-1-(3-pyridinyl)-1-propanamine dihydrochloride

To a solution of [S(S),R]-N-[1-(pyridin-2-yl)propyl]-2-methylpropan-2-sulfinamide (4.3 g, 17.9 mmol) in methanol (50 ml) slowly add 4M HCl in 1,4-dioxane (22 ml). The reaction mixture is stirred for 1 hour. The reaction mixture is concentrated to obtain a solid residue. Hexane (50 ml) was added to the solid residue and the resulting suspension was stirred at RT for 1 hour. The solid was collected by filtration, washed with hexanes and dried to give (R)-1-(3-pyridinyl)-1-propanamine dihydrochloride as a white solid (3.3 g, 90%). ^1H NMR (400 Hz, DMSO- _d6 ) δ 0.80 (t, 2H), 1.90 (m, 1H), 2.10 (m, 1H), 4.20 (m, 1H), 8 .04 (d, 1H), 8.80 (d, 1H), 8.90 (m, 2H), 9.20 (br., 4 H), MS (ESI+) m/z 137.1 (M+ H) ⁺ . Retention time: 0.35 min. (method A).

Intermediate 4. (S)-1-Cyclopropyl-2,2,2-trifluoroethylamine hydrochloride

Scheme 8

Step 1. Preparation of [N(E),R(S)]-N-(cyclopropylmethylene)-2-methyl-2-propanesulfonamide

To a solution of cyclopropanecarboxaldehyde (10 g, 0.14 mol) in THF (100 ml) add (R)-2-methyl-2-propanesulfonamide (18.2 g, 0.15 mol), and titanium (IV) ethoxide (65 .1 g, 0.285 mol). The resulting mixture is heated and kept at 75°C for 2 hours. The mixture is cooled to RT. Add brine (150 ml) and stir vigorously; the mixture is then filtered through Celite® and the filter bed is washed with EA (300 ml). The aqueous layer is extracted with EA (2 x 200 ml). The combined organic extracts are dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 4:1 hexane:EA) to give the product as a colorless oil (22.2 g, 90%).

Step 2. Preparation of [R(S)]-N-((1S)-1-cyclopropyl-2,2,2-trifluoroethyl)-2-methyl-2-propanesulfonamide

To a solution of [N(E),R(S)]-N-(cyclopropylmethylene)-2-methyl-2-propanesulfonamide (7.8 g, 45 mmol) in THF (150 ml) add TMAF (5 g, 53, 7 mmol). The solution is degassed with nitrogen and then cooled to -55°C. A solution of trifluoromethyltrimethylsilane (9.6 g, 67.5 mmol) in THF (150 ml) was added dropwise and the reaction mixture was stirred at -55°C for 2 hours. The reaction mixture is allowed to slowly warm to -10°C and then the reaction is quenched with an aqueous solution of ammonium chloride. The aqueous layer was extracted with EA and the combined organic layer was dried and concentrated to give a yellow oil which was used without further purification (10.7 g, 90%).

Step 3. Preparation of (S)-1-cyclopropyl-2,2,2-trifluoroethylamine hydrochloride

[R(S)]-N-((1S)-1-cyclopropyl-2,2,2-trifluoroethyl)-2-methyl-2-propanesulfonamide (10.7 g, 44 mmol) dissolved in methanol (100 ml) and slowly add 4M HCl in 1,4-dioxane (44 ml). The solution is stirred for 1 hour. The solvent is evaporated to obtain a solid residue. Hexane (100 ml) was added to the solid residue to obtain a suspension, which was stirred at RT for 1 hour. The solid was collected by filtration, washed with hexanes and dried to give (S)-1-cyclopropyl-2,2,2-trifluoroethylamine hydrochloride as a white solid (5.4 g, 70%). ¹ H NMR (400 Hz, DMSO-d6) δ 0.50-0.70 (m, 4H), 1.04 (m, 1H), 3.60 (m, 1H) 9.20 (br., 3H ).

Intermediate 5. Synthesis of (R)-(+)-1-phenylpropyl isocyanate

To a solution of R-(+)-1-phenylpropylamine (5 g, 37 mmol) in DCM (80 ml) was added 1N aqueous NaHCO ₃ (86 ml). The mixture is cooled to 0°C and triphosgene (3.73 g, 12.6 mmol) is added. The reaction mixture is stirred for 15 minutes. at 0°C. The reaction mixture is extracted twice with DCM. The combined organic phase was dried over Na ₂ SO ₄ and concentrated to give (R)-(+)-1-phenylpropyl isocyanate as a colorless oil (5.4 g, 91%), which was used in the next step without further purification. ^1H NMR (400 Hz, CDCl ₃ ) δ 0.97 (t, 3H), 1.85 (m, 2H), 4.52 (t, 1H), 7.29 (m, 3H), 7.34 (m, 2H).

Intermediate 6: N,1-Dimethyl-1H-pyrazol-5-amine hydrochloride

Scheme 9

Step 1. Preparation of tert-butyl-N-(1-methyl-1H-pyrazol-5-yl)carbamate

To a solution of 1-methyl-1H-pyrazol-5-ylamine (5 g, 51 mmol) in DCM (30 mL) at RT, add di-tert-butyl dicarbonate (12.4 g, 56.8 mmol) followed by TEA (10 .4 g, 103 mmol). The resulting mixture was stirred at RT overnight. The mixture is washed with water. The organic layer was dried and evaporated to give the product as a white solid (10 g, 98%), which was used in the next step without further purification. ^1H NMR (400 Hz, CDCl ₃ ) δ 1.40 (s, 9H), 1.74 (s, 1H), 3.16 (s, 3H), 6.02 (d, 1H), 7.40 (d, 1H).

Step 2. Preparation of tert-butyl-N-methyl-N-(1-methyl-1H-pyrazol-5-yl)carbamate

To a solution of tert-butyl-N-(1-methyl-1H-pyrazol-5-yl)carbamate (10 g, 51 mmol) in THF (100 ml) at 0°C add sodium hydride (2.5 g, 60% in mineral oil). The mixture is allowed to warm to RT and the mixture is stirred for 30 minutes. Iodomethane (10.8 g, 76 mmol) was slowly added to the reaction mixture. The resulting mixture was stirred at RT overnight. The reaction mixture is quenched with an aqueous solution of ammonium chloride (30 ml). The mixture is extracted with EA (50 ml x 3). The combined organic phases are washed with brine, dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 4:1 hexane:EA) to give the product as a colorless oil (9.5 g, 89%). ^1H NMR (400 Hz, CDCl ₃ ) δ 1.40 (s, 9H), 3.16 (s, 3H), 3.64 (s, 3H), 6.10 (d, 1H), 7.42 (d, 1H).

Step 2. Preparation of N,1-dimethyl-1H-pyrazod-5-amine hydrochloride

To a solution of tert-butyl-N-methyl-N-(1-methyl-1H-pyrazol-5-yl)carbamate (9.5 g, 45 mmol) in DCM (50 ml) add 4N HCl in dioxane (45 ml) . The resulting mixture was stirred at RT overnight. Solvents were removed in vacuo to give the product as a white solid (6 g, 90%). ^1H NMR (400 Hz, DMSO-d6) δ 2.80 (s, 3H), 3.64 (s, 3H), 4.20 (br., 2H), 5.84 (d, 1H), 7 .94 (d, 1H).

Intermediate 7: tert-Butyl-[4-(bromomethyl)-6-methylpyridin-2-yl](4-methoxybenzyl)carbamate

Scheme 10

Step 1. Preparation of methyl 2-((tert-butoxycarbonyl)amino)-6-methylpyridine-4-carboxylate

A mixture of methyl 2-chloro-6-methylpyridine-4-carboxylate (20 g, 107.8 mmol), tert-butyl carbamate (15.2 g, 129.7 mmol), cesium carbonate (70.2 g, 215,. 5 mmol), X-phos (5.14 g, 10.8 mmol) and palladium acetate (1.2 g, 5.3 mmol) in 1,4-dioxane (300 ml) are degassed with nitrogen and then stirred at 90° C in a nitrogen atmosphere for two hours. The mixture is cooled to RT, diluted with water (300 ml) and then extracted with EA (150 ml x 2). The organic extracts are combined and washed with brine, dried (MgSO ₄ ) and concentrated. The residue was purified by chromatography (elution: 0-30% EA in hexane) to give methyl 2-((tert-butoxycarbonyl)amino)-6-methylpyridine-4-carboxylate as a white solid (25.8 g, 90%) . ^1H NMR (400 Hz, CDCl ₃ ) δ 1.52 (s, 9H), 2.48 (s, 3H), 3.92 (s, 3H), 7.36 (s, 1H), 7.40 (s, 1H), 8.24 (s, 1H).

Step 2. Preparation of methyl 2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridine-4-carboxylate

To a solution of methyl 2-((tert-butoxycarbonyl)amino)-6-methylpyridine-4-carboxylate (12.3 g, 46.2 mmol) in DMF (100 ml) add potassium tert-butoxide (7.3) in one portion g, 60.2 mmol). The mixture is stirred at RT for 5 minutes. and then p-methoxybenzyl chloride (9.8 g, 59.9 mmol) was added. The resulting mixture was stirred at RT for 12 hours. Add water (150 ml) and extract the resulting mixture with EA (100 ml x 2). The combined organic phase is washed with brine, dried (MgSO ₄ ) and concentrated. The residue was purified by chromatography (elution: 0-20% EA in hexane) to give methyl 2-((t-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridine-4-carboxylate as a white solid (12.8 g, 72%). MS (ESI ⁺ ) m/z 387.2 (M+H) ⁺ , retention time: 2.15 min. (method A) . ^1H NMR (400 Hz, CDCl ₃ ) δ 1.45 (s, 9H), 2.52 (s, 3H), 3.76 (s, 3H), 3.91 (s, 3H), 5.14 (s, 2H), 6.78 (d, 2H), 7.22 (d, 2H), 7.38 (s, 1H), 7.97 (s, 1H).

Step 3. Preparation of tert-butyl-(4-(hydroxymethyl)-6-methylpyridin-2-yl)(4-methoxybenzyl)carbamate

To a solution of methyl 2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridine-4-carboxylate (12.8 g, 33.1 mmol) in THF (100 ml) add lithium borohydride (1 .5 g, 68.9 mmol). The mixture is heated and kept at 40°C for 4 hours. The reaction mixture is cooled to 0°C and carefully quenched by slowly adding dropwise an aqueous solution of NaHCO ₃ (30 ml). The mixture is diluted with water (100 ml) and extracted with EA (50 ml x 2). The combined organic phase is washed with brine (50 ml), dried (MgSO ₄ ) and concentrated. The residue was purified by chromatography (elution: 40% EA in hexanes) to give tert-butyl (4-(hydroxymethyl)-6-methylpyridin-2-yl)(4-methoxybenzyl)carbamate as a colorless oil (11.3 g, 95 %). MS (ESI ⁺ ) m/z 359.1 (M+H) ⁺ , retention time: 1.76 min. (method A). ^1H NMR (400 Hz, CDCl ₃ ) δ 1.40 (s, 9H), 2.48 (s, 3H), 3.76 (s, 3H), 4.62 (m, 2H), 5.14 (s, 2H), 6.80 (d, 2H), 6.84 (s, 1H), 7.20 (d, 2H), 7.40 (s, 1H).

Step 4. Preparation of tert-butyl-(4-(bromomethyl)-6-methylpyridin-2-yl)(4-methoxybenzyl)carbamate

To a solution of tert-butyl-(4-(hydroxymethyl)-6-methylpyridin-2-yl)(4-methoxybenzyl)carbamate (11.3 g, 31.5 mmol) and carbon tetrabromide (12.3 g, 37.1 mmol) in DCM (150 ml) at 0°C add triphenylphosphine (9.7 g, 37.1 mmol) in one portion. The reaction mixture is stirred for 20 minutes, the mixture is allowed to warm to RT and then the mixture is stirred for one hour. The mixture is diluted with DCM (100 ml) and extracted with aqueous NaHCO ₃ solution (50 ml x 1). The organic layer is dried (MgSO ₄ ) and concentrated. The residue was purified by chromatography (elution: 0-20% EA in hexane) to give tert-butyl (4-(bromomethyl)-6-methylpyridin-2-yl)(4-methoxybenzyl)carbamate as a white solid (10.6 g, 80%). MS (ESI+) m/z 422.1 (M+H) ⁺ . Retention time: 2.18 min. (method A). ^1H NMR (400 Hz, CDCl ₃ ) δ 1.40 (s, 9H), 2.46 (s, 3H), 3.78 (s, 3H), 4.38 (s, 2H), 5.14 (s, 2H), 6.80 (d, 2H), 6.84 (s, 1H), 7.22 (d, 2H), 7.46 (s, 1H).

Intermediate 8: 4-(bromomethyl)-N,N-bis(4-methoxybenzyl)pyridin-2-amine

Scheme 11

Step 1. Preparation of 2-[bis-(4-methoxybenzyl)amino]isonicotinic acid methyl ester

A mixture of 2-aminopyridine-4-carboxylic acid methyl ester (5.5 g) and 4-methoxybenzyl chloride (14.64 g) in 33 ml of acetonitrile is refluxed for 3 hours; TEA (7.3 g) is then slowly added to the boiling mixture. The reaction mixture is allowed to cool to room temperature and the mixture is stirred overnight. The reaction mixture was concentrated by removing the solvents, and water was added to the residue to form a solid precipitate. The solids were collected by filtration and recrystallized from isopropanol to give 2-[bis-(4-methoxybenzyl)amino]isonicotinic acid methyl ester (3 g, 21%) as a white solid.

Step 2. Preparation of {2-[bis-(4-methoxybenzyl)amino]pyridin-4-yl}methanol

Lithium aluminum hydride (388 mg) is added in portions to THF (16 ml), pre-cooled to 0°C. 2-[bis(4-methoxybenzyl)amino]isonicotinic acid (4 g) in THF (16 ml) was then added dropwise to the mixture, maintaining the temperature of the mixture between 0 and -5°C. After this, EA (900 mg), water (388 mg) and 15% NaOH aqueous solution (388 mg) were slowly added to the reaction mixture successively. The mixture is stirred for 10 minutes. and then add anhydrous Na ₂ SO ₄ (1.3 g). The mixture is stirred for 30 minutes. The solid precipitate was collected by filtration and washed with THF (12 ml). The filtrate is concentrated to obtain {2-[bis(4-methoxybenzyl)amino]pyridin-4-yl}methanol, which is used in the next step without further purification

Step 3. Preparation of 4-(bromomethylpyridin-2-yl)-bis(4-methoxybenzyl)amine

A solution of {2-[bis(4-methoxybenzyl)amino]pyridin-4-yl}methanol (5 g) and CBr ₄ (5 g) in DCM (25 ml) is cooled to 0-10°C. A solution of PPh ₃ (4.32 g) in DCM (10 ml) was added dropwise. The reaction mixture is analyzed by TLC. If the starting alcohol is present, add additional PPh ₃ until the reaction is complete. The reaction mixture was concentrated, the oily residue was treated with 50% aqueous ethanol (16 ml) and stirred for 1 hour at RT. The solid is filtered and washed with a small amount of 50% aqueous ethanol. The resulting filter cake was suspended with stirring in 50% aqueous ethanol (8 ml) for 1 hour at RT. The solid was again filtered and dried to obtain the title compound as an off-white crystalline solid (5.2 g). ^1H NMR ( _CDCl3 ): 8.17 (d, 1H), 7.14 (d, 4H), 6.84 (d, 4H), 6.60 (d, 1H), 6.45 (s, 1H), 4.69 (s, 4H), 4.23 (s, 2H), 3.79 (s, 6H). MS (ESI+) m/z 427.2 (M+H) ⁺ ; retention time: 1.2 min. (method A).

Intermediate 9. tert-Butyl-(4-(bromomethyl)pyridin-2-yl)methylcarbamate

Scheme 12

Step 1. Preparation of methyl 2-(tert-butoxycarbonyl)(methyl)amino)isonicotinate

A suspension of methyl 2-((tert-butoxycarbonyl)amino)isonicotinate (50.4 g, 200 mmol) in DMF (500 ml) is cooled to 0°C and sodium hydride (10.4 g, 60% in mineral oil) is added in portions , 260 mmol). The mixture is allowed to warm to RT and the mixture is stirred for 30 minutes. Iodomethane (37.2 g, 262 mmol) was slowly added to the mixture. The reaction mixture was stirred at RT overnight. The reaction mixture is quenched by adding aqueous ammonium chloride (100 ml) and diluted with water (400 ml). The mixture is extracted with EA (250 ml x 2). The combined organic phases are washed with water (100 ml) and brine (100 ml), dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 5:1 hexane:EA) to give the product as a colorless oil (48.9 g, 92%). ^1H NMR (400 Hz, CDCl ₃ ) δ 1.54 (s, 9H), 3.42 (s, 3H), 3.94 (s, 3H), 7.52 (d, 1H), 8.27 (s, 1H), 8.48 (d, 1H).

Step 2. Preparation of tert-butyl-(4-(hydroxymethyl)pyridin-2-yl)methylcarbamate

To a solution of methyl 2-(tert-butoxycarbonyl)(methyl)amino)isonicotinate (48.48 g, 182.1 mmol) in THF (500 ml) add lithium borohydride (5.15 g, 236.5 mmol) at RT. . The mixture is heated and kept at 40°C for 4 hours. The reaction mixture is cooled to 0°C and carefully quenched by slowly adding dropwise aqueous NaHCO ₃ solution (50 ml). The mixture is diluted with water (500 ml) and extracted with EA (250 ml x 2). The combined organic phase is washed with brine (150 ml), dried (MgSO ₄ ) and concentrated. The residue was purified by chromatography (elution: 40% EA in hexane) to give tert-butyl (4-(hydroxymethyl)pyridin-2-yl)methylcarbamate as a colorless oil (41.2 g, 95%). MS (ESI+) m/z 239.1 (M+H) ⁺ , retention time: 1.36 min. (method A). ^1H NMR (400 Hz, CDCl ₃ ) δ 1.54 (s, 9H), 3.40 (s, 3H), 4.68 (s, 2H), 7.02 (d, 1H), 7.64 (s, 1H), 8.34 (d, 1H).

Step 3. Preparation of tert-butyl-(4-(bromomethyl)pyridin-2-yl)methylcarbamate

To a solution of tert-butyl-(4-(hydroxymethyl)pyridin-2-yl)methylcarbamate (20.52 g, 86.1 mmol) and carbon tetrabromide (33.41 g, 100.7 mmol) in DCM (400 ml) At 0°C, add triphenylphosphine (26.43 g, 100.7 mmol) in one portion. The reaction mixture is stirred for 20 minutes, the mixture is allowed to warm to RT and then stirred for one hour. The mixture was diluted with DCM (100 ml) and washed with aqueous NaHCO ₃ (150 ml) and brine (150 ml). The organic layer is dried (MgSO ₄ ) and concentrated. The residue was purified by chromatography (elution: 0-20% EA in hexane) to give tert-butyl (4-(bromomethyl)pyridin-2-yl)methylcarbamate as a colorless oil (20.7 g, 80%). ^1H NMR (400 MHz, CDCl ₃ ) δ 1.54 (s, 9H), 3.41 (s, 3H), 4.39 (s, 2H), 6.92-7.12 (m, 1H) , 7.66-7.84 (m, 1H), 8.24-8.43 9 (m, 1H).

Intermediate 10. tert-Butyl-(4-(bromomethyl)-6-methylpyridin-2-yl)methylcarbamate

Scheme 13

Step 1. Preparation of methyl 2-(tert-butoxycarbonyl)(methyl)amino)-6-methylpyridine-4-carboxylate

A suspension of methyl 2-((tert-butoxycarbonyl)amino)-6-methylpyridine-4-carboxylate (20 g, 75.2 mmol) in DMF (200 ml) is cooled to 0°C and sodium hydride (3, 9 g, 60% in mineral oil, 97.5 mmol). The mixture is allowed to warm to RT and the mixture is stirred for 30 minutes. Iodomethane (12.8 g, 90.1 mmol) was slowly added to the reaction mixture. The reaction mixture was stirred at RT overnight. The reaction is quenched by adding aqueous ammonium chloride (50 ml) and diluted with water (200 ml). The mixture is extracted with EA (250 ml x 2). The combined organic phases are washed with water (100 ml) and brine (100 ml), dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 5:1 hexane:EA) to give the product as a colorless oil (18.9 g, 90%). MS (ESI+) m/z 281.1 (M+H) ⁺ . Retention time: 2.03 min. (method A).

Step 2. Preparation of tert-butyl-(4-(hydroxymethyl)-6-methylpyridin-2-yl)methylcarbamate

To a solution of methyl 2-(tert-butoxycarbonyl)(methyl)amino)-6-methylpyridine-4-carboxylate (27 g, 96.4 mmol) in THF (300 ml) add lithium borohydride (2.1 g, 96.4 mmol). The mixture is heated and kept at 40°C for 4 hours. The reaction mixture is cooled to 0°C and carefully quenched by slowly adding dropwise aqueous NaHCO ₃ solution (50 ml). The mixture is diluted with water (200 ml) and extracted with EA (250 ml x 2). The combined organic phase is washed with brine (150 ml), dried (MgSO ₄ ) and concentrated. The residue was purified by chromatography (elution: 40% EA in hexane) to give tert-butyl (4-(hydroxymethyl)-6-methylpyridin-2-yl)methylcarbamate as a colorless oil (20 g, 82%). MS (ESI+) m/z 253.1 (M+H) ⁺ , retention time: 1.31 min. (method A).

Step 3. Preparation of tert-butyl-(4-(bromomethyl)-6-methylpyridin-2-yl)methylcarbamate

To a solution of tert-butyl-4-(hydroxymethyl)-6-methylpyridin-2-yl)methylcarbamate (20 g, 79.4 mmol) and carbon tetrabromide (30.8 g, 92.8 mmol) in DCM (400 ml) At 0°C, add triphenylphosphine (224.4 g, 93.0 mmol) in one portion. The reaction mixture is stirred for 20 minutes, the mixture is allowed to warm to RT and then stirred for one hour. The mixture was diluted with DCM (100 ml) and extracted with aqueous NaHCO ₃ (150 ml) and brine (150 ml). The organic layer is dried (MgSO ₄ ) and concentrated. The residue was purified by chromatography (elution: 0-20% EA in hexane) to give tert-butyl (4-(bromomethyl)-6-methylpyridin-2-yl)methylcarbamate as a colorless oil (20.7 g, 80%). MS (ESI+) m/z 315.2 (M+H) ⁺ , retention time: 2.02 min. (method A).

Intermediate 11: tert-Butyl-[4-(bromomethyl)-5-fluoropyridin-2-yl](4-methoxybenzyl)carbamate

Scheme 14

Step 1. Preparation of methyl 2-((tert-butoxycarbonyl)amino)-5-fluoropyridine-4-carboxylate

Mixture of methyl 2-bromo-5-fluoropyridine-4-carboxylate (6.64 g, 28.4 mmol), tert-butyl carbamate (4.0 g, 34.1 mmol), cesium carbonate (13.0 g, 39 .8 mmol), X-phos (657 mg, 1.1 mmol) and Pd(dba) ₂ (520 mg, 0.9 mmol) in 1,4-dioxane (100 ml) are purged with nitrogen and then stirred at 100° C in a nitrogen atmosphere for 6 hours. The mixture is cooled to RT, diluted with water (100 ml) and extracted with EA (150 ml x 2). The combined organic phase is washed with brine, dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 0-30% EA in hexane) to give methyl 2-((tert-butoxycarbonyl)amino)-5-fluoropyridine-4-carboxylate as a white solid (4.5 g, 56%) . MS (ESI+) m/z 271.1 (M+H) ⁺ , retention time: 1.81 min. (method A). ^1H NMR (400 MHz, CDCl ₃ ) δ 1.52 (s, 9H), 3.96 (s, 3H), 8.24 (s, 1H), 8.40 (d, 1H), 9.00 (s, 1H).

Step 2. Preparation of methyl 2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-5-fluoropyridine-4-carboxylate

To a solution of methyl 2-((tert-butoxycarbonyl)amino)-5-fluoropyridine-4-carboxylate (4.5 g, 16.6 mmol) in DMF (50 ml) add potassium tert-butoxide (2.33 g) in portions , 19.2 mmol). The mixture is stirred at RT for 5 minutes; p-methoxybenzyl chloride (3.2 g, 20.4 mmol) was then added. The resulting mixture was stirred at RT for 12 hours. Add water (50 ml) and extract with EA mixture (50 ml x 2). The combined organic phase is washed with brine, dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 0-20% EA in hexane) to give methyl 2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-5-fluoropyridine-4-carboxylate as a white solid (4.8 g, 75%). MS (ESI ⁺ ) m/z 391.0 (M+H) ⁺ , retention time: 2.09 min. (method A).

Step 3. Preparation of tert-butyl-(4-(hydroxymethyl)-5-fluoropyridin-2-yl)(4-methoxybenzyl)carbamate

To a solution of methyl 2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-5-fluoropyridine-4-carboxylate (7.7 g, 19.7 mmol) in THF (100 ml) add lithium borohydride ( 0.5 g, 23.0 mmol). The mixture is heated and kept at 40°C for 4 hours. The reaction mixture is cooled to 0°C and carefully quenched by slowly adding dropwise an aqueous solution of NaHCO ₃ (30 ml). The mixture is diluted with water (100 ml) and extracted with EA (50 ml x 2). The combined organic phase is washed with brine (50 ml), dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 40% EA in hexanes) to give tert-butyl (4-(hydroxymethyl)-5-fluoropyridin-2-yl)(4-methoxybenzyl)carbamate as a colorless oil (5.5 g, 77 %). MS (ESI ⁺ ) m/z 363.2 (M+H) ⁺ , retention time: 1.86 min. (method A). ^1H NMR (400 MHz, CDCl ₃ ) δ 1.40 (s, 9H), 2.48 (s, 3H), 3.76 (s, 3H), 4.62 (m, 2H), 5.14 (s, 2H), 6.80 (d, 2H), 6.84 (s, 1H), 7.20 (d, 2H), 7.40 (s, 1H).

Step 4. Preparation of tert-butyl-[4-(bromomethyl)-5-fluoropyridin-2-yl](4-methoxybenzyl)carbamate

To a solution of tert-butyl-(4-(hydroxymethyl)-5-fluoropyridin-2-yl)(4-methoxybenzyl)carbamate (5.5 g, 15.2 mmol) and carbon tetrabromide (5.9 g, 17.8 mmol ) in DCM (60 ml) at 0°C add triphenylphosphine (4.7 g, 17.9 mmol) in one portion. The reaction mixture is stirred for 20 minutes, the mixture is allowed to warm to RT and the mixture is stirred for 1 hour. The mixture is diluted with DCM (100 ml), extracted with aqueous NaHCO ₃ (50 ml x 1), dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 0-20% EA in hexanes) to give tert-butyl[4-(bromomethyl)-5-fluoropyridin-2-yl](4-methoxybenzyl)carbamate as a white solid (10.6 g, 80%). MS (ESI+) m/z 421.1 (M+H) ⁺ . Retention time: 2.18 min. (method A). ^1H NMR (400 MHz, CDCl ₃ ) δ 1.40 (s, 9H), 2.46 (s, 3H), 3.78 (s, 3H), 4.38 (s, 2H), 5.14 (s, 2H), 6.80 (d, 2H), 6.84 (s, 1H), 7.22 (d, 2H), 7.46 (s, 1H).

Intermediate 12: tert-Butyl[4-(bromomethyl)-5-methylpyridin-2-yl](4-methoxybenzyl)carbamate

Scheme 15

Step 1. Preparation of methyl 2-((tert-butoxycarbonyl)amino)-5-methylpyridine-4-carboxylate

To a solution of methyl 2-amino-5-methylpyridine-4-carboxylate (6 g, 36.1 mmol) in acetone (20 ml) and tert-butyl alcohol (60 ml) add 4-dimethylaminopyridine (220 mg, 1.8 ml) and di-tert-butyl dicarbonate (9.5 g, 43.5 mmol). The mixture is stirred at RT overnight. The reaction mixture was concentrated and the residue was purified by chromatography (elution: 0-30% EA in hexanes) to give methyl 2-((tert-butoxycarbonyl)amino)-5-methylpyridine-4-carboxylate as a white solid (7.7 g , 85%). ^1H NMR (400 MHz, CDCl ₃ ) δ 1.52 (s, 9H), 2.44 (s, 3H), 3.92 (s, 3H), 8.04 (s, 1H), 8.20 (s, 1H), 8.32 (s, 1H).

Step 2. Preparation of methyl 2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-5-methylpyridine-4-carboxylate

To a solution of methyl 2-((tert-butoxycarbonyl)amino)-5-methylpyridine-4-carboxylate (5 g, 18.8 mmol) in DMF (50 ml) add potassium tert-butoxide (3 g, 24.8 mmol ). The mixture is stirred at RT for 5 minutes. and then p-methoxybenzyl chloride (3.82 g, 24.4 mmol) was added. The resulting mixture was stirred at RT for 12 hours. Water (50 ml) was added to the mixture and the mixture was extracted with EA (50 ml x 2). The combined organic phase is washed with brine, dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 0-20% EA in hexane) to give methyl 2-((t-butoxycarbonyl)(4-methoxybenzyl)amino)-5-methylpyridine-4-carboxylate as a white solid (6.1 g, 84%). MS (ESI ⁺ ) m/z 387.2 (M+H) ⁺ , retention time: 2.14 min. (method A).

Step 3. Preparation of tert-butyl-(4-(hydroxymethyl)-5-methylpyridin-2-yl)(4-methoxybenzyl)carbamate

To a solution of methyl 2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-5-methylpyridine-4-carboxylate (6 g, 15.5 mmol) in THF (50 ml) add lithium borohydride (0. 75 g, 34.4 mmol). The mixture is heated and kept at 40°C for 4 hours. The reaction mixture is cooled to 0°C and carefully quenched by slowly adding dropwise an aqueous solution of NaHCO ₃ (20 ml). The mixture is diluted with water (50 ml) and extracted with EA (50 ml x 2). The combined organic phase is washed with brine (50 ml), dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 40% EA in hexanes) to give tert-butyl (4-(hydroxymethyl)-5-methylpyridin-2-yl)(4-methoxybenzyl)carbamate as a colorless oil (4.78 g, 89 %). MS (ESI ⁺ ) m/z 359.2 (M+H) ⁺ , retention time: 1.74 min. (method A).

Step 4. Preparation of tert-butyl-4-(bromomethyl)-5-methylpyridin-2-yl)(4-methoxybenzyl)carbamate

To a solution of tert-butyl-(4-(hydroxymethyl)-5-methylpyridin-2-yl)(4-methoxybenzyl)carbamate (4.78 g, 13.3 mmol) and carbon tetrabromide (5.17 g, 15.6 mmol) in DCM (50 ml) at 0°C add triphenylphosphine (4.1 g, 15.6 mmol) in one portion. The reaction mixture is stirred for 20 minutes, the mixture is allowed to warm to RT and the mixture is stirred for 1 hour. The mixture is diluted with DCM (100 ml), washed with aqueous NaHCO ₃ (50 ml x 1), dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 0-20% EA in hexanes) to give tert-butyl (4-(bromomethyl)-5-methylpyridin-2-yl)(4-methoxybenzyl)carbamate as a white solid (4.2 g , 75%). MS (ESI+) m/z 422.1 (M+H) ⁺ . Retention time: 2.18 min. (method A). ^1H NMR (400 MHz, CDCl ₃ ) δ 1.40 (s, 9H), 2.34 (s, 3H), 3.76 (s, 3H), 4.38 (s, 2H), 5.14 (s, 2H), 6.80 (d, 2H), 7.20 (d, 2H), 7.60 (s, 1H), 8.20 (s, 1H).

Intermediate 13. Preparation of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-1-(tert-butyldimethylsilyl)-4-oxoazetidine -2-carboxylic acid

To a solution of 4-methoxybenzyl-(2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-1-(tert-butyldimethylsilyl)-4- Oxoazetidine-2-carboxylate (1.4 g, 2.1 mmol, prepared as described in WO 2015/120062) in methanol (20 ml) was added with 10% Pd/C (0.2 g). The mixture is hydrogenated at 30 psi (206.8 kPa) for 2 hours. The mixture is filtered through Celite® and the filter cake is washed with methanol. The solvent was removed to give the product as a white solid (0.97 g, 95%). MS (ESI+) 556.2 (M+H) ⁺ . Retention time: 3.64 min. (method B).

Intermediate 14. Preparation of (2S,3R)-1-[(1,1-dimethylethyl)dimethylsilyl]-4-oxo-3-(pyridin-4-ylmethyl)-2-azetidinecarboxylic acid

To a solution of 4-picolyl chloride hydrochloride (10.0 g, 61 mmol) in 100 ml water add NaHCO ₃ (7.7 g, 91.4 mmol) with stirring. The mixture is extracted with TBME (3 x 100 ml). The extracts were combined, washed with brine, dried over MgSO ₄ and concentrated to give 4-picolyl chloride as a colorless oil (7.5 g, 97%), which was used without further purification.

To a solution of (2S)-1-(tert-butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid (prepared as described in Baldwin et al., Tetrahedron, 1990, 46, 4733-4788) (5 g, 21.8 mmol) in THF (80 ml) at -25°C add LDA (2M in THF, 24 ml) dropwise. The resulting mixture was stirred at -15°C for 30 minutes. A solution of 4-picolyl chloride (4 g, 31.3 mmol) in THF (25 ml) was added dropwise to the mixture. The mixture was stirred at -15°C for 2 hours, then the reaction was quenched with aqueous ammonium chloride (20 ml) and EA was added. The organic layer is separated and the aqueous layer is extracted with EA. The combined organic phase is washed with brine, dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: EA) to give the product as a white solid (1.5 g, 15%). MS (ESI+) m/z 321.1 (M+H) ⁺ , retention time: 1.24 min. (method A).

Intermediate 15. Preparation of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylic acid

(S)-1-(tert-Butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid (10.0 g, 43 mmol) was dissolved in THF (160 ml) and cooled to -20°C. The reaction mixture is treated with LDA (2M in THF, 47 ml, 94 mmol) at a temperature in the range of about -10 to -20°C. 4-(bromomethyl)-N,N-bis(4-methoxybenzylpyridin-2-amine (21.2 g, 49 mmol) in THF (80 ml) was then added while maintaining the temperature of the mixture below -10° C. The reaction mixture was stirred for several hours at -15°C, the mixture is allowed to warm to RT and the mixture is stirred for several hours. The reaction mixture is quenched with water (200 ml) and then refluxed for 3 hours. The reaction mixture is cooled to RT and processed for 5 hours. % aqueous solution of tripotassium phosphate (250 ml).The phases are separated and the aqueous layer is extracted with EA (150 ml x 3) to remove impurities. The aqueous phase is acidified to pH 3.1 with 6 N HCl and then extracted with EA (300 ml x 3). The organic phase is dried and concentrated. The residual EA is treated with heptane (250 ml) to obtain a suspension, which is cooled and filtered. The filter cake is transferred to 40 volumes of isopropyl alcohol (400 ml) and refluxed for about 1 hour. CT and insoluble solids are removed by filtration. In the isopropyl alcohol filtrate, the solvent is replaced with heptane (250 ml), causing the product to precipitate. The heptane suspension is cooled to 5-10°C and filtered. The filter cake was dried to give (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylic acid (12 g, 59% ). MS (ESI+) m/z 462.2 (M+H) ⁺ , retention time: 1.23 min. (method A).

Intermediate 16. Preparation of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-1-(tert-butyl(dimethyl)silyl)-4-oxoazetidine -2-carboxylic acid

(S)-1-(tert-Butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid (10 g, 43 mmol) was dissolved in THF (160 ml) and cooled to -20°C. The reaction mixture is treated with LDA (2M in THF, 47 ml, 94 mmol) at a temperature ranging from about -10 to -20°C and then with 4-(bromomethyl)-N,N-bis(4-methoxybenzylpyridin-2-amine ( 21.2 g, 49 mmol) in THF (80 ml), maintaining the temperature of the mixture below -10°C. The reaction mixture is stirred for several hours at -15°C, the mixture is allowed to warm to RT and the mixture is stirred for several hours. The reaction mixture is quenched with water (200 ml) and then acidified with 10% aqueous citric acid to pH 4. The mixture is extracted with EA (100 ml x 3). The combined extracts are dried over MgSO ₄ and the residue is purified by chromatography (elution: 20%-100). %EA in hexane) to obtain the product as a white solid (14.5 g, 59%). MS (ESI+) m/z 576.3 (M+H) ⁺ , retention time: 1.56 min. A).

Intermediate 17. Preparation of (2S,3R)-3-((2-((tert-butoxycarbonyl)(methyl)amino)pyridin-4-yl)methyl)-1-(tert-butyldimethylsilyl)-4-oxoazetidine-2 -carboxylic acid

(S)-1-(tert-Butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid (12.2 g, 53.2 mmol) was dissolved in THF (195 ml) and cooled to -20°C. The reaction mixture was treated with LDA (2M in THF, 60 ml, 120 mmol) at a temperature ranging from about -10 to -20°C and then tert-butyl (4-(bromomethyl)pyridin-2-yl)methylcarbamate (16 g, 53.1 mmol) in THF (80 ml), maintaining the temperature of the mixture below -10 ^o C. The reaction mixture is stirred for several hours at -15°C, the mixture is allowed to warm to RT and the mixture is stirred for several hours. The reaction mixture is quenched with water (200 ml) and then acidified with 10% aqueous citric acid to pH 4. The mixture is extracted with EA (100 ml x 3). The combined organic phase is dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 20%-100% EA in hexane) to give the product as a white solid (14.4 g, 60%). MS (ESI+) m/z 450.2 (M+H) ⁺ , retention time: 1.85 min. (method A).

Intermediate 18. Preparation of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-5-methylpyridin-4-yl)methyl)-1-(tert-butyldimethylsilyl)- 4-oxoazetidine-2-carboxylic acid

(S)-1-(tert-Butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid (2.42 g, 10.6 mmol) was dissolved in THF (36 ml) and cooled to -20°C. The reaction mixture is treated with LDA (2M in THF, 12 ml, 24 mmol) at a temperature ranging from about -10 to -20°C and then tert-butyl-[4-(bromomethyl)-5-methylpyridin-2-yl] ( 4-methoxybenzyl)carbamate (4.44 g, 10.5 mmol) in THF (10 ml), keeping the temperature of the mixture below -10°C. The reaction mixture is stirred for several hours at -15°C, then the mixture is allowed to warm to RT and the mixture is stirred for several hours. The reaction mixture is quenched with water (20 ml) and then acidified with 10% aqueous citric acid to pH 4. The mixture is extracted with EA (30 ml x 3). The combined organic phase is dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 20%-100% EA in hexane) to give the product as a white solid (3.12 g, 53%). MS (ESI+) m/z 570.3 (M+H) ⁺ , retention time: 2.09 min. (method A).

Intermediate 19. Preparation of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-1-(tert-butyldimethylsilyl)- 4-oxoazetidine-2-carboxylic acid

(S)-1-(tert-Butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid (7.1 g, 30.9 mmol) was dissolved in THF (120 ml) and cooled to -20°C. The reaction mixture is treated with LDA (2M in THF, 34 ml, 68 mmol) at a temperature in the range of about -10 to -20°C followed by tert-butyl-[4-(bromomethyl)-6-methylpyridin-2-yl] (4-methoxybenzyl)carbamate (13 g, 30.8 mmol) in THF (20 ml), keeping the temperature of the mixture below -10°C. The reaction mixture is stirred for several hours at -15°C, then the mixture is allowed to warm to RT and the mixture is stirred for several hours. The reaction mixture is quenched with water (100 ml) and then acidified with 10% aqueous citric acid to pH 4. The mixture is extracted with EA (100 ml x 3). The combined organic layer is dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 20%-100% EA in hexane) to give the product as a white solid (10.5 g, 60%). MS (ESI+) m/z 570.3 (M+H) ⁺ , retention time: 2.10 min. (method A).

Intermediate 20. Preparation of (2S,3R)-3-((2-((tert-butoxycarbonyl)(methyl)amino)-6-methylpyridin-4-yl)methyl)-1-(tert-butyldimethylsilyl)-4- oxoazetidine-2-carboxylic acid

(S)-1-(tert-Butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid (4 g, 17.44 mmol) was dissolved in THF (64 ml) and cooled to -20°C. The reaction mixture is treated with LDA (2M in THF, 20 ml, 40 mmol) at a temperature in the range of about -10 to -20°C followed by tert-butyl-(4-(bromomethyl)-6-methylpyridin-2-yl) methyl carbamate (5.5 g, 17.4 mmol) in THF (15 ml), keeping the temperature of the mixture below -10°C. The reaction mixture is stirred for several hours at -15°C, then the mixture is allowed to warm to RT and the mixture is stirred for several hours. The reaction mixture is quenched with water (40 ml) and then acidified with 10% aqueous citric acid to pH 4. The mixture is extracted with EA (60 ml x 3). The combined organic phase is dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 20%-100% EA in hexane) to give the product as a white solid (4.44 g, 55%). MS (ESI+) m/z 464.4 (M+H) ⁺ , retention time: 3.33 min. (method B).

Intermediate 21. Preparation of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-5-fluoropyridin-4-yl)methyl)-1-(tert-butyldimethylsilyl)- 4-oxoazetidine-2-carboxylic acid

(S)-1-(tert-Butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid (2.2 g, 9.6 mmol) was dissolved in THF (36 ml) and cooled to -20°C. The reaction mixture is treated with LDA (2M in THF, 11 ml, 22 mmol) at a temperature ranging from about -10 to -20 °C followed by tert-butyl-[4-(bromomethyl)-5-fluoropyridin-2-yl] (4-methoxybenzyl)carbamate (4.1 g, 9.6 mmol) in THF (10 ml), keeping the temperature of the mixture below -10°C. The reaction mixture is stirred for several hours at -15°C, then the mixture is allowed to warm to RT and the mixture is stirred for several hours. The reaction mixture is quenched with water (20 ml) and then acidified with 10% aqueous citric acid to pH 4. The mixture is extracted with EA (30 ml x 3). The combined organic phase is dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 20%-100% EA in hexane) to give the product as a white solid (2.49 g, 45%). MS (ESI+) m/z 574.4 (M+H) ⁺ , retention time: 3.79 min. (method B).

Intermediate 22. Synthesis of (R)-1-(2,5-dimethylphenyl)propyl isocyanate

Scheme 16

Step 1. Preparation of [N(E),S(S)]-N-[(2,5-dimethylphenyl)methylene]-2-methyl-2-propanesulfonamide

To a solution of 2,5-dimethylbenzaldehyde (5 g, 37.3 mmol) and (S)-(-)-2-methyl-2-propanesulfonamide (5 g, 41.2 mmol) in THF (50 ml) add titanium ethoxide (IV) (17.8 g, 78.0 mmol). The resulting reaction mixture was heated and maintained at 75°C for 2 hours. The mixture is cooled to RT. Brine (50 ml) was added to the mixture, the mixture was stirred vigorously and then filtered through Celite®. The filter is washed with EA (200 ml). The phases are separated and the aqueous layer is extracted with EA (2 x 50 ml). The combined organic phase is dried and concentrated. The residue was purified by chromatography (elution: 4:1 hexane:EA) to give [N(E),S(S)]-N-[(2,5-dimethylphenyl)methylene]-2-methyl-2-propanesulfonamide as colorless oils (8.7 g, 98%). MS (ESI+) 238.1 (M+H) ⁺ , retention time: 2.05 min. (method A).

Step 2. Preparation of [S(S)]-N-((1R)-1-(2,5-dimethylphenyl)propyl)-2-methyl-2-propanesulfonamide

[N(E),S(S)]-N-[(2,5-dimethylphenyl)methylene]-2-methyl-2-propanesulfonamide (8.7 g g, 36.6 mol) dissolved in TBME (50 ml ). The reaction mixture was cooled to -40°C, ethylmagnesium bromide (3M in ether, 24 ml, 74.3 mmol) was added dropwise and the reaction mixture was stirred at -25°C for 4 hours. Add saturated aqueous ammonium chloride followed by EA. The layers are separated and the aqueous layer is extracted with EA (50 ml). The combined organic phase is washed with brine, dried and concentrated. The residue was purified by chromatography (elution: 2:1 hexane:EA) to give [S(S)]-N-((1R)-1-(2,5-dimethylphenyl)propyl)-2-methyl-2-propanesulfonamide as colorless oil (5.2 g, 53%). MS (ESI+) 268.3 (M+H) ⁺ , retention time: 1.87 min. (method A).

Step 3. Preparation of (R)-1-(2,5-dimethylphenyl)propylamine hydrochloride

[S(S)]-N-((1R)-1-(2,5-dimethylphenyl)propyl)-2-methyl-2-propanesulfonamide (5.2 g, 19.4 mmol) dissolved in methanol (20 ml ) and slowly add 4M HCl in 1,4-dioxane (20 ml). The reaction solution is stirred for 60 minutes. The reaction mixture is concentrated to obtain a solid residue. Hexane (20 ml) was added to the solid residue and the resulting suspension was stirred at RT for 60 minutes. The solid was collected by filtration, washed with hexanes and dried to give (R)-1-(2,5-dimethylphenyl)propylamine hydrochloride (2 g, 54%). MS (ESI+) 164.1 (M+H) ⁺ . Retention time: 1.14 min. (method A).

Step 4. Preparation of (R)-1-(2,5-dimethylphenyl)propyl isocyanate

To a suspension of (R)-1-(2,5-dimethylphenyl)propylamine hydrochloride (2 g, 10.0 mmol) in DCM (30 ml) was added a 1N aqueous solution of NaHCO ₃ (34 ml). The mixture is cooled to 0°C and triphosgene (1 g, 3.4 mmol) is added. The reaction mixture is stirred for 15 minutes. at 0°C and for 1 hour at RT. The reaction mixture is extracted twice with DCM. The combined organic phase was dried and concentrated to give (R)-1-(2,5-dimethylphenyl)propyl isocyanate as a colorless oil (1.7 g, 90%), which was used in the next step without further purification. ^1H NMR (400 Hz, CDCl ₃ ) δ 7.21 (s, 1H), 7.05 (d, 1H), 7.01 (d, 1H), 4.74 (t, 1H), 2.36 (s, 3H), 2.31 (s, 3H), 1.82 (m, 2H), 1.05 (t, 3H).

Intermediate 23. Synthesis of (R)-1-(4-fluoro-3-methylphenyl)propyl isocyanate

Scheme 17

Step 1. Preparation of [S(S)]-N-[1-(4-fluoro-3-methylphenyl)propylidene]-2-methylpropane-2-sulfinamide

A solution of 4'-fluoro-3'-methylpropiophenone (5 g, 30 mmol), (S)-(-)-2-methylpropane-2-sulfinamide (4 g, 33 mmol) and titanium (IV) ethoxide (17.4 g, 63 mmol) in THF (100 ml) was heated and kept at 80°C overnight. After cooling, add brine (100 ml) and EA (100 ml). The mixture is stirred at RT for 1 hour. The mixture is filtered through Celite® and the filter is washed with EA. The organic phase is separated, dried and concentrated. The residue is purified by chromatography (elution: 2:1 hexane:EA) to give [S(S)]-N-[1-(4-Fluoro-3-methylphenyl)propylidene]-2-methylpropane-2-sulfinamide as a colorless oil (7.27 g, 90%). MS (ESI+) 270.1 (M+H) ⁺ , retention time: 1.97 min. (method A).

Step 2. Preparation of [S(S),R]-N-[1-(4-fluoro-3-methylphenyl)propyl]-2-methylpropane-2-sulfinamide

To a solution of [S(S)]-N-[1-(4-fluoro-3-methylphenyl)propylidene]-2-methylpropane-2-sulfinamide (7.27 g, 27 mmol) in THF (80 ml) at 0 °C add a solution of tri-sec-butylborohydride (L-selectride, 1M solution in THF, 81 ml, 81 mmol). The reaction mixture is stirred for 3 hours at 0°C. The reaction is quenched with saturated aqueous ammonium chloride (80 ml). The mixture is filtered through Celite® and the filter is washed with EA. The organic phase is separated, dried and concentrated. The residue was purified by chromatography (elution: 1:1 hexane:EA) to give [S(S),R]-N-[1-(4-fluoro-3-methylphenyl)propyl]-2-methylpropane-2-sulfinamide as colorless oil (6.1 g, 84%). MS (ESI+) 272.2 (M+H) ⁺ , retention time: 1.83 min. (method A).

Step 3. Preparation of (R)-1-(4-fluoro-3-methylphenyl)-1-propanamine hydrochloride

To a solution of [S(S),R]-N-[1-(4-fluoro-3-methylphenyl)propyl]-2-methylpropane-2-sulfinamide (6.1 g, 22.5 mmol) in methanol (50 ml) slowly add 4M HCl in 1,4-dioxane (23 ml). The reaction mixture is stirred for 1 hour. The reaction mixture is concentrated to obtain a solid residue. Hexane (50 ml) was added to the solid residue and the resulting suspension was stirred at RT for 1 hour. The solid was collected by filtration, washed with hexanes and dried to give (R)-1-(4-fluoro-3-methylphenyl)-1-propanamine hydrochloride (4.1 g, 90%). MS (ESI+) m/z 168.1 (M+H) ⁺ . Retention time: 1.13 min. (method A).

Step 4. Preparation of (R)-1-(4-fluoro-3-methylphenyl)propyl isocyanate

To a suspension of (R)-1-(4-fluoro-3-methylphenyl)-1-propanamine hydrochloride (1 g, 4.9 mmol) in DCM (30 ml) was added a 1N aqueous solution of NaHCO ₃ (17 ml). The mixture is cooled to 0°C and triphosgene (0.5 g, 1.7 mmol) is added. The reaction mixture is stirred for 15 minutes. at 0°C and for 1 hour at RT. The reaction mixture is extracted twice with DCM. The combined organic phase was dried and concentrated to give (R)-1-(4-fluoro-3-methylphenyl)propyl isocyanate as a colorless oil (0.6 g, 63%), which was used in the next step without further purification. ^1H NMR (400 Hz, CDCl ₃ ) δ 7.10 (m, 2H), 6.98 (m, 1H), 4.48 (t, 1H), 2.29 (s, 3H), 1.85 (m, 2H), 0.98 (t, 3H).

Intermediate 24. Synthesis of 1,5-dihydro-1-methyl-2-(methylamino)-4H-imidazol-4-one hydrochloride

Scheme 18

Step 1. Preparation of tert-butyl-N-(1-methyl-4-oxo-4,5-dihydro-1H-imidazol-2-yl)carbamate

To a solution of 2-imino-1-methyl-4-imidazolidinone (8.2 g, 72.5 mmol) in DMF (30 ml) add di-tert-butyl dicarbonate (17.4 g, 79.7 mmol) at RT. The resulting mixture was stirred at 60°C overnight. The mixture is cooled to RT and quenched with water (30 ml). The mixture is extracted with EA (30 ml x 2). The organic layer is dried and concentrated. The residue is purified by chromatography (elution: 1:1 hexane:EA) to obtain tert-butyl-N-(1-methyl-4-oxo-4,5-dihydro-1H-imidazol-2-yl)carbamate as a white solid (14 g, 90%). MS (ESI+) 214.2 (M+H) ⁺ , retention time: 1.25 min. (method A).

Step 2. Preparation of tert-butyl-N-methyl-N-(1-methyl-4-oxo-4,5-dihydro-1H-imidazol-2-yl)carbamate

To a solution of tert-butyl-N-(1-methyl-4-oxo-4,5-dihydro-1H-imidazol-2-yl)carbamate (14 g, 65.7 mmol) in THF (100 ml) at 0° Sodium hydride (2.9 g, 60% in mineral oil) is added in portions. The mixture is allowed to warm to RT and the mixture is stirred for 30 minutes. Iodomethane (14 g, 95.2 mmol) was slowly added to the reaction mixture. The resulting mixture was stirred at RT overnight. The reaction is quenched by adding saturated aqueous ammonium chloride solution (30 ml). The mixture is extracted with EA (50 ml x 3). The combined organic phases are washed with brine, dried and concentrated. The residue is purified by chromatography (elution: 4:1 hexane:EA) to give tert-butyl-N-methyl-N-(1-methyl-4-oxo-4,5-dihydro-1H-imidazol-2-yl)carbamate in as a colorless oil (3.5 g, 23%). MS (ESI+) 228.2 (M+H) ⁺ , retention time: 1.35 min. (method A).

Step 3. Preparation of 1,5-dihydro-1-methyl-2-(methylamino)-4H-imidazol-4-one hydrochloride

To a solution of tert-butyl-N-methyl-N-(1-methyl-4-oxo-4,5-dihydro-1H-imidazol-2-yl)carbamate (3.5 g, 15.4 mmol) in DCM ( 25 ml) add 4N HCl in dioxane (15 ml). The resulting mixture was stirred at RT overnight. Solvents were removed in vacuo to give 1,5-dihydro-1-methyl-2-(methylamino)-4H-imidazol-4-one hydrochloride as a white solid (1.7 g, 90%). MS (ESI+) 128.0 (M+H) ⁺ , retention time: 0.33 min, (method A).

Intermediate 25. Synthesis of (S)-1-(3-chlorophenyl)propyl isocyanate

Scheme 19

Step 1. Preparation of [N(E),S(S)]-N-[(3-chlorophenyl)methylene]-2-methyl-2-propanesulfonamide

To a solution of 3-chlorobenzaldehyde (58.5 g, 416 mmol) and (S)-(-)-2-methyl-2-propanesulfonamide (53 g, 437 mmol) in THF (500 ml) add titanium (IV) ethoxide ( 190 g, 832 mmol). The resulting reaction mixture was heated and maintained at 75°C for 4 hours. The mixture is cooled to RT. Brine (500 ml) was added to the mixture, the mixture was stirred vigorously and then filtered through Celite®. The filter is washed with EA (600 ml). The phases are separated and the aqueous layer is extracted with EA (2 x 150 ml). The combined organic phase is dried and concentrated. The residue was purified by chromatography (elution: 4:1 hexane:EA) to give [N(E),S(S)]-N-[(3-chlorophenyl)methylene]-2-methyl-2-propanesulfonamide as a colorless oil ( 100 g, 98%). MS (ESI+) 244.1 (M+H) ⁺ , retention time: 2.01 min. (method A).

Step 2. Preparation of [S(S)]-N-((1S)-1-(3-chlorophenyl)propyl)-2-methyl-2-propanesulfonamide

[N(E),S(S)]-N-[(3-chlorophenyl)methylene]-2-methyl-2-propanesulfonamide (100 g, 410 mol) was dissolved in TBME (500 ml). The reaction mixture was cooled to -20°C, ethylmagnesium bromide (3M in ether, 274 ml, 820 mmol) was added dropwise to the mixture and the reaction mixture was stirred at -20°C for 4 hours. A saturated aqueous solution of ammonium chloride and then EA are added to the mixture. The layers are separated and the aqueous layer is extracted with EA (200 ml). The combined organic phase is washed with brine, dried and concentrated. The residue is purified by chromatography, eluting first with 4:1 hexane:EA to give [S(S)]-N-((1S)-1-(3-chlorophenyl)propyl)-2-methyl-2-propanesulfonamide as a colorless oil (65 g, 58%). MS (ESI+) 274.1 (M+H) ⁺ , retention time: 1.85 min. (method A). The column was further eluted with ethyl acetate to provide [S(S)]-N-((1S)-1-(3-chlorophenyl)propyl)-2-methyl-2-propanesulfonamide as a colorless oil (33 g, 29%). MS (ESI+) 274.1 (M+H) ⁺ , retention time: 1.82 min. (method A).

Step 3. Preparation of (S)-1-(3-chlorophenyl)propylamine hydrochloride

[S(S)]-N-((1S)-1-(3-chlorophenyl)propyl)-2-methyl-2-propanesulfonamide (65 g, 238 mmol) was dissolved in methanol (200 ml) and the resulting solution was slowly added add 4M HCl in 1,4-dioxane (240 ml). The reaction solution is stirred for 60 minutes. The reaction mixture is concentrated to obtain a solid residue. Hexane (200 ml) was added to the solid residue and the resulting suspension was stirred at RT for 60 minutes. The solid was collected by filtration, washed with hexanes and dried to give (S)-1-(3-chlorophenyl)propylamine hydrochloride (35 g, 71%). MS (ESI+) 170.1 (M+H) ⁺ , retention time: 1.12 min. (method A).

Step 4. Preparation of (S)-1-(3-chlorophenyl)propyl isocyanate

To a suspension of (S)-1-(3-chlorophenyl)propylamine hydrochloride (9.7 g, 47 mmol) in DCM (150 ml) was added a 1N aqueous solution of NaHCO ₃ (155 ml). The mixture is cooled to 0°C and triphosgene (4.8 g, 16 mmol) is added. The reaction mixture is stirred for 15 minutes at 0°C and for 1 hour at RT. The reaction mixture is extracted twice with DCM. The combined organic phase was dried and concentrated to give (S)-1-(3-chlorophenyl)propyl isocyanate as a colorless oil (7.3 g, 80%), which was used in the next step without further purification. ^1H NMR (400 Hz, _CDCl3 ) δ 7.31 (m, 3H), 7.16 (s, 1H), 4.54 (t, 1H), 1.86 (m, 2H), 0.99 (t, 3H).

Intermediate 26. Synthesis of (R)-1-(3-chlorophenyl)propyl isocyanate

Scheme 20

Step 1. Preparation of [S(S)]-N-[1-(3-chlorophenyl)propylidene]-2-methylpropane-2-sulfinamide

A solution of 3'-chloropropiophenone (75 g, 445 mmol), (S)-(-)-2-methylpropane-2-sulfinamide (57 g, 470 mmol) and titanium(IV) ethoxide (213 g, 934 mmol) in THF (700 ml) is heated and kept at 80°C overnight. After cooling, add brine (700 ml) and EA (700 ml). The mixture is stirred at RT for 1 hour. The mixture is filtered through Celite® and the filter is washed with EA. The organic phase is separated, dried and concentrated. The residue was purified by chromatography (elution: 2:1 hexane:EA) to give [S(S)]-N-[1-(3-chlorophenyl)propylidene]-2-methylpropane-2-sulfinamide as a colorless oil (100 g, 83%). MS (ESI+) 272.1 (M+H) ⁺ , retention time: 2.00 min. (method A).

Step 2. Preparation of [S(S),R]-N-[1-(3-chlorophenyl)propyl]-2-methylpropane-2-sulfinamide

To a solution of [S(S)]-N-[1-(3-chlorophenyl)propylidene]-2-methylpropane-2-sulfinamide (25 g, 92 mmol) in THF (250 ml) at 0°C add a solution of tri- sec-butylborohydride (L-selectride, 1M solution in THF, 185 ml, 185 mmol). The reaction mixture is stirred for 3 hours at 0°C. The reaction mixture is quenched with saturated aqueous ammonium chloride (250 ml). The mixture is filtered through Celite® and the filter is washed with EA. The organic phase is separated, dried and concentrated. The residue was purified by chromatography (elution: 1:1 hexane:EA) to give [S(S),R]-N-[1-(3-chlorophenyl)propyl]-2-methylpropane-2-sulfinamide as a colorless oil (20 .7 g, 82%). MS (ESI+) 274.1 (M+H) ⁺ , retention time: 1.83 min. (method A).

Step 3. Preparation of (R)-1-(3-chlorophenyl)-1-propanamine hydrochloride

To a solution of [S(S),R]-N-[1-(3-chlorophenyl)propyl]-2-methylpropane-2-sulfinamide (20.7 g, 75.8 mmol) in methanol (150 ml) add slowly 4M HCl in 1,4-dioxane (76 ml). The reaction mixture is stirred for 1 hour. The reaction mixture is concentrated to obtain a solid residue. Hexane (150 ml) was added to the solid residue and the resulting suspension was stirred at RT for 1 hour. The solid was collected by filtration, washed with hexanes and dried to give (R)-1-(3-chlorophenyl)-1-propanamine hydrochloride (14 g, 90%). MS (ESI+) m/z 170.1 (M+H) ⁺ , retention time: 1.12 min. (method A).

Step 4. Preparation of (R)-1-(3-chlorophenyl)propyl isocyanate

To a suspension of (R)-1-(3-chlorophenyl)-1-propanamine hydrochloride (38.7 g, 188 mmol) in DCM (300 ml) was added a 1N aqueous solution of NaHCO ₃ (646 ml). The mixture is cooled to 0°C and triphosgene (18.9 g, 63.8 mmol) is added. The reaction mixture is stirred for 15 minutes. at 0°C and for 1 hour at RT. The reaction mixture is extracted twice with DCM. The combined organic phase was dried and concentrated to give (R)-1-(3-chlorophenyl)propyl isocyanate as a colorless oil (29.4 g, 80%), which was used in the next step without further purification. ^1H NMR (400 Hz, CDCl ₃ ) δ 7.32 (m, 3H), 7.20 (m, 1H), 4.54 (t, 1H), 1.82 (m, 2H), 0.98 (t, 3H).

Example 2: Obtaining additional intermediates

The intermediates presented in Tables 2, 3 and 4 below are synthesized as shown in Schemes 6-20 and the experimental procedures of Example 1. In some cases, additional functional group transformations are used. Typically such transformations will be readily apparent to one skilled in the art of organic synthesis. In addition, one skilled in the art of organic synthesis will appreciate that starting materials and reaction conditions can be varied to obtain the desired end products.

Table 2. Additional intermediates

Intermediate Structure Intermediate name LC-MS: retention time (min.) MS (M+H) ⁺ (R)-1-(thiophen-2-yl)propylamine 1.04 142.1 (R)-1-(thiophen-3-yl)propylamine 1.07 142.2 (R)-1-cyclohexylbutylamine 1.12 156.2 Phenyl N-[(R)-cyclohexylcyclopropylmethyl]carbamate 2.09 274.2 Phenyl-N-((R)-1-(6-methylpyridin-3-yl)propyl)carbamate 1.16 271.1 Phenyl-N-((R)-1-cyclopropylpropyl)
carbamate 1.83 220.1 Phenyl-N-((R)-1-(1-methyl-1H-pyrazol-3-yl)propyl)carbamate 1.54 260.1 Phenyl-N-((S)-1-cyclopropyl-2,2,2-trifluoroethyl)carbamate 1.80 260.0

Table 3. Additional intermediates

Intermediate structure Intermediate name ¹H NMR (400 Hz, CDCl₃or DMSO-d6) Phenyl-N-((R)-1-cyclohexyl-2-methylpropyl)carbamate ^1H NMR ( _CDCl3 ) δ 7.38 (m, 2H), 7.20 (m, 1H), 7.17 (m, 2H), 4.70 (br. d, 1H), 3.38 ( m, 1H), 1.92-1.40 (m, 6H), 1.38-0.8 (m, 12H); MS: 276.2 (M+H) ⁺ , retention time: 2.16 min. (R)-1-Cyclohexylbutyl isocyanate ^1H NMR ( _CDCl3 ) δ 0.95 (t, 3H), 1.00-1.28 (m, 4H), 1.38 (m, 2H), 1.40-1.80 (m, 9H ), 3.24 (m, 1H), (R)-1-(3-chlorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 0.98 (t, 3H), 1.82 (m, 2H), 4.54 (t, 1H), 7.20 (m, 1H), 7.32 (m, 3H) (R)-1-(3-methylphenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 0.98 (t, 3H), 1.84 (m, 2H), 2.40 (s, 3H), 4.52 (t, 1H), 7.12 (m, 3H), 7.24 (m, 1H) (R)-1-(3-ethylphenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 0.98 (t, 3H), 1.24 (t, 3H), 1.84 (m, 2H), 2.64 (q, 2H), 4.54 (t, 1H), 7.16 (m, 3H), 7.32 (m, 1H) (R)-1-(3-chlorophenyl)ethyl isocyanate ^1H NMR ( _CDCl3 ) δ 1.60 (d, 3H), 4.78 (q, 1H), 7.20 (m, 1H), 7.34 (m, 3H), (R)-1-Cyclohexylpentyl isocyanate ¹ H NMR (CDCl ₃ ) δ 0.90 (t, 3H), 1.00-1.60 (m, 12H), 1.64 (m, 2H), 1.78 (m, 3H), 3, 24 (m, 1H), (R)-1-(thiophen-2-yl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 1.02 (t, 3H), 1.96 (m, 2H), 4.78 (t, 1H), 7.02 (m, 2H), 7.24 (m , 1H), 1-chloro-3-(isocyanatomethyl)benzene ^1H NMR ( _CDCl3 ) δ 2.50 (s, 2H), 7.20 (m, 1H), 7.36 (m, 3H), N,1-dimethyl-1H-pyrazol-4-amine hydrochloride ^1H NMR (DMSO-d6) δ 2.78 (s, 3H), 3.81 (s, 3H), 4.30 (br., 2H), 7.62 (s, 1H), 8.03 ( s, 1H), N,1-dimethyl-1H-pyrazol-3-amine hydrochloride ^1H NMR (DMSO-d6) δ 2.80 (s, 3H), 3.60 (br., 2H), 3.82 (s, 3H), 6.18 (d, 1H), 7.80 ( d, 1H), N,1,4-trimethyl-1H-pyrazol-5-amine hydrochloride ^1H NMR (DMSO-d6) δ 2.04 (s, 3H), 2.98 (s, 3H), 3.66 (s, 3H), 3.80 (br., 2H), 7.78 ( s, 1H) N,1-dimethyl-1H-imidazol-4-amine hydrochloride ^1H NMR (DMSO-d6) δ 2.62 (s, 3H), 3.69 (s, 3H), 3.80 (br., 2H), 6.60 (s, 1H), 8.50 ( s, 1H) N-methyl-4-isothiazolamine hydrochloride ^1H NMR (DMSO-d6) δ 2.80 (s, 3H), 4.80 (br., 2H), 8.60 (s, 1H), 8.80 (s, 1H) N-methylthiophene-2-amine hydrochloride ^1H NMR (DMSO-d6) δ 2.74 (s, 3H), 4.02 (br., 2H), 6.20 (m, 1H), 6.78 (m, 2H), N-methylthiophene-3-amine hydrochloride ^1H NMR (DMSO-d6) δ 2.80 (s, 3H), 4.06 (br., 2H), 7.56 (m, 1H), 7.64 (m, 2H)

Table 4. Additional intermediates

Intermediate Structure Intermediate name ¹H NMR (400 Hz, CDCl₃or DMSO-d6) (R)-(+)-1-(3,4,5-trifluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 6.94 (m, 2H), 4.54 (t, 1H), 1.84 (m, 2H), 0.99 (t, 3H) (R)-(+)-1-(2,3-difluoro-4-methylphenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.04 (m, 1H), 6.96 (m, 1H), 4.84 (t, 1H), 2.30 (s, 3H), 1.87 (m, 2H), 1.00 (t, 3H) (R)-(+)-1-(2-fluoro-3-methylphenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.21 (m, 1H), 7.14 (m, 1H), 7.06 (m, 1H), 4.87 (t, 1H), 2.30 (s, 3H), 1.88 (m, 2H), 1.01 (t, 3H) (R)-(+)-1-(3,5-difluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 6.85 (m, 2H), 6.72 (m, 1H), 4.57 (t, 1H), 1.86 (m, 2H), 1.00 (t, 3H) (R)-(+)-1-(2,4-difluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.37 (m, 1H), 6.91 (m, 1H), 6.79 (m, 1H), 4.85 (t, 1H), 1.88 (m, 2H), 1.00 (t, 3H) (R)-(+)-1-(3,5-dimethylphenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 6.96 (s, 1H), 6.93 (s, 2H), 4.46 (t, 1H), 2.35 (s, 3H), 2.34 (s, 3H), 1.86 (m, 2H), 1.00 (t, 3H) (R)-(+)-1-(3-fluoro-5-methylphenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 6.89 (m, 1H), 6.83 (m, 1H), 6.80 (s, 1H), 4.50 (t, 1H), 2.37 (s, 3H), 1.84 (m, 2H), 0.99 (t, 3H) (R)-(+)-1-(2,3,4-trimethylphenyl)
propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.20 (d, 1H), 7.08 (d, 1H), 4.84 (t, 1H), 2.33 (s, 3H), 2.27 (s, 3H), 2.24 (s, 3H), 1.85 (m, 2H), 1.06 (t, 3H) (R)-(+)-1-(2,3,4-trifluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.15 (m, 1H), 7.01 (m, 1H), 4.87 (t, 1H), 1.88 (m, 2H), 1.01 (t, 3H) (R)-(+)-1-(2,3-dimethyl-4-fluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.23 (m, 1H), 6.92 (m, 1H), 4.80 (t, 1H), 2.25 (s, 3H), 2.21 (s, 3H), 1.82 (m, 2H), 1.04 (t, 3H) (R)-(+)-1-(2-methyl-3,4-difluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.36 (m, 2H), 4.85 (t, 1H), 2.34 (s, 3H), 1.87 (m, 2H), 1.01 (t, 3H) (R)-(+)-1-(2,3-dimethyl-4-fluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.18 (m, 1H), 6.87 (m, 1H), 4.82 (t, 1H), 2.20 (s, 3H), 2.21 (S, 3H), 1.87 (m, 2H), 1.00 (t, 3H) (R)-(+)-1-(3,5-dimethyl-4-fluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 6.92 (s, 1H), 6.94 (s, 1H), 4.42 (t, 1H), 2.28 (s, 3H), 2.27 (S, 3H), 1.83 (m, 2H), 0.97 (t, 3H) (R)-(+)-1-(3-fluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.32 (m, 1H), 7.09 (m, 1H), 7.01 (m, 1H), 4.56 (t, 1H), 1.86 (m, 2H), 0.99 (t, 3H) (R)-(+)-1-(2-chloro-
5-methylphenyl)
propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.27 (s, 1H), 7.22 (d, 1H), 7.05 (d, 1H), 5.02 (t, 1H), 2.35 (s, 3H), 1.76 (m, 2H), 1.04 (t, 3H) (R)-(+)-1-(3,4-difluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.16 (m, 2H), 7.02 (m, 1H), 4.52 (t, 1H), 1.84 (m, 2H), 0.99 (t, 3H) (R)-(+)-1-(2-fluoro-4-methylphenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.27 (t, 1H), 6.97 (d, 1H), 6.87 (d, 1H), 4.82 (t, 1H), 2.35 (s, 3H), 1.87 (m, 2H), 0.99 (t, 3H) (R)-(+)-1-(2-methyl-4-fluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.27 (t, 1H), 6.98 (d, 1H), 6.87 (d, 1H), 4.83 (t, 1H), 2.35 (s, 3H), 1.87 (m, 2H), 0.99 (t, 3H) (R)-(+)-1-(2,4-dimethylphenyl)
propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.28 (d, 1H), 7.06 (d, 1H), 6.98 (s, 1H), 4.73 (t, 1H), 2.31 (s, 3H), 2.32 (s, 3H), 1.82 (m, 2H), 1.02 (t, 3H) (R)-(+)-1-(2,5-difluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.15 (m, 1H), 6.97 (m, 2H), 4.89 (t, 1H), 1.87 (m, 2H), 1.01 (t, 3H) (R)-(+)-1-(2,3-difluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.17 (m, 1H), 7.12 (m, 2H), 4.91 (t, 1H), 1.90 (m, 2H), 1.02 (t, 3H) (R)-(+)-1-(2,3-dimethylphenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.28 (d, 1H), 7.14 (m, 2H), 4.85 (t, 1H), 2.32 (s, 3H), 2.23 (s, 3H), 1.83 (m, 2H), 1.05 (t, 3H) (R)-(+)-1-(3-fluoro-4-methylphenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.17 (m, 1H), 6.95 (m, 2H), 4.50 (t, 1H), 2.28 (s, 3H), 1.84 (m, 2H), 0.99 (t, 3H) (R)-(+)-1-(4-methylphenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.18 (s, 4H), 4.49 (t, 1H), 2.36 (s, 3H), 1.85 (m, 2H), 0.98 (t, 3H) (R)-(+)-1-(4-chlorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.34 (d, 2H), 7.24 (d, 2H), 4.54 (t, 1H), 1.84 (m, 2H), 0.98 (t, 3H) (R)-(+)-1-(4-fluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.28 (m, 2H), 7.06 (m, 2H), 4.53 (t, 1H), 1.86 (m, 2H), 0.98 (t, 3H) (R)-(+)-1-(2-methyl-4-chlorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.32 (d, 1H), 7.27 (d, 1H), 7.16 (s, 1H), 4.73 (t, 1H), 2.31 (s, 3H), 1.82 (m, 2H), 1.04 (t, 3H) (R)-(+)-1-(3-chloro-4-fluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.36 (m, 1H), 7.16 (m, 2H), 4.54 (t, 1H), 1.84 (m, 2H), 0.99 (t, 3H) (R)-1-(oxan-2-yl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 4.04 (m, 1H), 3.57 (m, 1H), 3.24 (m, 1H), 1.62-1.40 (m, 9H), 0. 99 (t, 3H) (R)-1-(oxan-3-yl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 4.02 (m, 1H), 3.42 (m, 1H), 3.22 (m, 1H), 1.68-1.40 (m, 9H), 0. 99 (t, 3H) (R)-1-(oxan-4-yl)propyl isocyanate ¹ H NMR (CDCl ₃ ) δ 4.01 (m, 1H), 3.38 (m, 1H), 3.22 (m, 1H), 1.68-1.40 (m, 9H), 1, 01 (t, 3H) (R)-(+)-1-(4-trifluoromethoxyphenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.33 (d, 2H), 7.22 (d, 2H), 4.58 (t, 1H), 1.86 (m, 2H), 0.99 (t, 3H) (S)-(-)-1-(3-fluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.32 (m, 1H), 7.09 (m, 1H), 7.01 (m, 1H), 4.56 (t, 1H), 1.86 (m, 2H), 0.99 (t, 3H) (S)-(-)-1-(2,5-difluorophenyl)propyl isocyanate ^1H NMR ( _CDCl3 ) δ 7.15 (m, 1H), 6.97 (m, 2H), 4.89 (t, 1H), 1.87 (m, 2H), 1.01 (t, 3H)

Example 3. (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-phenylpropyl )-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (compound 1-3)

Scheme 21

Step 1. Preparation of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylic acid

(S)-1-(tert-Butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid (10.0 g, 43 mmol) was dissolved in THF (160 ml) and cooled to -20 °C. The reaction mixture is treated with LDA (92M in THF, 47 ml, 94 mmol) at a temperature ranging from about -10 to -20°C and 4-(bromomethyl)-N,N-bis(4-methoxybenzylpyridin-2-amine) is added 21.2 g, 49 mmol) in THF (80 ml), maintaining the temperature of the mixture below -10°C. The reaction mixture is stirred for several hours at -15°C, then the mixture is allowed to warm to RT and the mixture is stirred for several hours. hours. The reaction mixture is quenched with water (200 ml) and then refluxed for 3 hours. The reaction mixture is cooled to RT and treated with 5% aqueous potassium triphosphate (250 ml). The phases are separated and the aqueous layer is extracted with EA (150 ml ×). 3) to remove impurities. The aqueous phase is acidified to pH 3.1 with 6 N HCl and extracted with EA (300 ml x 3). The organic phase is dried over MgSO ₄ and the residual EA is replaced with heptane (250 ml) to obtain a suspension. , which is cooled and filtered. The filter cake is transferred to 40 volumes of isopropyl alcohol (400 ml) and refluxed for about 1 hour. The mixture is cooled to RT and insoluble solids are removed by filtration. In the isopropyl alcohol filtrate, the solvent is replaced with heptane (250 ml), causing the product to precipitate. The suspension is cooled to 5-10°C and filtered. The filter cake was dried to give (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylic acid (12 g, 59% ). MS (ESI+) m/z 462.2 (M+H) ⁺ , retention time: 1.23 min. (method A).

Step 2. Preparation of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-1-(((R)-1-phenylethyl)carbamoyl)-4 -oxoazetidine-2-carboxylic acid

(2S,3R)-3-((2-(Bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylic acid (15 g, 32.5 mmol) dissolved in DCM (150 ml) and treated with DBU (17.3 g, 113.6 mmol) and then (R)-(+)-1-phenylpropyl isocyanate (13.1 g, 81.3 mmol) at RT. The reaction mixture was stirred at RT overnight. The mixture is diluted with DCM (150 ml) and washed with several portions of 10% aqueous citric acid until HPLC detects DBU in the organic phase. The organic phase is dried over MgSO ₄ and concentrated. The residue was purified by chromatography (elution: 20%-100% EA in hexane) to give the product as a white solid (19.2 g, 95%). MS (ESI+) m/z 623.3 (M+H) ⁺ , retention time: 1.55 min. (method A).

Step 3. Preparation of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)- N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

A solution of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-1-(((R)-1-phenylethyl)carbamoyl)-4-oxoazetidin- 2-carboxylic acid (124 mg, 0.2 mmol) in DCM (3 ml) was treated with N,1-dimethyl-1H-pyrazol-3-amine hydrochloride (41 mg, 0.3 mmol) and 2-chloro-1 chloride ,3-dimethylimidazolinium (50 mg, 0.3 mmol). The mixture was stirred at RT and TEA (61 mg, 0.6 mmol) was added. The reaction mixture is stirred overnight; the mixture is then diluted with DCM (20 ml). The mixture is washed with an aqueous solution of NaHCO ₃ , water, dried over MgSO ₄ and concentrated. The oily residue was purified by preparative TLC (2:1 hexane/EA) to give the product as a white solid (92 mg, 65%). MS (ESI+) m/z 716.4 (M+H) ⁺ , retention time: 2.95 min. (method B).

Step 4. Preparation of (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1- phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

Solution of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-( (R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (63 mg, 0.09 mmol) in DCM (3 ml) is treated with triethylsilane (0.1 ml) and cooled to 0° C. Trifluoroacetic acid (1 ml) is added dropwise to the mixture and the reaction mixture is slowly heated to RT. After 21 hours, HPLC analysis (Method A) shows that the starting material has been consumed. The reaction mixture is concentrated and the residue is taken up in 5 ml of DCM. The mixture is washed with an aqueous solution of NaHCO ₃ , water, dried over MgSO ₄ and concentrated. The residue was purified by preparative TLC (elution: EA) to give the title compound as a white solid (21 mg, 50%). MS (ESI+) m/z 476.3 (M+H) ⁺ , retention time: 1.23 min. (method A). ^1H NMR ( _CDCl3 ) δ 0.84 (t, 3H), 1.80 (sq, 2H), 2.80 (m, 1H), 2.84 (m, 1H), 3.24 (s, 3H), 3.56 (m, 1H), 3.84 (s, 3H), 4.40 (s, 1H), 4.60 (m, 2H), 4.76 (m, 1H), 6. 20-6.32 (m, 3H), 6.74 (d, 1H), 7.20-7.40 (m, 6), 7.94 (d, 1H).

Example 4. (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-( 3-chlorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (compound 1-4)

Scheme 22

Step 1. Preparation of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-1-(tert-butyl(dimethyl)silyl)-4-oxoazetidin- 2-carboxylic acid

(S)-1-(tert-butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid (10 g, 43 mmol) was dissolved in THF (160 ml) and cooled to -20°C. The reaction mixture is treated with LDA (2M in THF, 47 ml, 94 mmol) at a temperature in the range of about -10 to -20°C followed by treatment with 4-(bromomethyl)-N,N-bis(4-methoxybenzylpyridin-2-amine (21.2 g, 49 mmol) in THF (80 ml), maintaining the temperature of the mixture below -10°C. The reaction mixture is stirred for several hours at -15°C, then allow the mixture to warm to RT and stir the mixture for several hours. several hours. The reaction mixture is quenched with water (200 ml) and acidified with 10% citric acid to pH 4. The mixture is extracted with EA (100 ml x 3), the organic phase is dried over MgSO ₄ and the residue is concentrated by chromatography (elution: 20% - 100). %EA in hexane) to obtain the product as a white solid (14.5 g, 59%). MS (ESI+) m/z 576.3 (M+H) ⁺ , retention time: 1.56 min. A).

Step 2. Preparation of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-1-(tert-butyl(dimethyl)silyl)-N2-(1 -methyl-1H-pyrazol-5-yl)-N2-methyl-4-oxoazetidine-2-carboxamide

Solution of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-1-(tert-butyl(dimethyl)silyl)-4-oxoazetidine-2-carboxylic acid acid (4 g, 6.9 mmol) in DCM (30 ml) is treated with N,1-dimethyl-1H-pyrazol-5-amine hydrochloride (1.6 g, 10.8 mmol) and 2-chloro-1 chloride, 3-dimethylimidazolinium (1.8 g, 10.6 mmol). The reaction mixture was stirred at RT and TEA (2.5 g, 24.7 mmol) was added to the mixture. The reaction mixture was stirred overnight, then diluted with DCM (20 ml). The mixture is washed with aqueous NaHCO ₃ and water, dried over MgSO ₄ and concentrated. The oily residue was purified by chromatography (elution: 2:1 hexane/EA) to give the product as a white solid (3.2 g, 70%). MS (ESI+) m/z 669.6 (M+H) ⁺ , retention time: 1.77 min. (method A).

Step 3. Preparation of (2S,3R)-3-((2-((bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl) -N2-methyl-4-oxoazetidine-2-carboxamide

To a solution of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-1-(tert-butyl(dimethyl)silyl)-N2-(1-methyl -1H-pyrazol-5-yl)-N2-methyl-4-oxoazetidine-2-carboxamide (3.2 g, 4.8 mmol) in methanol (30 ml) add 0.5 M NH ₄ F in methanol (15 ml) and acetic acid (1.5 ml). The reaction mixture was stirred at RT for 2 hours. The reaction mixture is concentrated and the residue is dissolved in EA (30 ml). The reaction mixture was extracted with aqueous NaHCO ₃ (10 ml) and brine (10 ml), dried over MgSO ₄ and concentrated to give the product as a white solid (2.4 g, 90%). MS (ESI+) 555.3 (M+H) ⁺ , retention time: 1.28 min. (method A).

Step 4. Preparation of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)- N1-((R)-1-(3-chlorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

Solution of (2S,3R)-3-((2-((bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N2- Methyl 4-oxoazetidine-2-carboxamide (1.2 g, 2.2 mmol) in DCM (10 ml) was treated with TEA (1.1 g, 10.8 mmol) and (R)-1-( 3-chlorophenyl)propyl isocyanate (1.1 g, 5.6 mmol) and stir the reaction mixture at 50°C until HPLC analysis (Method B) shows the reaction is complete. Pour the reaction mixture into 40 ml of water and. the aqueous phase is extracted with DCM (3 x 15 ml). The combined organic phase is dried over MgSO ₄ and concentrated. The colorless viscous oily residue is purified by chromatography (elution: 2:1 hexane/EA) to give the title compound as a white foam (0. 9 g, 60%). MS (ESI+) m/z 750.3 (M+H) ⁺ , retention time: 3.17 min (method B).

Step 5. Preparation of (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (3-chlorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

Solution of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-( (R)-1-(3-chlorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (0.9 g, 1.2 mmol) in DCM (10 ml) treated with triethylsilane (1 ml) and cool to 0°C. Trifluoroacetic acid (2 ml) was added dropwise to the mixture and the reaction mixture was slowly heated to RT. After 21 hours, HPLC analysis shows that the starting material has been consumed. The reaction mixture is concentrated and the residue is taken up in 5 ml of DCM. The mixture is washed with aqueous NaHCO ₃ and water, dried over MgSO ₄ and concentrated. The residue was purified by preparative TLC (10% methanol in DCM) to give the title compound as a white solid (340 mg, 55%). MS (ESI+) m/z 510.2 (M+H) ⁺ , retention time: 1.25 min. (method A).

Example 5. (2S,3R)-3-((2-dimethylaminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1-phenylpropyl )-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (compound 1-5)

Scheme 23

Step 1. Preparation of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)- N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

A solution of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-1-(((R)-1-phenylethyl)carbamoyl)-4-oxoazetidin- 2-carboxylic acid (product of Example 3, Step 2, 100 mg, 0.16 mmol) in DCM (5 ml) was treated with N,1-dimethyl-1H-pyrazol-4-amine dihydrochloride (50 mg, 0.27 mmol) and 2-chloro-1,3-dimethylimidazolinium chloride (50 mg, 0.3 mmol). The mixture was stirred at RT and TEA (100 mg, 0.98 mmol) was added to the mixture. The reaction mixture was stirred overnight and then diluted with DCM (20 ml). The mixture is washed with an aqueous solution of NaHCO ₃ , water, dried over MgSO ₄ and concentrated. The oily residue was purified by preparative TLC (2:1 hexane/EA) to give the product as a white solid (69 mg, 60%). MS (ESI+) m/z 716.4 (M+H) ⁺ , retention time: 2.95 min. (method B).

Step 2. Preparation of (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

Solution of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-( (R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (69 mg, 0.09 mmol) in DCM (3 ml) is treated with triethylsilane (0.1 ml) and cooled to 0° C. Trifluoroacetic acid (1 ml) is added dropwise and the reaction mixture is slowly heated to RT. After 21 hours, HPLC analysis shows that the starting material has been consumed. The reaction mixture is concentrated and the residue is taken up in 5 ml of DCM. The mixture is washed with an aqueous solution of NaHCO ₃ , water, dried over MgSO ₄ and concentrated. The residue was purified by preparative TLC (elution: EA) to give the title compound as a white solid (38 mg, 83%). MS (ESI+) m/z 476.3 (M+H) ⁺ , retention time: 1.23 min. (method A), ^1H NMR ( _CDCl3 ) δ 0.86 (t, 3H), 1.80 (q, 2H), 2.10 (m, 2H), 2.62 (m, 1H), 2 .84 (m,1H), 3.16 (s, 3H), 3.60 (m, 1H), 3.88 (s, 3H), 4.12 (s, 1H), 4.60 (m, 2H), 4.74 (m, 1H), 6.12 (s, 1H), 6.30 (d, 1H), 6.70 (d, 1H), 7.20-7.40 (m, 5H ), 7.96 (d, 1H).

Step 3. Preparation of (2S,3R)-3-((2-dimethylaminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

Acetic acid (40 mg, 0.7 mmol) is added to the solution of (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl) -N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (36 mg, 0.08 mmol), formaldehyde (160 mg, 37% aqueous solution, 2.0 mmol ) and NaBH ₃ CN (42 mg, 0.7 mmol) in ACN (5 ml) and water (1 ml) at RT. The reaction mixture is stirred for 2 days. The reaction mixture is diluted with water (10 ml) and made alkaline to pH 4 with 1M aqueous NaOH and then extracted with EA (10 ml x 2). The organic layer was washed with brine (5 mL), dried over MgSO ₄ and concentrated. The residue was purified by preparative TLC (10% methanol in DCM) to give (2S,3R)-3-((2-dimethylaminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)- N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide as a white solid (25 mg, 66%). MS (ESI+) m/z 504.2 (M+H) ⁺ , retention time: 1.22 min. (method A).

Example 6. (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-cyclohexyl -2,2,2-trifluoroethyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (compound 1-6)

Solution of (2S,3R)-3-((2-((bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N2- Methyl 4-oxoazetidine-2-carboxamide (product of Example 4, Step 3, 100 mg, 0.18 mmol) in DCM (10 ml) was treated with TEA (100 mg, 0.98 mmol). 1-cyclohexyl-2,2,2-trifluoroethyl]carbamate (100 mg, 0.48 mmol) was added and the reaction mixture was stirred at 50°C until HPLC analysis (Method B) indicated the reaction was complete. the mixture is poured into 40 ml water and the aqueous phase is extracted with DCM (3 x 15 ml). The combined organic phase is dried over MgSO ₄ and concentrated. The colorless viscous oily residue is purified by chromatography (elution: 2:1 hexane/EA) to give (2S,3R). )-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1 -cyclohexyl-2,2,2-trifluoroethyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide as white foam (105 mg, 70%) MS (ESI+) m/z 762.6 (M+). H) ⁺ , retention time: 1.90 min (method A).

Solution of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-( (R)-1-cyclohexyl-2,2,2-trifluoroethyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (105 mg, 0.14 mmol) in DCM (3 ml) was treated with triethylsilane (0. 1 ml) and cool to 0°C. Trifluoroacetic acid (1 ml) was added dropwise to the mixture and the reaction mixture was slowly heated to RT. After 21 hours, HPLC analysis shows that the starting material has been consumed. The reaction mixture is concentrated and the residue is taken up in 5 ml of DCM. The mixture is washed with aqueous NaHCO ₃ and water, dried over MgSO ₄ and concentrated. The residue was purified by preparative TLC (elution: EA) to give the title compound as a white solid (36 mg, 50%). MS (ESI+) m/z 522.4 (M+H) ⁺ , retention time: 1.28 min. (method A).

Example 7. (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-( 3-chlorophenyl)propyl)-N1-methyl-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (compound 1-7)

Scheme 24

Step 1. Preparation of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)- N1-((R)-1-(3-chlorophenyl)propyl)-N1-methyl-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

To a solution of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1- ((R)-1-(3-chlorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (prepared as in Example 4, Step 4, 300 mg, 0.4 mmol) in DMF (4 ml) add sodium hydride (19 mg, 60% in mineral oil, 0.48 mmol). The mixture is stirred for 5 minutes. Methyl iodide (68 mg, 0.48 mmol) was added to the mixture. The mixture is stirred at RT for 30 minutes. The reaction mixture is quenched with a few drops of water. The reaction mixture is diluted with EA (10 ml) and water (10 ml). The organic layer was washed with brine, dried over MgSO ₄ and concentrated. The crude product was purified by preparative TLC (elution: EA) to give the title compound as a white solid (91 mg, 30%). MS (ESI+) m/z 764.6 (M+H) ⁺ , retention time: 1.78 min. (method A).

Step 2. Preparation of (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1- (3-chlorophenyl)propyl)-N1-methyl-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

Solution of (2S,3R)-3-((2-(bis(4-methoxybenzyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-( (R)-1-(3-chlorophenyl)propyl)-N1-methyl-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (91 mg, 0.12 mmol) in DCM (2 ml) treated with triethylsilane (0 .1 ml) and cooled to 0°C. The reaction mixture is treated with trifluoroacetic acid (0.5 ml), adding it dropwise with stirring, and slowly warming the mixture to RT. After 21 hours, HPLC analysis (Method A) shows that the starting material has been consumed. The reaction mixture was concentrated and the residue was taken up in DCM (15 ml). The mixture is washed with saturated aqueous NaHCO ₃ and water, dried over MgSO ₄ and concentrated. The crude product was purified by preparative TLC (elution: EA) to give the title compound as a white solid (31 mg, 50%). MS (ESI+) m/z 524.4 (M+H) ⁺ , retention time: 1.25 min. (method A).

Example 8. Synthesis of (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R )-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

Scheme 25

Step 1. Preparation of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-1-(tert-butyldimethylsilyl)-4 -oxoazetidine-2-carboxylic acid

(S)-1-(tert-Butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid (125 g, 545 mmol) was dissolved in THF (2 L) and cooled to -20 ^° C. The reaction mixture was treated with LDA (2M in THF , 600 ml, 102 mol), maintaining the temperature in the range from -10 to -20°C. The resulting gel-like mixture was stirred at -15°C for 30 minutes. Then add tert-butyl[4-(bromomethyl)-6-methylpyridin-2-yl](4-methoxybenzyl)carbamate (230 g, 546 mmol) in THF (500 ml), maintaining the temperature of the mixture below -10°C. The reaction mixture is stirred at -10°C for 2 hours, then the mixture is cooled to -20°C, quenched with water (200 ml) and then acidified to pH 4 with 10% aqueous citric acid, maintaining the temperature of the mixture below 10°C. The mixture is extracted with EA (500 ml x 3). The combined organic layer is dried and concentrated. The residue is purified by chromatography (eluting with a gradient of 20%-50% EA/hexane) to give (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4 -yl)methyl)-1-(tert-butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid as a white solid (155 g, 50%). MS (ESI+) m/z 570.3 (M+H) ⁺ , retention time: 2.10 min. (method A).

Step 2. Preparation of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-1-(tert-butyldimethylsilyl)-N2 -(1-methyl-1H-imidazol-2-yl)-N2-methyl-4-oxoazetidine-2-carboxamide

To a solution of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-1-(tert-butyldimethylsilyl)-4-oxoazetidine -2-carboxylic acid (146 g, 256 mmol) in DMF (500 ml) add HATU (146 g, 384 mmol). The mixture is stirred for 10 minutes. at room temperature. N,1-dimethyl-1H-imidazol-2-amine (37 g, 333 mmol) was added to the mixture and the resulting solution was stirred for 10 minutes. and then diisopropylethylamine (99.5 g, 770 mmol) was added. The mixture was stirred at RT for 2 hours and then brine (600 ml) was added. The mixture is extracted with EA (500 ml x 3). The combined organic phase is washed with water (300 ml) and concentrated. The residue is purified by chromatography (eluting with a gradient of 20%-50% EA/hexane) to give (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4 -yl)methyl)-1-(tert-butyldimethylsilyl)-N2-(1-methyl-1H-imidazol-2-yl)-N2-methyl-4-oxoazetidine-2-carboxamide as a semi-solid (85 g, 50 %). MS (ESI+) m/z 663.7 (M+H) ⁺ , retention time: 2.19 min. (method A).

Step 3. Preparation of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H- imidazol-2-yl)-N2-methyl-4-oxoazetidine-2-carboxamide

To a solution of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-1-(tert-butyldimethylsilyl)-N2-( 1-methyl-1H-imidazol-2-yl)-N2-methyl-4-oxoazetidine-2-carboxamide (118 g, 178 mmol) in methanol (300 ml) add NH ₄ F (12 g, 324 mmol) and acetic acid acid (21 g, 333 mmol). The reaction mixture was stirred at RT for 2 hours. The reaction mixture is concentrated and the residue is dissolved in EA (500 ml). The mixture was washed with saturated aqueous NaHCO ₃ (200 ml x 2) and brine (200 ml), dried and concentrated to give (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl) )amino)-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N2-methyl-4-oxoazetidin-2-carboxamide as a white solid (85 g , 87%). MS (ESI+) 549.4 (M+H) ⁺ , retention time: 1.56 min. (method A).

Step 4. Preparation of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H- imidazol-2-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

Solution of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2 -yl)-N2-methyl-4-oxoazetidine-2-carboxamide (119 g, 217 mmol) in DCM (500 ml) was treated with TEA (110 g, 1.09 mol). (R)-(+)-1-phenylpropyl isocyanate (87.4 g, 542 mmol) was added to the mixture, and the reaction mixture was stirred at 50°C for 16 hours. The reaction mixture is poured into water (400 ml) and the phases are separated. The aqueous phase is extracted with DCM (3 x 150 ml). The combined organic phase is dried and concentrated. The colorless viscous oily residue is purified by chromatography (elution: 2:1 hexane/EA) to give (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4- yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide as a white solid substances (123 g, 80%). MS (ESI+) 710.6 (M+H) ⁺ , retention time: 2.11 min. (method A).

Step 5. Preparation of (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R )-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

To a solution of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol- 2-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (140 g, 197 mmol) in trifluoroacetic acid (800 ml) slowly add triethylsilane (137, 6 g, 1.18 mol). The resulting mixture was stirred at RT overnight. Solvents are removed in vacuo. The residue is treated with saturated aqueous NaHCO ₃ solution (500 ml). The mixture is extracted with DCM (300 ml x 2). The organic phase is dried and concentrated. The residue was purified by chromatography (elution: 5% methanol/DCM) to give an oil, which was triturated with hexane/EA (8:1) to give (2S,3R)-3-((2-amino-6-methylpyridine-4 -yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide as a solid white matter (82 g, 85%). MS (ESI+) 490.4 (M+H) ⁺ , retention time: 2.11 min. (method B). ^1H NMR (400 Hz, CDCl ₃ ) δ 0.82 (t, 3H), 1.80 (m, 2H), 2.32 (s, 3H), 2.76 (m, 1H), 2.96 (m, 1H), 3.20 (s, 3H), 3.60 (s, 3H), 3.72 (m, 2H), 4.68 (m, 1H), 5.30 (m, 2H) , 6.20 (s, 1H), 6.24 (s, 1H), 6.68 (d, 1H), 6.94 (s, 1H), 7.04 (s, 1H), 7.28 ( m, 3H), 7.34 (m, 2H).

Example 9. Synthesis of (2S,3R)-3-((1-oxide-2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1 -((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

To a solution of (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)- 1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (25 mg, 0.05 mmol, product of Example 1) in DCM (5 ml) add meta-chloroperoxybenzoic acid (18 mg, 0.08 mmol ). The mixture is stirred for 1 hour at RT. The mixture is diluted with DCM (10 ml). The mixture is washed with saturated aqueous NaHCO ₃ solution and water, dried and concentrated. The residue was purified by preparative TLC (10% methanol/DCM) to give (2S,3R)-3-((1-oxide-2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H -imidazol-2-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide as a white solid (15 mg, 58%). MS (ESI+) m/z 510.2 (M+H) ⁺ , retention time: 1.25 min. (method A).

Example 10. Synthesis of (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(5-oxo-1-methyl-4,5-dihydro-1H-imidazol- 2-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

Scheme 26

Step 1. Preparation of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylic acid

To a solution of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-1-(tert-butyldimethylsilyl)-4-oxoazetidine -2-carboxylic acid (10 g, 17.6 mmol) in methanol (50 ml) add NH ₄ F (1.3 g, 35 mmol) and acetic acid (2.1 g, 33.3 mmol). The reaction mixture was stirred at RT for 2 hours, concentrated and the residue was dissolved in DCM (100 ml). The mixture was washed with water (20 ml) and brine (20 ml), dried and concentrated to give (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6- Methylpyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylic acid as a white solid (7 g, 88%). MS (ESI+) m/z 456.2 (M+H) ⁺ , retention time: 1.55 min. (method A).

Step 2. Preparation of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-1-(((R)-1 -phenylethyl)carbamoyl)-4-oxoazetidine-2-carboxylic acid

(2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylic acid (7.7 g , 16.9 mmol) dissolved in DCM (100 ml) and treated with DBU (9 g, 59.1 mmol) and then (R)-(+)-1-phenylpropylisocyanate (6.9 g, 42.6 mmol) with CT. The reaction mixture was stirred at RT overnight. The mixture is diluted with DCM (50 ml) and the mixture is washed with several portions of 10% aqueous citric acid until HPLC analysis (Method A) shows no DBU. The organic phase is dried and concentrated. The residue is purified by chromatography (gradient elution: 20% EA/hexane to 100% EA) to give (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridine -4-yl)methyl)-1-(((R)-1-phenylethyl)carbamoyl)-4-oxoazetidine-2-carboxylic acid as white solid (9.3 g, 90%), MS (ESI+) m/z 617.3 (M+H) ⁺ , retention time: 3.66 min. (method B).

Step 3. Preparation of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-N2-(5-oxo-1- methyl 4,5-dihydro-1H-imidazol-2-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidin-1,2-dicarboxamide

To a solution of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-1-(((R)-1-phenylethyl )carbamoyl)-4-oxoazetidine-2-carboxylic acid (100 mg, 0.16 mmol) in DMF (2 ml) was added HATU (123 mg, 0.32 mmol). The mixture is stirred for 10 minutes. with CT. 1-Methyl-2-methylamino-4,5-dihydroimidazol-5-one hydrochloride (83 mg, 0.5 mmol, prepared as described in J. Org. Chem., 1968, 33, 552) was added and the resulting solution was stirred in for 10 minutes. Diisopropylethylamine (120 mg, 0.9 mmol) was added. The mixture was stirred at RT for 2 hours and then brine (6 mL) was added. The mixture is extracted with EA (5 ml x 3). The combined organic phase is washed with water (3 ml) and concentrated. The residue is purified by chromatography (eluting with a gradient of 20%-50% EA/hexane) to give (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4 -yl)methyl)-N2-(5-oxo-1-methyl-4,5-dihydro-1H-imidazol-2-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4- oxoazetidine-1,2-dicarboxamide as a semi-solid (58 mg, 50%). MS (ESI+) m/z 726.7 (M+H) ⁺ , retention time: 2.05 min. (method A).

Step 4. Preparation of (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(5-oxo-1-methyl-4,5-dihydro-1H-imidazol- 2-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide

Solution of (2S,3R)-3-((2-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-methylpyridin-4-yl)methyl)-N2-(5-oxo-1-methyl-4 ,5-dihydro-1H-imidazol-2-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (58 mg, 0.08 mmol) in DCM (3 ml) is treated with triethylsilane (0.1 ml) and cooled to 0°C. Trifluoroacetic acid (1 ml) was added dropwise to the mixture and the reaction mixture was slowly heated to RT. After 21 hours, HPLC analysis (Method A) shows that the starting material has been consumed. The reaction mixture is concentrated and the residue is taken up in 5 ml of DCM. The mixture is washed with saturated aqueous NaHCO ₃ solution and water, dried and concentrated. The residue was purified by preparative TLC (elution: EA) to give (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(5-oxo-1-methyl-4,5 -dihydro-1H-imidazol-2-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide as a white solid (20 mg, 50%). MS (ESI+) m/z 506.5 (M+H) ⁺ , retention time: 1.19 min. (method A).

Example 11: Additional Connection Examples

The compounds presented in Tables 5, 6 and 7 below were synthesized in accordance with the methods shown in Schemes 1-5 and 21-26 and the procedures of Examples 3-10.

Table 5. Additional examples of compounds obtained

Connection structure Connection name LC-MS retention time (min.) LC-MS
(M+H) ⁺
1-8 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(4-thiazolyl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1 ,2-dicarboxamide 1.25 479.4
1-9 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(thiophen-3-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine -1,2-dicarboxamide 1.25 478.4
1-10 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(thiophen-2-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine -1,2-dicarboxamide 1.28 478.3
1-11 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1-(3-chlorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 510.3
1-12 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(5-thiazolyl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1 ,2-dicarboxamide 1.23 479.2
1-13 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-phenylpropyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 476.5
1-14 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(3-methylphenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 490.4
1-15 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (3-chlorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 524.4
1-16 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1-(3-ethylphenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 504.4 1-17 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(4-thiazolyl)-N1-((R)-1-(3-chlorophenyl)propyl)-N2-methyl- 4-oxoazetidine-1,2-dicarboxamide 1.28 513.3
1-18 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1-(6-methylpyridine -3-yl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.06 491.3
1-19 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(4-isothiazolyl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1 ,2-dicarboxamide 1.25 479.3
1-20 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(6-methylpyridine -3-yl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.05 491.3
1-21 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(3-isothiazolyl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1 ,2-dicarboxamide 1.25 479.3
1-22 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1- (3-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 504.4
1-23 (2S,3R)-3-((2-(((tert-butoxy)carbonyl)amino)pyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1- ((R)-1-(3-chlorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.78 610.4
1-24 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(4-thiazolyl)-N1-((R)-1-(3-methylphenyl)propyl)-N2-methyl- 4-oxoazetidine-1,2-dicarboxamide 1.26 493.3
1-25 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-4-yl)-N1-((R)-1-(3-chlorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 510.3
1-26 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((3-chlorophenyl)methyl)-N2- methyl 4-oxoazetidine-1,2-dicarboxamide 1.20 482.3
1-27 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((3-chlorophenyl)methyl)-N2- methyl 4-oxoazetidine-1,2-dicarboxamide 1.20 482.3
1-28 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(thiophen-2-yl)-N1-((R)-1-(3-chlorophenyl)propyl)-N2- methyl 4-oxoazetidine-1,2-dicarboxamide 1.30 512.3
1-29 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(thiophen-2-yl)-N1-((3-chlorophenyl)methyl)-N2-methyl-4-oxoazetidin- 1,2-dicarboxamide 1.26 484.3
1-30 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((S)-1-cyclopropyl-2, 2,2-trifluoroethyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 480.3
1-31 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((S)-1-phenyl-2, 2,2-trifluoroethyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 516.4
1-32 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((S)-1-cyclohexyl-2, 2,2-trifluoroethyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.29 522.4
1-33 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((S)-1-(3-chlorophenyl )ethyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 496.4
1-34 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-4-yl)-N1-((3-fluorophenyl)methyl)-N2- methyl 4-oxoazetidine-1,2-dicarboxamide 1.17 466.3
1-35 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(3-chlorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 510.3
1-36 (2S,3R)-3-((2-methylaminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-(3-chlorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 524.4
1-37 (2S,3R)-3-((2-methylaminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(3-chlorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 524.4
1-38 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(3-chlorophenyl )-2,2,2-trifluoroethyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 550.4
1-39 (2S,3R)-3-((2-ethylaminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(3-chlorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.28 538.5
1-40 (2S,3R)-3-((2-methylaminopyridin-4-yl)methyl)-N2-(thiophen-2-yl)-N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine -1,2-dicarboxamide 1.27 492.4
1-41 (2S,3R)-3-((2-methylaminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(3-chlorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 524.4
1-42 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(1-methyl -1H-pyrazol-3-yl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.13 480.4
1-43 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (3-chlorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 524.4
1-44 (2S,3R)-3-((2-methylaminopyridin-4-yl)methyl)-N2-(thiophen-2-yl)-N1-((R)-1-(3-chlorophenyl)propyl)-N2- methyl 4-oxoazetidine-1,2-dicarboxamide 1.31 526.4
1-45 (2S,3R)-3-((2-methylamino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 504.4
1-46 (2S,3R)-3-((2-methylamino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (3-chlorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 538.4
1-47 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-cyclohexylpropyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 482.5
1-48 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1- phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 490.4
1-49 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(thiophen-2-yl)-N1-((R)-1-(1-methyl-1H-pyrazole-3 -yl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.18 482.4
1-50 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-cyclohexylethyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 468.4
1-51 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 490.4
1-52 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-4-yl)-N1-((R)-1- phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.18 490.4
1-53 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(thiophene-2 -yl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.20 482.4
1-54 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(thiophene-3 -yl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.21 482.4
1-55 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-cyclohexylpropyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 482.4
1-56 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-cyclohexyl-2, 2,2-trifluoroethyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 522.1
1-57 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-cyclohexyl-2- methylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.28 496.1
1-58 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-cyclohexyl-cyclopropylmethyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide 1.28 494.4
1-60 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-(3-chlorophenyl )propyl)-N1-methyl-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 524.4
1-61 (2S,3R)-3-((2-methylamino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 504.5
1-62 (2S,3R)-3-((2-methylamino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- cyclohexylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.29 510.5
1-63 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-cyclohexylpropyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 482.4
1-64 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-methyl-N2-methyl-4-oxoazetidin-1 ,2-dicarboxamide 1.04 372.3
1-65 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(3-chlorophenyl )propyl)-N1-methyl-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 524.4
1-66 (2S,3R)-3-((2-methylamino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 504.5
1-67 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- cyclohexylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 496.5
1-68 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-cyclohexylbutyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide 1.29 496.5
1-69 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1-cyclohexylpropyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 482.5
1-70 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-4-yl)-N1-((R)-1-cyclohexyl-2, 2,2-trifluoroethyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 522.2
1-71 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-4-yl)-N1-((R)-1-cyclohexylpropyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 482.3
1-72 (2S,3R)-3-((2-methylamino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-4-yl)-N1-((R)-1- phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.21 504.2
1-73 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-cyclohexylpentyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide 1.33 510.3
1-74 (2S,3R)-3-((2-dimethylaminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-phenylpropyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 504.3
1-75 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- cyclohexylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 496.3
1-76 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-4-yl)-N1-((R)-1- cyclohexylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 496.3
1-77 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- cyclohexyl-2,2,2-trifluoroethyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 536.3
1-78 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1,4-dimethyl-1H-pyrazol-5-yl)-N1-((R)-1-phenylpropyl) -N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 490.2
1-79 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(3-fluorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.21 494.2
1-80 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(3-methylphenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 490.2
1-81 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(6-methylpyridine -3-yl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 0.59 491.2
1-82 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (6-methylpyridin-3-yl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.05 505.2
1-83 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(4-methylphenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 510.2
1-84 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2,5 -difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 512.2
1-85 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-phenyl-2- methylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 490.3
1-86 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(3-fluoro -2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 508.2
1-87 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(4-methylphenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.20 490.2

Table 6. Additional examples of compounds obtained

Connection structures Connection name ¹ H NMR (CDCl ₃ )
LC-MS(M+H) ⁺
LC-MS retention time (min.)
1-88 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1-phenylpropyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide ^1H NMR ( _CDCl3 ) δ 0.86 (t, 3H), 1.80 (q, 2H), 2.10 (m, 2H), 2.62 (m, 1H), 2.84 (m, 1H), 3.16 (s, 3H), 3.60 (m, 1H), 3.88 (s, 3H), 4.12 (s, 1H), 4.60 (m, 2H), 4. 74 (m, 1H), 6.12 (s, 1H), 6.30 (d, 1H), 6.70 (d, 1H), 7.20-7.40 (m, 5H), 7.96 (d, 1H)
(M+H) ⁺ : 476.3
Time hold: 1.23
1-89 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1- (3-chlorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide ^1H NMR ( _CDCl3 ) δ 0.88 (t, 3H), 1.80 (sq, 2H), 2.30 (s, 3H), 2.72 (m, 1H), 2.84 (m, 1H), 3.16 (s, 3H), 3.54 (m, 1H), 3.70 (m, 1H), 3.84 (s, 3H), 4.20 (s, 1H), 4. 60 (m, 3H, 6.02 (s, 1H), 6.20 (s, 1H), 6.24 (d, 1H), 6.78 (d, 1H), 7.10-7.40 ( m, 4H)
(M+H) ⁺ : 524.2
Time hold: 1.26 min,
1-90 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-4-yl)-N1-((R)-1-phenylpropyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide ^1H NMR ( _CDCl3 ) δ 0.88 (t, 3H), 1.82 (sq, 2H), 2.84 (m, 2H), 3.16 (s, 3H), 3.60 (m, 1H), 3.72 (s, 3H), 4.50 (s, 1H), 4.78 (m, 3H, 5.04 (m, 2H), 6.22 (s, 1H), 6.38 (d, 1H), 6.80 (d, 1H), 7.04 (s, 1H), 7.20-7.40 (m, 5H), 7.84 (d, 1H),
(M+H) ⁺ : 476.3
Time hold: 1.19 min.
1-91 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(3-chlorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide ^1H NMR ( _CDCl3 ) δ 0.88 (t, 3H), 1.80 (sq, 2H), 2.80 (m, 1H), 2.90 (m, 1H), 3.16 (s, 3H), 3.58 (m, 1H), 3.86 (s, 3H), 4.40 (s, 1H), 4.60-4.78 (m, 3H), 6.20 (d, 1H ), 6.22 (d, 1H), 6.38 (d, 1H), 6.76 (d, 1H, 7.10-7.40 (m, 4H), 7.94 (d, 1H),
(M+H) ⁺ : 510.2
Time hold: 1.26 min,
1-92 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-phenylpropyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide ^1H NMR ( _CDCl3 ) δ 0.84 (t, 3H), 1.80 (sq, 2H), 2.80 (m, 2H), 3.16 (s, 3H), 3.40 (m, 1H), 3.52 (m, 1H), 3.80 (s, 3H), 4.56 (m, 2H), 4.74 (m, 1H), 5.80 (s, 1H), 5. 84 (s, 1H), 6.24 (d, 1H), 6.62 (d, 1H), 7.20-7.40 (m, 5H), 7.54 (d, 1H), 7.86 (d, 1H)
(M+H) ⁺ : 477.2
Time hold: 1.22 min. 1-93 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((S)-1-(3-chlorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide (M+H) ⁺ : 510.2
Time hold: 1.23 min,

Table 7. Additional examples of compounds obtained

Connection structures Connection name LC-MS retention time (min.)
(method A) LC-MS
(M+H) ⁺
1-94 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(4-chlorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 510.1
1-95 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2,6 -dimethylpyridin-3-yl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 0.89 505.2
1-96 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((S)-1-(2,6 -dimethylpyridin-3-yl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.02 505.2
1-97 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(4-trifluoromethoxyphenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 560.5
1-98 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(4-fluorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 494.1
1-99 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2,5 -dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 504.4
1-100 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(3,4 -dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 504.5 1-101 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2-chloro -5-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 524.4
1-102 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2-fluoro -5-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 508.4
1-103 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(3-chloro -4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 524.4
1-104 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(3-chloro -2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 524.4
1-105 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (4-chlorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 524.4
1-106 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (4-fluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.21 508.4
1-107 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 504.0
1-108 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (2,5-dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 518.1
1-109 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (3,4-dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 518.5
1-110 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (5-chloro-3-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 538.4
1-111 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (2-chloro-5-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 538.4
1-112 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (2-fluoro-5-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 522.4
1-113 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (3-chloro-4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 538.4
1-114 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2,4 -dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 504.3
1-115 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2-chloro -3-fluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 528.0
1-116 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2-chloro -4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 524.3
1-117 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(3-methyl -4-fluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 508.4
1-118 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(4-chloro -2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 524.4
1-119 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2-fluoro -4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.28 508.8
1-120 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (3-fluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 508.4
1-121 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(oxan-2 -yl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.17 484.0
1-122 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(oxan-4 -yl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.13 484.4
1-123 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(2,5 -difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 512.0
1-124 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(4-methylphenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 490.4
1-125 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(2-fluoro -5-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 508.4
1-126 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(oxan-3 -yl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.14 484.4
1-127 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(3,4 -dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 504.4
1-128 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(2-methyl -3-fluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 508.4
1-129 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(3-chloro -4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 524.3
1-130 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(2-methylphenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 490.4
1-131 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(4-oxo-1-methyl-4,5-dihydro-1H-imidazol-2-yl)-N1-( (R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 492.4
1-132 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(2,5 -dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 504.4
1-133 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(2-fluorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.21 494.3
1-134 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(2,3 -difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 512.4
1-135 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(5-fluoro -2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 508.4
1-136 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(3-fluoro -4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 508.4
1-137. (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1- (2,5-difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 526.4
1-138 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(3-chloro -2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 524.5
1-139 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(4-oxo-1-methyl-4,5-dihydro-1H-imidazol-2-yl) -N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 506.5
1-140 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(2,3 -dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 504.5
1-141 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(5-chloro -2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 524.4
1-142 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1- (4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 504.1
1-143 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1- (2-fluoro-5-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 522.5
1-144 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1- (2,5-difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 526.5
1-145 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1- (3,4-dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 5,18,5
1-146 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1- (2,3-difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 526.5
1-147 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1- (3-chloro-4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.28 538.5
1-148 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(5-oxo-1-methyl-4,5-dihydro-1H-imidazol-2-yl) -N1-((R)-1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.19 506.5
1-149 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (2,3-dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 518.6
1-150 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (3-fluoro-2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 522.5
1-151 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (2,3-difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 526.5
1-152 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (3,4-dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 518.5
1-153 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (4-fluoro-3-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 522.5
1-154 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 504.5
1-155 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (3-fluoro-4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 522.5
1-156 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (4-fluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 508.5
1-157 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (4-chlorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 524.5
1-158 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (3,4-difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 526.5
1-159 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (4-fluoro-2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 522.5
1-160 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (2-fluoro-4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 522.6
1-161 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (2,4-dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.28 518.6
1-162 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (5-fluoro-2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 522.5
1-163 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (2,5-difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 526.5
1-164. (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (2-fluoro-5-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.21 522.5
1-165 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(4-methyl-4H-1,2,4-triazol-3-yl)-N1-((R)- 1-phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.20 477.5
1-166 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1- (2,5-dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 518.6
1-167 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1,3,4-oxadiazol-2-yl)-N1-((R)-1-phenylpropyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 464.4
1-168 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1,3,4-thiadiazol-2-yl)-N1-((R)-1-phenylpropyl)-N2 -methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 480.4
1-169 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (4-fluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 508.6
1-170 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (4-chlorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 524.6
1-171 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (4-fluoro-3-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 522.7
1-172 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (3-fluoro-4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 522.6
1-173 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (2,3-dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.29 518.7
1-174 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (2,3-difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 526.6
1-175 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (3,4-dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.28 518.7
1-176 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (2,5-dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 518.7
1-177 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (2-fluoro-5-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 522.7
1-178 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (5-fluoro-2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 522.7
1-179 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (2,4-dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.28 518.7
1-180 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (4-fluoro-2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 522.7
1-181 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 504.6
1-182 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (2,5-difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 526.6
1-183 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (2-fluoro-4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 522.7
1-184 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (3-fluoro-5-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 522.7
1-185 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (3,5-dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.28 518.7
1-186 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1,3,4-thiadiazol-2-yl)-N1-((R)-1- phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 494.5
1-187 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1,3,4-oxadiazol-2-yl)-N1-((R)-1- phenylpropyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 478.6
1-188 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (2,4-difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 526.7
1-189 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (3,5-difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 526.7
1-190 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (2-fluoro-3-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 522.7
1-191 (2S,3R)-3-((2-amino-6-methylpyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1- (3-fluoro-2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 522.7
1-192 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2,3 -difluoro-4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 526.7
1-193 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(3,4 ,5-trifluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 530.7
1-194 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2,4 -difluoro-3-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 526.7
1-195 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(3,5 -dimethyl-4-fluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 522.7
1-196 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2,4 -dimethyl-3-fluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 522.5
1-197 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(3,4 ,5-trifluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 530.3
1-198 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(2,4 -difluoro-3-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 526.3
1-199 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1-(3,4 ,5-trifluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 530.3
1-200 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-(2,4 -difluoro-3-methylphenyl)propyl)-N1-methyl-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 526.4
1-201 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(2,3 -difluoro-4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 526.3
1-202 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-(2,3 -difluoro-4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 526.3
1-203 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-3-yl)-N1-((R)-1-(3,5 -dimethyl-4-fluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.27 522.4
1-204 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-(3,4 ,5-trifluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 530.4
1-205 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-(3,5 -dimethyl-4-fluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 522.4
1-206 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1-(3,5 -dimethyl-4-fluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 522.4
1-207 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1-(2,4 -difluoro-3-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 526.4
1-208 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-4-yl)-N1-((R)-1-(2,3 -difluoro-4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 526.4
1-209 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2,3 ,4-trimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 518.4
1-210 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2,3 ,4-trifluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 530.3 1-211 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2,3 -dimethyl-4-fluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.25 522.3
1-212 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(3,4 -difluoro-2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 526.2
1-213 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-(2,3 -difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 512.2
1-214 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-(4-fluoro -3-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 508.2
1-215 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-(3,4 -difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 512.3
1-216 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-(3-fluoro -4-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.24 508.3
1-217 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-(2,5 -difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 512.2
1-218 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((R)-1-(2-fluoro -3,4-dimethylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.26 522.3
1-219 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-pyrazol-5-yl)-N1-((R)-1-(5-fluoro -2-methylphenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.23 508.3
1-220 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((S)-1-(3-fluorophenyl )propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.21 494.2
1-221 (2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(1-methyl-1H-imidazol-2-yl)-N1-((S)-1-(2,5 -difluorophenyl)propyl)-N2-methyl-4-oxoazetidine-1,2-dicarboxamide 1.22 512.2

Example 12: Determination of Dose-Response Relationships for Protease Inhibitors

Materials:

Assay buffer: 20 mM Hepes, pH 7.4; 150 mM NaCl; 0.02% Tween 20

Compounds: stock solutions 10 mM in DMSO

Substrate: 20 mM Glp-Pro-Arg-AMC, 25 mg/2.3 ml H ₂ O (store at +4°C) [factor XIa, thrombin and trypsin]

20 mM Pro-Phe-Arg-AMC (Bachem I-1295), 25 mg/2.2 ml H ₂ O

or Boc-Leu-Gly-Arg-AMC (Bachem I-1105), 25 mg/2.07 ml H ₂ O

(store at +4°C) [factor Xa]

Enzyme: Factor Xia, 0.25 mM in 50% glycerol (20 μg/ml)

Trypsin: 0.2 mM in 50% glycerol (4.8 μg/ml)

Thrombin: 0.2 mM in 50% glycerol (7.34 μg/ml)

Factor Xa: 0.2 mM in 50% glycerol (9.2 μg/ml)

Stock solutions are aliquoted (~100 µl/aliquot) and stored at -20°C

Methods:

1. Substrate is diluted to 100 mM in assay buffer (30 µl/6 ml). The enzyme is diluted to 0.5 nM immediately before use (12 µl/6 ml for factor XIa; 15 µl/6 ml for all other enzymes).

2. Pipette 50 µl of substrate into each well of a 96-well (12 × 8) microtiter plate. (Column 1 is used as a control for 100% activity and no compound is added to it, and column 12 is a control and no enzyme is added to it.) Column 12 is added with an additional 46 µl.

3. Using a pipette, add 4 µl of each compound to the appropriate well of column 2 of the plate (compounds with unknown activity are analyzed three times, the standard is analyzed twice). The final concentration of the compound will be 1/50 of the original.

4. Consistently dilute the compound solution twofold by mixing the sample in column 2, withdrawing 50 μl and transferring to the next well (column 3), mixing, withdrawing and transferring to column 4, etc. to column 11. After mixing in column 11, withdraw 50 µl and discard.

5. Pipette 50 µl of buffer into column 12. The reaction is initiated by adding 50 μl of enzyme solution to each well of columns 1-11 as quickly as possible.

6. The plate is read in a spectrophotometer (SpectraMax) at 30°C, where each well is read every 60 seconds for 30 minutes. For factor Xa assays, each well is read every 1 min. within 60 min.

7. For factor XIa assays, compounds are tested twice at three different initial concentrations: 20, 2, and 0.2 μM; 1:10, 1:100 and 1:1000 dilution of the stock solution at 10 mM or at 2, 0.2 and 0.02 μM; 1:100, 1:1000 and 1:10,000 dilution of the original solution of 10 mM. Data for each concentration are combined to generate graphs and fit the data.

8. The data obtained is used to determine IC ₅₀ and calculate K _on .

Example 13. Metabolic stability of canine hepatocytes

The procedure described below is used to determine the stability of test compounds in hepatocytes of Beagle dogs in a 96-well plate format. Test compounds, including control, imipramine and 7-ethoxycoumarin, were dissolved at 10 mM in DMSO, followed by an additional 5-fold dilution with DMSO to 2 mM. Pooled cryopreserved hepatocytes from beagle dogs are thawed and transferred to complete medium. The cells are gently centrifuged (700 rpm for 5 min) and the pellets are washed with Krebs-Henseleit buffer (KHB). The cells are resuspended, counted using trypan blue and adjusted to 1 million cells/ml in KHB buffer. Samples of 2 mM stock solutions are diluted 10-fold with acetonitrile, followed by a 50-fold dilution with KHB buffer. 200 µl of the resulting compound solutions are transferred to 96-well polypropylene assay plates, an equal volume of hepatocyte solution is added and mixed to initiate the reaction (final assay concentrations are 2 µM compound and 500,000 cells/ml). Assay plates are incubated at 37°C in 5% CO ₂ with gentle mixing. Aliquates of 50 μl are withdrawn at several time points up to 180 minutes and quenched with 6 volumes of chilled acetonitrile (containing 250 ng/ml carbutamide and chrysin as internal standards) to stop the protein precipitation reaction. The cooled samples are kept on ice. Samples of the slaked mixture are centrifuged at 2000 × g (3100 rpm) for 10 minutes at 4°C. 50 μL aliquots of supernatants were removed and diluted with 100 μL Milli-Q water to reduce the % organic content prior to bioanalysis by LC-MS/MS. The peak areas corresponding to the test compound are determined. The remaining compound is calculated by comparing the peak area at each time point with zero time point. Data are analyzed and results are obtained using Microsoft Excel 2010.

Example 14 Factor XIa Inhibition Assays

The ability of the compounds of the present invention to inhibit factor XIa is assessed by determining the concentration of inhibitor that results in a 50% reduction in enzyme activity (IC ₅₀ ) using purified enzyme. Potential factor XIa inhibitors are evaluated using the following assay.

PyroGlu-Pro-Arg-7-methylaminocourin (AMC), available from CPC Scientific, Inc., is applied to a pyro-Glu-Pro-Arg-pNA (S-2366) support, available from Diapharma Group, Inc. (Columbus, Ohio), in which the p-nitroanaline group is replaced with 7-methylaminocoumarin (AMC).

The final substrate concentration in the assay is 50 µM and the final enzyme concentration is 0.25 nM. Inhibitors are tested by serial dilution over an appropriate concentration range to obtain a dose-response curve and determine the IC ₅₀ value of the inhibitors. The assay mixture is read every minute for 30 minutes to obtain progress curves. The plates were read in a Spectramax m5 Multimode Plate Reader (Molecular Devices LLC, Sunnyvale, CA). The dose-response curves correspond to Equation 1 below, where A is the maximum inhibition, B is the minimum inhibition, C is the IC _50, and D is the Hill coefficient.

Y = [(AB) / (1 + (X/C) ^D )] + B (Equation 1)

Table 8. Efficiency, selectivity and stability of the compounds of the examples presented in Table 1

Connection number hFXIa IC50 (nM) Ratio: FXa/FXIa Ratio: thrombin/FXIa Ratio: trypsin/FXIa Stability of canine hepatocytes
(t _1/2 : min.) 1-3 A I H G N 1-4 A I I G L 1-5 D J J J O 1-6 B J J J O 1-7 B J J J O 1-8 A H G G N 1-9 A H H G O 1-10 A H H G K 1-11 A H H G O 1-12 A H H G O 1-13 A I I G M 1-14 B J J J O 1-15 B H G G N 1-16 A H G G O 1-17 A H H G L 1-18 B H G G M 1-19 A H G G O 1-20 A J J J O 1-21 B J J J O 1-22 A H H G M 1-23 D J J J O 1-24 A J J J O 1-25 A H G G O 1-26 B H G G O 1-27 B J J J O 1-28 A G H G O 1-29 B J J J O 1-30 B J J J O 1-31 B H G G O 1-32 A I H G K 1-33 A H H G O 1-34 C J J J O 1-35 A H H G L 1-36 B J J J O 1-37 B H H G O 1-38 A H G G O 1-39 B J J J O 1-40 B J J J O 1-41 B I H G L 1-42 C J J J O 1-43 A I H G M 1-44 A H H G O 1-45 B I H G M 1-46 A I H G L 1-47 A I H G L 1-48 A I H G N 1-49 B J J J O 1-50 B I H G L 1-51 A I H G N 1-52 B H G G O 1-53 B J J J O 1-54 B J J J O 1-55 A I H G L 1-56 A I H G O 1-57 B I H G K 1-58 A I H G K 1-60 B J J J O 1-61 B J J J O 1-62 B I I G K 1-63 A I I G K 1-64 C J J J O 1-65 B J J J O 1-66 B J J J O 1-67 B I I G L 1-68 B J J J O 1-69 B H G G O 1-70 A I G G O 1-71 A I G G O 1-72 B H G G O 1-73 B J J J O 1-74 D J J J O 1-75 B J J J O 1-76 B J J J O 1-77 B J J J O 1-78 B J J J O 1-79 A I H G O 1-80 A I H G M 1-81 B I H G M 1-82 B J J J O 1-83 B I H G M 1-84 A I H G O 1-85 B J J J O 1-86 C J J J O 1-87 A I H G O 1-88 A H G G N 1-89 A I H G N 1-90 A I H G M 1-91 A H H G L 1-92 A I H G M 1-93 B J J J O 1-94 A H H G O 1-95 B J J J O 1-96 C J J J O 1-97 B H H G O 1-98 A I H G O 1-99 A H H G O 1-100 A H H G O 1-101 A H H G O 1-102 A I H G O 1-103 A H H G O 1-104 A H H G O 1-105 B I H G O 1-106 A I H G O 1-107 B I H G O 1-108 B J J J O 1-109 A H H G O 1-110 A H H G O 1-111 A H H G O 1-112 B I H G O 1-113 A H H G O 1-114 A H H G O 1-115 A J J J O 1-116 A J J J O 1-117 A J J J O 1-118 A J J J O 1-119 A J J J O 1-120 A I H G N 1-121 B J J J O 1-122 B J J J O 1-123 A I H G O 1-124 A J J J O 1-125 A I H G O 1-126 B J J J O 1-127 A H H G O 1-128 A H H G O 1-129 A H H G O 1-130 A J J J O 1-131 B J J J O 1-132 A J J J O 1-133 B J J J O 1-134 A I H G O 1-135 A J J J O 1-136 A H H G O 1-137 A I H G N 1-138 A J J J O 1-139 B J J J O 1-140 A J J J O 1-141 A J J J O 1-142 A J J J O 1-143 A I H G O 1-144 A I H G O 1-145 A J J J O 1-146 A I H G O 1-147 A J J J O 1-148 C J J J O 1-149 B J J J O 1-150 A J J J O 1-151 A I I G O 1-152 A I I G L 1-153 A J J J O 1-154 A J J J O 1-155 A I I G M 1-156 A J J J O 1-157 B J J J O 1-158 A I I G M 1-159 A J J J O 1-160 A J J J O 1-161 A J J J O 1-162 A I I G O 1-163 A I I G O 1-164 A J J J O 1-165 B J J J O 1-166 A J J J O 1-167 A J J J O 1-168 A J J J O 1-169 B J J J O 1-170 B J J J O 1-171 B J J J O 1-172 A J J J O 1-173 B J J J O 1-174 B J J J O 1-175 A J J J O 1-176 B J J J O 1-177 B J J J O 1-178 A J J J O 1-179 A J J J O 1-180 B J J J O 1-181 B J J J O 1-182 A I H G O 1-183 A J J J O 1-184 A I H G O 1-185 A I H G O 1-186 A J J J O 1-187 A J J J O 1-188 B J J J O 1-189 B J J J O 1-190 A J J J O 1-191 B J J J O 1-192 A J J J O 1-193 A I H G O 1-194 A I H H O 1-195 A J J J O 1-196 A J J J O 1-197 A I I H O 1-198 A I H H O 1-199 A I H H O 1-200 A I I H O 1-201 A J J J O 1-202 A J J J O 1-203 A J J J O 1-204 A J J J O 1-205 A J J J O 1-206 A J J J O 1-207 A J J J O 1-208 A J J J O 1-209 A J J J O 1-210 A J J J O 1-211 A J J J O 1-212 A J J J O 1-213 A J J J O 1-214 A J J J O 1-215 A J J J O 1-216 A J J J O 1-217 A J J J O 1-218 A J J J O 1-219 A J J J O 1-220 B J J J O 1-221 C J J J O

Refer to Table 8: FXIa and hFXIa stand for human factor XIa and human factor XIa, respectively.

Efficiency: “A” means <10 nM, “B” means 10-100 nM, “C” means 100-1000 nM, “D” means >1000 nM, and “E” means no data available or not determined.

Selectivity: “F” means < 1, “G” means 1-100, “H” means 101-1000; “I” means >500, and “J” means the data is not available or has not been defined.

Stability of canine hepatocytes: “K” means 0-60 min.; “L” means 61-120 minutes; “M” means 121-240 minutes; “N” means > 240 min; “O” means the data is not available or has not been defined.

In Table 8, the factor XIa enzyme inhibition activities of compounds 1-79 and compounds 1-84 were erroneously assigned to compound 1-220 and compound 1-221, respectively, in USSN 62/627435, to which the present application claims priority. The factor XIa enzyme inhibition values of compound 1-79, compound 1-84, compound 1-220 and compound 1-221 in the present invention are correct.

Example 15 Colon Epithelial Cancer Cell Permeability Assay (Caco-2)

To determine the ability of a test compound to cross-link Caco-2 cells (colon epithelial cancer cells derived from colorectal adenocarcinoma). Cells are seeded at a density of 1 × 105 cells/ ^cm2 in 96-well MultiscreenTM microplates (Millipore). Permeability analysis is carried out with cells 21-25 days after seeding. Cells are typically used for 15 consecutive passages in culture.

This analysis can be performed in an apical to besolateral (A-B) direction or in a basolateral to apical (B-A) direction. Prepare a solution of the test compound at a concentration of 10 μM in HBSS-MES (pH 6.5) or HBSS-HEPES (pH 7.4) with a final DMSO concentration of 1%. The working solution is then centrifuged and the supernatant is added to the donor side. Assay plates are incubated at 37°C with gentle shaking for 60 minutes. or 40 min. for A-B or B-A analysis, respectively. Aliquots of samples are taken from the donor side at time zero and at the end point, and from the acceptor side at the end point.

The reference compounds included in each assay are propranolol (highly permeable), labetalol (moderately permeable), ranitidine (poorly permeable), and colchicine (a P-glycoprotein substrate).

Samples are analyzed by HPLC-MS/MS using selected reaction monitoring. The HPLC system consists of a binary LC pump with autosampler, C-18 column and gradient.

The apparent permeability coefficient (P _app ) of the test compound (Equation 2) and its recovery (Equation 3) are calculated as follows:

(Equation 2)

(Equation 3)

where A is the surface area of the cell monolayer (0.11 cm ² ), C is the concentration of the test compound, represented as peak area, D is donor, and R is acceptor. “0”, “mid” and “end” indicate the time at the zero, mid and end point of incubation, respectively. Δt - incubation time. V is the volume of the donor and acceptor.

Low recovery indicates that the test compound is lost during analysis. This is most likely due to nonspecific binding or degradation.

Example 16 Determination of solubility

To determine the aqueous solubility of the test compound in phosphate buffered saline solution (PBS - NaCl 137 mM, KCl 2.7 mM, Na ₂ HPO ₄ 8.1 mM, KH ₂ PO ₄ 1.5 mM, pH 7.4) in the format 96-well plate using HPLC-UV/VIS analysis technique. From a 10 mM stock solution in DMSO, prepare a 200 μM solution of the test compound in PBS. The final DMSO concentration is 2%. The PBS buffer sample is mixed thoroughly followed by incubation at room temperature for 24 hours. At the end of the incubation, PBS buffer samples were centrifuged and the supernatants analyzed by HPLC. The aqueous solubility (μM) of a test compound in PBS is determined by comparing the area of the main peak in the calibration standard (200 μM) with the corresponding peak area of each PBS sample. The concentration range of the assay is approximately 0.5 µM to 200 µM. The reference compounds used in each assay are metoprolol, rifampicin, ketoconazole, phenytoin, haloperidol, simvastatin, diethylstilbestrol, and tamoxifen, ranked from completely soluble (200 μM) to poorly soluble (< 1 μM).

Example 17 Plasma Protein Binding Assays

The following procedure is used to determine plasma protein binding of a test compound in pooled human plasma (mixed sex) by equilibrated dialysis in a 96-well plate format. Phosphate-buffered solution (pH 7.4) is loaded into the dialysate cell, and plasma containing the test compound at a concentration of 10 μM is loaded into the side of the sample. Once loaded, samples are capped and incubated for 4 hours at 37°C. After incubation, a sample is removed from each well, diluted with acetonitrile/buffer and centrifuged. Supernatants were analyzed by HPLC-MS/MS. The analysis peak areas obtained by HPLC-MS/MS are used to calculate protein binding according to the following equation (Equation 4):

(Equation 4)

Where

Area _p is the area of the analyte peak in the protein matrix

Area _b - peak area of the analyte in the buffer

The amount determined in the plasma cell includes both free and bound drug, while on the buffer side only free drug is present; the differences are used to calculate the percentage of bound plasma protein. Each assay tests three reference compounds: acebutolol, quinidine, and warfarin.

Example 18: Determination of bioavailability in dogs

To assess bioavailability in the dog's body, the following technique is used. The test compounds are formulated with vehicles compatible with intravenous and oral dosing. The test compounds are administered to male Beagle dogs in the form of intravenous boluses at a dose of 1 mg/mg (1 mg/ml in a volume of 1 ml/kg) or through an oral tube at a dose of 5 mg/kg (1 mg/ml in a volume of 5 ml/kg) . Blood samples are collected through the contralateral (for intravenous bolus) jugular vein or another available vein at various time points after dosing and up to 24 hours after dosing. The resulting plasma samples (EDTA) were centrifuged (2200 × g for 10 minutes at 5°C) and stored frozen (70°C) until analyzed for the parent compound by LC/MS/MS. Pharmacokinetic parameters are estimated from concentration-time data by standard non-compartmental methods using standard analysis software. By comparing the area under the curve (AUC) for IV and PO administration, estimates of oral bioavailability are obtained.

Table 9. Caco-2 cell permeability (A-B and B-A), aqueous solubility, human plasma protein binding (PPB), and bioavailability in the dog of the example compounds presented in Table 1.

Connection number Permeability Caco-2 (A-B) Permeability Caco-2 (B-A) Aqueous solubility (µM in PBS) PPB (%) Bioavailability in dogs (%) 1-3 P R W Z FF 1-4 Q Q W Z FF 1-8 P R W Z GG 1-9 Q Q X CC GG 1-10 Q Q W A.A. E.E. 1-11 P Q X CC GG 1-13 P Q W Z FF 1-15 P Q W Z GG 1-17 Q Q W A.A. GG 1-18 P P W Y GG 1-22 P Q W Z GG 1-32 Q Q W A.A. GG 1-35 P R W Z GG 1-41 P R W Z GG 1-43 P Q W Z GG 1-45 Q R W Z E.E. 1-46 Q Q W A.A. GG 1-47 P R W A.A. GG 1-50 P R W Z GG 1-51 P R W Z FF 1-55 P R W Z GG 1-57 Q Q V BB GG 1-58 Q Q W A.A. GG 1-62 Q Q W Z GG 1-63 Q R W Z GG 1-67 Q R W Z GG 1-79 P Q W Z GG 1-80 P R W Z GG 1-81 P P W Y DD 1-83 P S W A.A. GG 1-84 P Q W Z GG 1-88 P Q W Z GG 1-89 P Q W Z FF 1-90 P Q W Z GG 1-91 P Q W Z FF 1-92 P R W Z FF 1-120 P Q W Z GG 1-137 P Q W A.A. GG 1-152 P Q W Z GG 1-155 P Q W Z GG 1-158 P Q W A.A. GG

For Table 9: Caco-2 permeability (P _app ): “P” means ≤ 10 × 10 ^-6 cm/sec; “Q” means 11-30 × 10 ^-6 cm/sec; “R” means 31-50 × 10 ^-6 cm/sec; “S” means > 50 × 10 ^-6 cm/sec; “T” means the data is not available or has not been defined.

Aqueous solubility: “U” means ≤ 10 mM; “V” means 10-100 mM; “W” means >100 mM; “X” means the data is not available or has not been identified.

PPB: “Y” means >50%; "Z" means 50-90%; “AA” means 91-95%; “BB” means >95%; “CC” means the data is not available or has not been defined.

Bioavailability in the dog: “DD” means ≤ 10%; “EE” means 11-30%; “FF” > 30%; “GG” means the data is not available or has not been identified.

Claims (55)

1. Compound of formula (II) or a pharmaceutically acceptable salt thereof, where R ¹ represents NH ₂ ; R ^a represents C _1-6 alkyl; R ^b represents hydrogen or C _1-6 alkyl; R ² represents pyrazolyl or imidazolyl, wherein the pyrazolyl or imidazolyl is optionally substituted with one or two independent C1-6 alkyls; R ³ represents C _1-6 alkyl; R ⁴ is phenyl optionally substituted with one, two or three independent substituents selected from halogen or C _1-6 alkyl; R ⁵ is hydrogen, C _1-6 alkyl, C _1-6 haloalkyl, C _3-6 cycloalkyl ^Rx is missing; m is 0; And n is 1. 2. A compound according to claim 1, wherein R ² represents pyrazolyl or imidazolyl, wherein pyrazolyl or imidazolyl is substituted with one methyl. 3. A compound according to claim 1 or 2, where R ³ represents methyl. 4. A compound according to any one of claims 1 to 3, wherein R ⁴ is phenyl, optionally substituted with one, two or three independent substituents selected from methyl. 5. A compound according to any one of claims 1 to 4, where R ⁵ represents hydrogen, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH ₂ CH ₃ , -CH(CH ₃ ) ₂ , -CH ₂ CH ₂ CH ₂ CH ₃ , -CF ₃ or unsubstituted cyclopropyl. 6. A compound according to claim 1, wherein the compound is a compound of formula (I-a) 7. A compound according to claim 1, wherein the compound is a compound of formula (I-c), formula (I-d), formula (I-e), formula (I-f), formula (I-g), formula (I-i) or formula (I-j): 8. Compound formula: 9. A pharmaceutical composition having the property of inhibiting factor XIa or kallikrein, comprising an effective amount of a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. 10. The pharmaceutical composition according to claim 9, which is a liquid preparation or a solid preparation. 11. A method of treating deep vein thrombosis by inhibiting factor XIa, comprising administering to a patient who has suffered an ischemic attack an effective amount of a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt or composition thereof according to claim 9 or 10. 12. A method of preventing deep vein thrombosis by inhibiting factor XIa in a patient in need thereof, comprising administering to the patient an effective amount of a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt or composition thereof according to claim 9 or 10. 13. A method of inhibiting factor XIa in a patient, comprising administering to the patient who has suffered from ischemia an effective amount of a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof or a composition according to claim 9 or 10. 14. A method of treating a thromboembolic consequence or complication by inhibiting factor XIa in a patient in need thereof, comprising administering to the patient an effective amount of a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt or composition thereof according to claim 9 or 10. 15 . A method of treating a thromboembolic complication by inhibiting factor XIa in a patient in need thereof, comprising administering to the patient an effective amount of a compound of any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof or a composition of claim 9 or 10, wherein the patient is exposed to an artificial surface . 16. A method of reducing the risk of a thromboembolic complication by inhibiting factor XIa in a patient in need thereof, comprising administering to the patient an effective amount of a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt or composition thereof according to claim 9 or 10, in which the patient is subjected to exposure to artificial surfaces. 17. A method of reducing the risk of a thromboembolic consequence or complication by inhibiting factor XIa in a patient in need thereof, comprising administering to the patient an effective amount of a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt or composition thereof according to claim 9 or 10. 18. The method according to claim 15 or 17, wherein the thromboembolic consequence or complication is associated with intervention in peripheral blood vessels, hemodialysis, catheter ablation, cerebrovascular intervention, organ transplantation, surgery, trans-catheter aortic valve implantation, large diameter intervention used for aneurysm treatment, percutaneous coronary intervention, or hemophilia therapy. 19. A method of treating stroke in a patient by inhibiting factor XIa, comprising administering to a patient in need thereof an effective amount of a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt or composition thereof according to any one of claims 9, 10. 20. A method of reducing the risk of stroke in a patient who has suffered an ischemic attack, comprising administering to the patient in need thereof an effective amount of a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt or composition thereof according to any one of claims 9, 10. 21. A method of treating acute coronary syndrome by inhibiting factor XIa in a patient, comprising administering to a patient in need thereof a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof or a composition according to any one of claims 9, 10. 22. A method of reducing the risk of acute coronary syndrome by inhibiting factor XIa in a patient, comprising administering to a patient in need thereof a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof or a composition according to any one of claims 9, 10. 23. A method of treating a thromboembolic disorder by inhibiting factor XIa in a patient, comprising administering to a patient in need thereof a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof or a composition according to any one of claims 9, 10. 24. A method of reducing the risk of a thromboembolic disorder by inhibiting factor XIa in a patient, comprising administering to a patient in need thereof a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof or a composition according to any one of claims 9, 10. 25. The method of claim 23 or 24, wherein the thromboembolic disorder is selected from the group consisting of arterial cardiovascular thromboembolic event, arterial thrombosis, venous cardiovascular thromboembolic event, unstable angina, acute coronary syndrome, primary myocardial infarction, recurrent myocardial infarction, ischemia, stroke , atherosclerosis, peripheral arterial occlusive disease, venous thromboembolism, venous thrombosis, deep vein thrombosis, thrombophlebitis, arterial embolism, coronary artery thrombosis, cerebral artery thrombosis, cerebral embolism, renal embolism and pulmonary embolism. 26. A method of treating atrial fibrillation by inhibiting factor XIa in a patient, comprising administering to a patient in need thereof a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof or a composition according to any one of claims 9, 10. 27. A method of reducing the risk of atrial fibrillation by inhibiting factor XIa in a patient, comprising administering to a patient in need thereof an effective amount of a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt or composition thereof according to any one of claims 9, 10. 28. The method according to any one of claims 11 to 27, wherein the compound is administered orally or parenterally.