RU2789127C2 - P14kiii beta inhibitors - Google Patents

Abstract

FIELD: medicine; therapy.

SUBSTANCE: group of inventions relates to the field of medicine, namely to therapy; it is intended for the treatment of a rhinovirus infection or a disorder caused or exacerbated with a rhinovirus infection. A compound of (S)-N-(1-hydroxypropane-2-yl)-5-(5-(2-hydroxypropane-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazole-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidine-3-yl)-N,2-dimethylbenzamide is presented. In another embodiment, a pharmaceutical composition containing this compound is provided. Methods for the treatment of a rhinovirus infection or treatment of a disorder caused or exacerbated with a rhinovirus infection, including administration of the specified compound, are also provided.

EFFECT: group of inventions provides a new compound of (S)-N-(1-hydroxypropane-2-yl)-5-(5-(2-hydroxypropane-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazole-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidine-3-yl)-N,2-dimethylbenzamide, as well as a pharmaceutical composition containing this compound, which is a strong PI4KIIIβ inhibitor and is used for the treatment of a rhinovirus infection and disorders caused or exacerbated with such a virus infection.

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Description

FIELD OF TECHNOLOGY

The present invention relates to compounds that are inhibitors of kinase activity, pharmaceutical compositions containing these compounds, and variants of their use in the treatment and prevention of viral infections and disorders caused or aggravated by such a viral infection.

BACKGROUND OF THE INVENTION

The genus Enterovirus (enteroviruses) of the Picornaviridae family of single-stranded viruses containing single-stranded positive-sense RNA includes a number of human pathogens that can cause very severe disease. Examples include poliovirus, Coxsackievirus B (aseptic meningitis, myocarditis, pancrectitis, and non-specific fibrillary disease), enterovirus-A71 (aseptic meningitis, encephalitis, and polio-like paralysis), enterovirus-D68 (acute flaccid myelitis), and parechovirus (myocarditis and encephalitis). Enteroviral infections (variant A24 of the Coxsackie virus and type 70 of the enterovirus) are the cause of most cases of acute hemorrhagic conjunctivitis (AHC, ANS). Numerous epidemics and three pandemics of OHC have occurred since 1969 (Yin-Murphy et al., British Journal of Ophthalmology, 1986, 70, 869; Nilsson et al., Journal of Virology, 2008 82, 3061). However, most people with enterovirus infection develop much less serious illnesses. The common cold is one of the most common human diseases and is most often associated with another type of enterovirus, human rhinovirus (HRV), which causes 30-50% of colds.

et al., Journal of Clinical Microbiology, 1997, 35, 1791 и Clinical Infectious Diseases, 2001 33, 909) и у 29% пациентов, проходящих функциональную эндоскопическую синусохирургию по поводу хронического риносинусита (Abshirini et al., Jundishapur Journal of Microbiology, 2015, 8, e20068).The optimum temperature for HRV replication is 33-35°C, which promotes upper respiratory tract infection (URTI) and disease, which is most often mild and resolves without medical intervention. However, URTI can have complications, and HRV has been detected in the middle ear in ~40% of children under age 7 with exudative otitis media, including chronic cases (Papadopoulus et al., Pediatric Allergy and Immunology, 2006: 17: 514), in sputum in 26% of 291 patients with acute bronchitis (Park et al. Plos One, 2016, 11, e0165553), in aspirates and scrapings from the maxillary sinus in 15 of 34 patients with acute sinusitis,

et al., Journal of Clinical Microbiology, 1997, 35, 1791 and Clinical Infectious Diseases, 2001 33, 909) and in 29% of patients undergoing functional endoscopic sinus surgery for chronic rhinosinusitis (Abshirini et al., Jundishapur Journal of Microbiology, 2015 , 8, e20068).

Large and medium airway temperatures in the lungs must also allow HRV replication (McFadden ER Jr, et al., J. Appl. Physiol., 1985, 58, 564), and infection can cause very severe disease in some patient populations. In children under the age of 5, rhinovirus infection often results in hospitalization (4.8 cases/1000 children: Miller et al., Journal of Infectious Diseases, 2007; 195,773), with disease severity similar to that associated with respiratory infection. syncytial virus (RSV) (McMillan et al., Pediatric Infectious Disease Journal, 1993, 12, 321), and often leads to bronchiolitis and pneumonia (Kellner et al., Acta Pediatrica Scandinavica, 1989, 78, 390; McMillan et al. , Pediatric Infectious Disease Journal, 1993, 12, 321; El-Sahly et al., Clinical infectious Diseases, 2000, 31, 96; Jartti and Korppi, Pediatric Allergy and Immunology, 2011, 22, 350). In a study by Asner et al. (Influenza and Other espiratory Viruses, 2014, 8, 436), most children with human rhinovirus/enterovirus infection had underlying immunosuppression or comorbid cardiorespiratory disease; and it is well known that the consequences of human rhinovirus infection can be particularly severe for patients with these conditions (Anzueto et al., Chest, 2003, 123.1664; Rotbart, Antivir. Res. 2002, 53, 83). For example, 7 out of 22 rhinovirus-infected myelosuppressive transplant recipients developed pneumonia resulting in death (Ghosh et al., Clinical infectious Diseases, 1999, 29, 528).

Enteroviral infections are also often associated with skin rashes (pemphigus of the mouth and extremities, Coxsackievirus-associated eczema, and other atypical rashes: Hubisch et al., Pediatric Infectious Disease Journal, 2014, 33, e92; Korman et al., Journal of the American Academy of Dermatology, 2017, 76, 538; Drago et al., Future Microbiology, 2017 12, 171).

HRV is the virus most commonly associated with exacerbations of asthma (approximately 25% of exacerbations in adults and 50% in children, Nicholson et al., BMJ, 1993, 307.982; Johnston et al., BMJ., 199, 310.1225) and chronic obstructive pulmonary disease (COPD: 20-26% Seemungal et al., Am. J. Respir. Crit. Care Med., 2001, 164.1618; Papi et al., Am. J. Respir. Crit. Care Med. , 2006, 173. 1114), and in both cases, experimental infection with rhinovirus has been shown to exacerbate (aggravate) the disease (Zambrano et al., J Allergy Clin Immunol., 2003, 111, 1008; Mallia et al., Am J. Respir. Crit. Care Med., 2011, 183.734). Rhinovirus infections are also frequently associated with exacerbations of bronchiectasis (16–25%: Kapur et al. Arch Dis Child 2014, 99, 749; Gao et al Chest 2015, 147, 1635) and cystic fibrosis (CF, MB) (Etherington, J. Cystic Fibrosis 2014, 13, 49; Flight et al Thorax, 2014, 69.247). In COPD, MB, and bronchiectasis, exacerbations are more severe when they are associated with viral infection (Papi et al. Am. J. Respir. Crit. Care Med., 2006, 173.1114; Etherington, J. Cystic Fibrosis, 2014, 13, 49; Kapur et al., Arch Dis Child, 2014, 99, 749), and in each case, exacerbations contribute to disease progression with reduced survival (Liou et al., Am J Epidemiol., 2001, 153, 345; Soler- Cataluna et al., Thorax, 2005, 60.925; Roberts et al., Intern Med J., 2012 42, 129). Most rhinovirus-induced exacerbations of COPD are followed by a subsequent secondary bacterial infection (Mallia et al., Am. J. Respir. Crit. Care Med., 2012, 186, 1117; George et al., Eur Respir J., 2014;. 87). In addition, human rhinovirus infection is one of the factors that may direct the development of the asthmatic immune system phenotype in the infant (D.J. Jackson et al., Am. J. Respir. Crit. Care Med., 2008, 178, 667).

The socioeconomic impact of human rhinovirus is enormous, and treatment often includes the unnecessary use of antibiotics. The common cold is estimated to be responsible for at least twenty-five million absenteeisms from the workplace and about the same number of absenteeisms from school each year in the US (Rotbart, Antivir. Res., 2002 53, 83). The direct and indirect cost of the common cold and its complications in asthmatics alone in the United States is estimated at forty billion dollars annually (A. M. Fendrick et al., Arch. Intern. Med., 2003, 163, 487.

There are three types of HRV (A, B and C), including more than 150 genotypes. HRV-A and -C are the most frequently detected, and the latter seems to represent a more pathogenic group, at least in the pediatric population with asthma (Piralla et al., Journal of Clinical Virology, 2009, 45, 311; Bizzintino et al., Eur. Respir. J., 2011, 37, 1037). Rhinoviruses are also divided into 3 broad groups based on the cellular receptor that mediates their entry into the cell. The main group of human rhinoviruses (approximately 90% of the HRV-A and B verotypes) enter the cell by interacting with the human intercellular adhesion molecule (ICAM-1). The remaining ~10% of HRV-A and B viruses are a less numerous group and use the low-density lipoprotein receptor to enter the cell. More recently discovered HRV-C species bind to cadherin-related family member 3 (CDHR3) to effect entry. Human rhinoviruses enter the cell by triggering receptor-mediated endocytosis, and the disassembly of the envelope occurs in endosomes. Differences between serotypes not only prevent the development of cross-immunity in the body, but also greatly complicate the development of vaccines and other virus-specific methods of prevention and treatment.

The human rhinovirus (HRV) RNA genome without proteins (~8 kb) is surrounded by a capsid consisting of sixty copies of each of the four structural proteins, designated VP1 - VP4, in an icosahedoric configuration, forming a viral particle with a diameter of ~30 nm. HRV replication requires an RNA-dependent RNA polymerase that is dependent on viral RNA, as well as a number of viral and host cell accessory proteins. The HRV genome is translated as a single polyprotein, which, after translation, is cleaved by virally encoded proteases into three proteins, which in turn are cleaved into at least eleven proteins. Replication of the viral genome can begin as soon as one hour after infection, and as little as four hours after the entry of the virus into cells, almost one million fully assembled cell particles can be released, followed by cell death.

There is currently no approved human drug that will cure HRV infection. Several attempts to target HRV viruses directly have shown some positive results. 4-[2-[1-(6-Methyl-3-pyridazinyl)-4-piperidinyl]-ethoxy]-benzoate, also known as "pyrodavir" is able to act as a capsid-binding inhibitor, but problems with solubility, endogenous degradation, and cost limited its use against HRV. Pleconaril (PIKOVIR) has been shown to be effective in inhibiting replication but has not been approved by the US Food and Drug Administration (FDA) citing significant concerns about its safety. Rupintrivir, an inhibitor of the 3C viral protease, was effective in human studies with experimental HRV infection, but proved ineffective against natural HRV infections (Bauer et al., Current Opinion in Virology, 2017, 24, 1). It has been suggested that certain imidazopyrazines may be effective antivirals against HRV and other viruses; the mechanism of action is unclear, although it is believed not to be related to cyclin-dependent kinases (US Patent Publication 2011/0166147, Macleod et al.).

In light of the above, there remains a need for new therapeutic agents against human rhinoviruses and enteroviruses.

Despite other differences, genome replication of positive-strand RNA viruses depends on a single fundamental step in RNA-dependent RNA synthesis. This step is essential to the life cycle of the virus, and these viruses are known to require many host proteins to start and maintain RNA-dependent RNA polymerase activity. Without interaction with host factors, viruses would not be able to replicate/survive. Therefore, a possible therapeutic intervention to suppress viral infections of this class is to block the interaction of the virus with the host, especially in relation to the inhibition of viral genome replication. Manipulation of host factors that are essential to the virus but not essential to the host can achieve significant inhibition of viral spread. In addition, excess host factors may represent promising targets for intervention. This is especially true when large classes of viruses have evolved the ability to interact with only one of a number of redundant host factors. One set of host proteins that are considered potential targets for the inhibition of viral replication are phosphatidylinositol 4-kinases.

Phosphatidylinositol 4-kinases (PI4Ks) are involved in several cellular activities, including membrane fusion, vesicular transport, and cell signaling, by catalyzing the phosphatidylinositol 4-kinase to form phosphatidylinositol-4 phosphate (PI4P). There are several known PI4K isoforms that differ in several characteristics, including sequence, size, tissue and cellular localization, and function.

One type of PI4K, phosphatidylinositol (type III-4-kinase, beta polypeptide (PI4KIIIβ) (also known in the literature as phosphatidylinositol 4-kinase(III)β, Ptdlns 4-kinase(III)β, PI4Kβ, Pi4kcb and PI4K92) is important for the control of local populations of PI4P, primarily in the Golgi apparatus, where it is required to maintain the structural integrity of the organelle. This enzyme has also been found in the nucleus. Recent studies have suggested that PI4KIIIβ is involved in the replication of the genome of several RNA viruses, including human rhinovirus, enteroviruses 68 and 71, poliovirus, coxsackievirus, hepatitis C virus, bovine cobuvirus, Aichivirus, rabies virus, and others (see, for example, van der Schaar et al., Antimicrobial Agents and Chemotherapy, 57, 4971; Roulin et al., Cell Host & Microbe, 2014, 16, 677; Mello et al., Antimicrobial Agents and Chemotherapy 2014, 58, 1546; Jun Sasaki et al., EMBO J. 2011, 31 754 Hsu et al., Cell 2010, 141. 79 9; Borawski, J. Virology 2009, 83, 10058; Altan-Bonnet et al., TIBS 2012, 37, 293). In addition, the catalytic activity of PI4KB has been shown to be required for styloid-mediated cell entry of the SARS coronavirus, the virus that causes severe acute respiratory syndrome (Yang et al., J. Biol. Chem., 2012, 287, 8457). SARS is the cause of an epidemic that originated in southern China, which accounted for 8448 cases and 774 deaths in 37 countries from November 2002 to July 2003. The macroeconomic impact of this outbreak was estimated at US$30 to US$100 billion (Smith, Social Science & Medicine , 2006, 63, 3113). It is generally accepted that inhibition of PI4KB can significantly reduce viral replication in many RNA viruses and positive-strand RNA viruses in particular, making PI4KB a potential target for the development of broad-spectrum antiviral agents. Additionally, PI4 kinases are involved in bacterial entry and replication, and it has been suggested that PI4KB is involved in Legionella pneumophila infection through the role of PI4P in anchoring bacterial proteins to legionella-containing intracellular vacuoles (Clayton et al. Pu Progress in Lipid Research 2013, 52 294). Accordingly, a PI4KB inhibitor may be effective against acute lung injury or acute respiratory distress syndrome associated with legionella infections and possibly as a treatment for other bacterial infections.

E.R. Keaney et al. (Bioorg. Med. Chem. Lett., 24 (2014) 3714-3718) describe 2-alkyloxazole derivatives as PI4KIIIβ inhibitors for the potential treatment of hepatitis C virus infections.

I. Medrova et al. (J. Med. Chem., 2017, 60(1), 100-118) describe a series of imidazo[1,2-b]pyridazine derivatives as PI4KIIIβ inhibitors for possible treatment of viral infections.

J.V. Shotwell, in a slide presentation titled "Chemical Optimization of Novel Inhibitor Classes for PI4KIIIβ: A Critical Host Factor for Enterovirus Replication" submitted to "The 27th International Conference on Antiviral Research" (held in Raleigh, MA) North Carolina, USA) and presented on May 12, 2014, described a number of compounds exhibiting activity at the PI4KB receptor. The slides were primarily related to oral administration and include a slide titled "Existing Chemotyopes were optimized for IN Delivery", which showed the effect on the lungs of intranasal administration of Compound GSK3180404A and Compound GSK3159043A in rat models. This slide shows the significant accumulation of the compound in tissues for both compounds. The next slide, titled "Nasal Epithelian Findings Observed following IN dose", contains a series of histological images demonstrating ulceration in the nasal cavity in rats and bronchial epithelial hyperplasia in the lungs of rats following intranasal administration of GSK3159043.

There is a need for compounds that are potent inhibitors of PI4KIIIβ. There is also a need for compounds that can also be used as selective PI4KIIIβ inhibitors.

In particular, there is a need for compounds that are potent PI4KIIIβ inhibitors that do not accumulate to a significant extent in body tissues, such as lung tissues, especially when administered via the inhalation or intranasal routes. Such compounds are useful in the treatment of viral infections and disorders caused or exacerbated by such viral infection, in particular human rhinovirus (HRV) infections.

BRIEF DESCRIPTION OF THE INVENTION

The present invention relates to compounds of Formula (I) or their pharmaceutically acceptable salts,

where R ¹ , R ² , R ³ , R ⁴ , R ^4b , R ^4c , R ⁵ , W, X, Y and Z are defined herein.

Compounds of Formula (I) have been shown to be selective inhibitors of PI4KIIIβ and useful in the treatment or prevention of viral infections and disorders caused or aggravated by such viral infection. Disorders caused or exacerbated by viral infections include, in particular, COPD, asthma, cystic fibrosis, bronchiectasis, and congestive heart failure. Additionally, disorders caused or exacerbated by rhinovirus infections include bronchiolitis, otitis media, sinusitis, and acute bronchitis. Also, rhinovirus infections can lead to secondary bacterial infections in children, the elderly, and people with immunosuppression. Such secondary bacterial infections can cause pneumonia.

Accordingly, the present invention further relates to methods for treating or preventing viral infections and disorders caused or aggravated by such viral infection, said methods comprising administering to a patient in need thereof a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

The present invention also relates to pharmaceutical compositions containing a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients.

The present invention also relates to compounds of Formula (I) or their pharmaceutically acceptable salts for use in therapy

The present invention also relates to the use of the compounds of Formula (I) or their pharmaceutically acceptable salts in the manufacture of a medicament for the treatment or prevention of viral infections and disorders caused or aggravated by such viral infection.

DETAILED DESCRIPTION OF THE INVENTION

According to a first aspect of the present invention, there is provided a compound of general Formula (I), or a pharmaceutically acceptable salt thereof,

Where

W is C, X is C, Y is N, and Z is C;

W is C, X is N, Y is C, and Z is C;

W is C, X is N, Y is C, and Z is N;

W is N, X is C, Y is C, and Z is N; or

W is N, X is C, Y is C, and Z is C;

R ¹ is C _1-4 alkoxy, -C(=O)N(R ^1a R ^1b ), -S(=O) ₂ -N(R ^1a R ^1b ), -S(=O) ₂ -R ^1c or -S(=O)-R ^1c , and

R ^1a is C _1-3 alkyl, haloC _1-3 alkyl, hydroxyC _1-3 alkyl, C _1-3 alkoxyC _1-3 alkyl, tetrahydropyranyl or tetrahydrofuranyl; R ^1b is H or C _1-3 alkyl, or R ^1a and R ^1b together with the nitrogen to which they are attached form a 4-7 membered ring, said ring containing carbon atoms in the ring and optionally one oxygen atom in ring, wherein said ring a) is optionally substituted with one or two groups selected from C _1-3 alkyl, halogen, C _1-3 alkoxy, hydroxy, hydroxyC _1-3 alkyl and oxo, which may be the same or different or b) ortho- or spiro-fused to an unsubstituted 4-6 membered cycloalkane ring or an unsubstituted 4-6 membered saturated heterocyclic ring; And

R ^1c is C _1-3 alkyl, C _1-3 alkoxy, hydroxy, hydroxyC _1-3 alkyl, or C _1-3 alkoxyC _1-3 alkyl;

R ² is H, C _1-3 alkyl, halo, or —OR ^2a , wherein R ^2a is H or an unsubstituted linear C _1-3 apkyl chain, with one or two carbon atoms in the chain optionally replaced by oxygen atoms;

R ³ is H or halogen;

and while either

i) R ^4a is H, C _1-3 alkyl or halo, R ^4b is C _1-3 alkyl, cyclopropyl or hydroxyC _1-2 alkyl; or R ^4a and R ^4b together with the carbon atom to which they are attached form a 3-6 membered saturated ring containing ring carbon atoms and optionally a ring oxygen atom, said ring being optionally substituted with one C _1-3 alkyl group, or one hydroxyC _1-2 alkyl group; and R ^4c is OH, hydroxymethyl or hydroxyethyl;

ii) R ^4a H, C _1-3 alkyl, halo or OH; R ^4b is H, C _1-3 alkyl or halo, R ^4c is an unsubstituted ring selected from the list consisting of oxetanyl, tetrahydrofuranyl and tetrahydropyranyl; or

iii) R ^4a is H and R ^4b and R ^4c together with the carbon atom to which they are attached form an unsubstituted ring selected from the list consisting of oxetane, tetrahydrofuran or tetrahydropyran; And

R ⁵ is

a) imidazol-2-yl optionally substituted with a C _1-3 alkyl group in position 1 and optionally substituted with a methyl group in position 5; or

b) pyrazol-1-yl optionally substituted with a C _1-3 alkyl group at position 5 and optionally substituted with a methyl group at position 4.

In one embodiment, W is C, X is N,

Z is C and Y is C.

In yet another embodiment, R ^4c is OH.

In one embodiment, R ² is H, C _1-3 alkyl, chlorine, or —OR ^2a , where R ^2a is H or an unsubstituted linear C _1-3 alkyl chain, with one or two carbon atoms in the chain optionally replaced by atoms oxygen; and R ³ is H or fluoro;

In another embodiment i) R ^4a is H, C _1-3 alkyl or fluorine; R ^4b is C _1-3 alkyl, cyclopropyl or hydroxyC _1-2 alkyl; or R ^4a and R ^4b together with the carbon atom to which they are attached form a 3-6 membered saturated ring containing ring carbon atoms and optionally a ring oxygen atom, said ring being optionally substituted with one C _1-3 alkyl group, or one hydroxyC _1-2 alkyl group; and R ^4c is OH, hydroxymethyl or hydroxyethyl;

ii) R ^4a H, C _1-3 alkyl, fluoro or OH; R ^4b is H, C _1-3 alkyl or fluoro; R ^4c is an unsubstituted ring selected from the list consisting of oxetanyl, tetrahydrofuranyl and tetrahydropyranyl; or

iii) R ^4a is H and R ^4b and R ^4c together with the carbon atom to which they are attached form an unsubstituted ring selected from the list consisting of oxetane, tetrahydrofuran or tetrahydropyran;

In one embodiment, the present invention provides compounds of Formula (Ia) or pharmaceutically acceptable salts thereof:

Where

X is N or C, Y is N or C, and Z is N or C; wherein X and Y cannot both be N or both be C; and wherein if Z is N, X is N, and Y is C;

R ¹ is C _1-4 alkoxy, -C(=O)N(R ^1a R ^1b ), -S(=O) ₂ -N(R ^1a R ^1b ), -S(=O) ₂ -R ^1c or -S(=O)-R ^1c , where

R ^1a is C _1-3 alkyl, haloC _1-3 alkyl, hydroxyC _1-3 alkyl or C _1-3 alkoxyC _1-3 alkyl; R ^1b is H or C _1-3 alkyl, or R ^1a and R ^1b together with the nitrogen to which they are attached form a 4-7 membered ring, said ring containing carbon atoms in the ring and optionally one oxygen atom in ring, wherein said ring is a) optionally substituted with one or two groups selected from C _1-3 alkyl, halogen, C _1-3 apkoxy, hydroxy and oxo, which may be the same or different, or b) ortho- or spiro-fused with an unsubstituted 4-6 membered cycloalkane ring or an unsubstituted 4-6 membered saturated heterocyclic ring; And

R ^1c is C _1-3 alkyl, C _1-3 alkoxy, hydroxy, hydroxyC _1-3 alkyl, or C _1-3 alkoxyC _1-3 alkyl;

R ² is H, C _1-3 alkyl, chlorine, or —OR ^2a where R ^2a is n or an unsubstituted linear C _1-3 alkyl chain, with one or two carbon atoms in the chain optionally replaced by oxygen atoms;

R ³ is H or fluoro;

R ^4a is H or methyl;

R ^4b is C _1-3 alkyl or hydroxyC _1-2 alkyl; And

R ⁵ is

a) imidazol-2-yl optionally substituted with a C _1-3 alkyl group in position 1 and optionally substituted with a methyl group in position 5; or

b) pyrazol-1-yl optionally substituted with a C _1-3 alkyl group at position 5 and optionally substituted with a methyl group at position 4.

In one embodiment, X is N, Z is C, and Y is C.

In one embodiment, the compound is a compound of Formula (Ib):

In another embodiment, the compound is a compound of Formula (Ic):

In one embodiment, is -C(=O)N(R ^1a R ^1b ) or -S(=O) ₂ -R ^1c . In another embodiment, R ¹ is -C(=O)N(R ^1a R ^1b ).

In one embodiment, R ^1a is hydroxyC _1-3 alkyl or tetrahydropyranyl. In one embodiment, R ^1a is hydroxyC _1-3 alkyl. In yet another embodiment, R ^1a is 3-hydroxy-1-propyl, 2-hydroxy-1-ethyl, 3-hydroxy-2-propyl, or 4-tetrahydropyranyl.

In one embodiment, R ^1b is C _1-3 alkyl. In another embodiment, R ^1b is methyl or ethyl.

In yet another embodiment, R ^1a is hydroxyC _1-3 alkyl and R ^1b is C _1-3 alkyl. In yet another embodiment, R ^1a is 3-hydroxy-1-propyl and R ^1b is C _1-3 alkyl. In another embodiment, R ^1a is 3-hydroxy-2-propyl and R ^1b is C _1-3 alkyl. In another embodiment, R ^1a is 3-hydroxy-1-propyl and R ^1b is methyl. In another embodiment, R ^1a is 3-hydroxy-2-propyl and R ^1b is methyl. In another embodiment, R ^1a is 2-hydroxy-1-ethyl and R ^1b is ethyl. In another embodiment, R ^1a is 3-hydroxy-2-propyl and R ^1b is methyl. In another embodiment, R ^1a is 4-tetrahydropyranyl and R ^1b is methyl.

In one embodiment, R ^1a and R ^1b , together with the nitrogen to which they are attached, form a 4- to 7-membered saturated ring, said ring containing ring carbon atoms and optionally one ring oxygen atom, said ring a ) is optionally substituted with one or two groups selected from C _1-3 alkyl, halogen, C _1-3 alkoxy, hydroxy, hydroxyC _1-3 alkyl and oxo, which may be the same or different, or b) ortho- or spiro-fused with an unsubstituted 4-6 membered cycloalkane ring; or an unsubstituted 4-6 membered saturated heterocyclic ring.

In one embodiment, R ^1a and R ^1b , together with the nitrogen to which they are attached, form an optionally substituted pyrrolidine ring. In yet another embodiment, the pyrrolidine ring is substituted with C _1-3 alkyl, hydroxy, or hydroxyC _1-3 alkyl.

In one embodiment, R ^1c is hydroxyC _1-3 alkyl. In another embodiment, R ^1c is 2-hydroxy-1-ethyl.

In one embodiment, R ² is C _1-3 alkyl, chlorine or —OR ^2a . In yet another embodiment, R ² is C _1-3 alkyl, chlorine, or methoxy.

In one embodiment, R ³ is H or fluoro; In another embodiment, R ³ is H. In one embodiment, R ^4a is methyl.

In one embodiment, R ^4b is C _1-3 alkyl. In another embodiment, R ^4b is methyl or ethyl. In another embodiment, R ^4b is methyl.

In one embodiment, R ^4a is methyl and R ^4b is methyl.

In one embodiment, R ^4a is C _1-3 alkyl, R ^4b is C _1-3 alkyl, and R ^4c is OH. In another embodiment, R ^4a is methyl, R ^4b is methyl, and R ^4c is OH.

In one embodiment, R ⁵ is imidazol-2-yl optionally substituted with a C _1-3 alkyl group at position 1 and optionally substituted with a methyl group at position 5. In another embodiment, R ⁵ is 1-methyl-1H- imidazol-2-yl.

In one embodiment, said compound is a compound according to formula (I) and:

W is C, X is N, Z is C, and Y is C;

R ¹ is -C(=O)N(R ^1a R ^1b ) or -S(=O) ₂ -R ^1c where R ^1a is hydroxyC _1-3 alkyl and R ^1b is C _1-3 alkyl ; or R ^1a and R ^1b , together with the nitrogen to which they are attached, form an optionally substituted pyrrolidine ring; and wherein R ^1c is hydroxyC _1-3 alkyl;

R ² is C _1-3 alkyl, chlorine or —OR ^2a ;

R ³ is H;

R ^4a is methyl;

R ^4b is C _1-3 alkyl;

R ^4c is OH; And

R ⁵ is imidazol-2-yl optionally substituted with a C _1-3 alkyl group at position 1 and optionally substituted with a methyl group at position 5.

In one embodiment, said compound is a compound according to formula (I) and:

W is C, X is N, Z is C, and Y is C;

R ¹ is -C(=O)N(R ^1a R ^1b ), where R ^1a is hydroxyC _1-3 alkyl and R ^1b is C _1-3 alkyl; or R ^1a and together with the nitrogen to which they are attached form an optionally substituted pyrrolidine ring substituted with C _1-3 alkyl, hydroxy, hydroxyC _1-3 alkyl;

R ² is C _1-3 alkyl, chlorine or methoxy;

R ³ is H;

R ^4a is methyl;

R ^4b is methyl;

R ^4c is OH; And

R ⁵ is 1-methyl-1H-imidazol-2-yl.

In one embodiment, the compound of Formula (I) is selected from the group consisting of the following compounds:

2-chloro-N-ethyl-N-(2-hydroxyethyl)-5-(5-(1-hydroxyethyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl) benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)benzamide (compound 15);

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-N-(3-hydroxypropyl)-N,2-dimethylbenzamide (compound 17);

N-ethyl-N-(2-hydroxyethyl)-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl) benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-2-methoxybenzamide (compound 19);

(S)-N-(1-hydroxypropan-2-yl)-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazole- 2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-2-methoxy-N-methylbenzamide (compound 20);

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-2-methoxy-N-methyl-N-(tetrahydro-2H-pyran-4-yl)benzamide (compound 21);

(S)-N-(1-hydroxypropan-2-yl)-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazole- 2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-[N,2-dimethylbenzamide (compound 22);

(R)-(2-(hydroxymethyl)pyrrolidin-1-yl)(5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazole -2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-2-methylphenyl)methanone (compound 25);

(S)-(5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1 ,5-a]pyrimidin-3-yl)-2-methylphenyl)(3-hydroxypyrrolidin-1-yl)methanone (compound 29);

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-N-(3-hydroxypropyl)-2-methoxy-N-methylbenzamide (compound 32); And

(R)-(2-(hydroxymethyl)pyrrolidin-1-yl)(5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazole -2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-2-methoxyphenyl)methanone (compound 36);

N-ethyl-N-(2-hydroxyethyl)-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl) benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-2-methylbenzamide (compound 38);

N-(2-hydroxyethyl)-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino) pyrazolo[1,5-a]pyrimidin-3-yl)-N,2-dimethylbenzamide (compound 43);

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-N,2-dimethyl-N-(tetrahydrofuran-3-yl)benzamide (compound 59);

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-N-(2-hydroxypropyl)-N,2-dimethylbenzamide, isomer 1 (compound 62);

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-а] pyrimidin-3-yl)-N-(2-hydroxypropyl)-N,2-dimethylbenzamide, isomer 2 (compound 63);

2-Chloro-N-(2-hydroxyethyl)-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl) benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-N-methylbenzamide (compound 37);

N-((S)-1-hydroxypropan-2-yl)-5-(5-(1-hydroxypropyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl) benzyl)amino)pyrazolo[1,5-a]pyrimidin3-yl)-N,2-dimethylbenzamide, isomer 1 (compound 80);

N-((S)-1-hydroxypropan-2-yl)-5-(5-(1-hydroxypropyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl) benzyl)amino)pyrazolo[1,5-a]pyrimidin3-yl)-N,2-dimethylbenzamide, isomer 2 (compound 81);

5-(5-(1-hydroxybuten-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-N-(3-hydroxypropyl)-N,2-dimethylbenzamide, isomer 1 (compound 66); And

5-(5-(1-hydroxybuten-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-N-(3-hydroxypropyl)-N,2-dimethylbenzamide, isomer 2 (compound 67);

or pharmaceutically acceptable salts of any of the above compounds.

In one embodiment, the compound of Formula (I) is 5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl) benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-N-(3-hydroxypropyl)-N,2-dimethylbenzamide (compound 17)

or a pharmaceutically acceptable salt thereof.

In one embodiment, the compound of Formula (I) is N-ethyl-N-(2-hydroxyethyl)-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-( 1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-2-methoxybenzamide (compound 19)

or a pharmaceutically acceptable salt thereof.

In one embodiment, the compound of Formula (I) is 5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl) benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-2-methoxy-N-methyl-N-(tetrahydro-2H-pyran-4-yl)benzamide (compound 21)

or a pharmaceutically acceptable salt thereof.

In one embodiment, the compound of Formula (I) is (S)-N-(1-hydroxypropan-2-yl)-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-( (3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-N,2-dimethylbenzamide (compound 22)

or a pharmaceutically acceptable salt thereof.

Terms and Definitions

Alkyl is a monovalent radical formed by the removal of a hydrogen atom from an acyclic alkane. For example, a C _1-4 alkyl is alkyl having 1 to 4 carbon atoms. The alkyl may be linear or branched. Examples of C _1-4 alkyl are methyl, ethyl, n-propyl, n-butyl, isopropyl, isobutyl, sec-butyl tert-butyl.

Alkoxy is a group of Formula "-OR" where R is alkyl (as defined above). For example, C _1-4 alkoxy is an alkoxy consisting of 1 to 4 carbon atoms. Examples of C _1-4 alkoxy are methoxy, ethoxy, n-propoxy, n-butoxy, iso-propoxy, iso-butoxy, sec-butoxy and t-butoxy.

Halo (halogen) refers to the hapogene radical, i.e., fluorine-, chlorine-, bromine- or iodine-.

Haloalkyl is alkyl (as defined above) substituted with one or more halogens ("halo" as defined above), the halogens being the same or different. For example, haloC _1-3 alkyl is a haloalkyl of 1 to 3 carbon atoms. Examples of haloC _1-3 alkyl are monofluoromethyl, difluoromethyl, trifluoromethyl and 1-chloro-2-fluoroethyl.

Hydroxyalkyl is alkyl (as defined above) substituted with one or more hydroxy substituents. For example, hydroxyC ₃ alkyl has the formula -(CH ₂ ) ₃ OH (where "-" indicates which atom is attached to the compound of Formula (I)).

Alkoxyalkyl is alkyl (as defined above) substituted with one or more alkoxy substituents. For example, C ₃ alkoxyC ₂ alkyl has the formula -(CH ₂ ) ₂ O(CH ₂ ) ₂ CH ₃ (where "-" indicates which atom is attached to the compound of Formula (I)).

Oxo is a divalent radical of the formula =O.

The 4-6 membered saturated heterocyclic ring is monocyclic and is composed of ring carbon atoms and ring heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. In one embodiment, the heterocyclic ring contains 1 or 2 heteroatoms in the ring. Examples are pyrrolidine, dioxolane, imidazolidine, pyrazolidine, piperidine, dioxane, morpholine, dithian, thiomorpholine and piperazine.

The 4-6 membered cycloalkane ring does not contain a single heteroatom in the ring and is saturated and monocyclic. Examples are cyclobutane, cyclopentane and cyclohexane.

An "ortho-conjugated ring system" contains two rings sharing only two atoms in common and one bond in common, for example:

A "spiro-conjugated ring system" contains two rings connected by a single carbon atom, for example,

"Substituted" in relation to a group indicates that the hydrogen atom associated with a group member atom has been replaced. It is understood that the term "substituted" implies that such a substitution corresponds to the allowable valency of the atom containing the substituent and the substituent and that the substitution yields a stable compound includes (i.e., a compound that does not undergo spontaneous transformations such as rearrangement, cyclization, or elimination ). In some embodiments, one atom may be substituted with more than one substituent, provided that such substitution corresponds to the allowable valency of that atom. Suitable substituents are defined herein for each substituted or possibly substituted group.

"Pharmaceutical acceptable" refers to compounds, materials, formulations and dosage forms which, within the reasonable medical judgment, are included within the scope of the present invention, are suitable for use in contact with human tissues without undue toxicity, irritation, or other problems or complications, and when reasonable risk/benefit ratio.

Throughout the description and the following claims, unless the context otherwise requires, the word "comprise" and its variants, such as "comprises" and "comprising" will be understood to imply the inclusion of the specified number object, step or group of objects, but not exclusion of any other object, stage or group of objects or stages.

The compounds of Formula (I) and formula (Ia) and their pharmaceutically acceptable salts may exist in solid or liquid form. In the solid state, they may exist in crystalline or non-crystalline form, or as mixtures thereof. In the case of a crystalline form, the skilled artisan will appreciate that pharmaceutically acceptable solvates can be formed in which solvent molecules are incorporated into the crystal lattice during crystallization. Solvates include non-aqueous solvents such as ethanol, isopropyl alcohol, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), acetic acid, ethanolamine, and ethyl acetate, or they may include water as a solvent embedded in the crystal lattice. Solvates in which water is the solvent embedded in the crystal lattice are commonly referred to as "hydrates". Hydrates include stoichiometric hydrates as well as compositions containing varying amounts of water.

Compounds of Formula (I) and formula (Ia) and their pharmaceutically acceptable salts that exist in crystalline form, including various solvates of such compounds, may exhibit polymorphism (ie, the ability to exist in different crystal structures). These various crystalline forms are commonly known as "polymorphs". The present invention includes all such polymorphs. The polymorphs have the same chemical composition but differ in packaging, geometric arrangement, and other properties that describe the solid crystalline state. Accordingly, polymorphs may have different physical properties such as shape, density, hardness, deformability, stability and dissolution properties, IR spectrum and X-ray powder diffraction pattern, which can be used for identification. It is understood that various polymorphs can be obtained, for example, by changing or adjusting the reaction conditions or reagents used in the preparation of the compound. For example, changes in temperature, pressure, or solvent can produce polymorphs. Additionally, one polymorph can spontaneously transform into another polymorph under certain conditions.

The invention also includes isotopically labeled compounds which are identical to the compounds of formula (I) and formula (Ia) and their pharmaceutically acceptable salts, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass numbers most common in nature. Examples of isotopes that may be included in the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen and fluorine such as ³ H, ¹¹ C, ¹⁴ C and ¹⁸ F.

Compounds corresponding to formula (I) and formula (Ia) may contain one or more centers of asymmetry (also referred to as chiral centers) and may, respectively, exist as single enantiomers, diastereoisomers or other stereoisomeric forms, or as mixtures thereof. Chiral centers such as carbon atoms may also be present on a substituent such as an alkyl group. In cases where the stereochemistry of a chiral center present in formula (I) or any other chemical structure shown here is not indicated, this structure is intended to cover any stereoisomers and all mixtures thereof. Thus, compounds corresponding to formula (I) containing one or more chiral centers can be used as racemic mixtures and racemates, enantiomerically enriched mixtures or enantiomerically pure individual stereoisomers.

Individual stereoisomers of compounds of formula (I) and formula (Ia) that contain one or more asymmetric centers can be separated by methods known to those skilled in the art. For example, such resolution may be accomplished by (1) formation of diastereomeric salts, complexes, or other derivatives; (2) by selective reaction with a stereoisomer-specific reagent, for example by enzymatic oxidation or reduction; or (3) gas-liquid or liquid chromatography in a chiral medium, for example, on a chiral carrier such as silica, with an associated chiral ligand, or in the presence of a chiral solvent. It is understood that when the desired stereoisomer is converted to another chemical compound using one of the separation procedures described above, an additional step is required to release the desired form. Alternatively, some stereoisomers can be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer to another by asymmetric conversion.

It is understood that when the desired stereoisomer is converted to another chemical compound using one of the separation procedures described above, an additional step is required to release the desired form. Alternatively, specific stereoisomers can be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts, or solvents, or by converting one enantiomer to another by asymmetric conversion.

It is to be understood that reference herein to a compound of Formula (I) and formula (Ia) or a pharmaceutically acceptable salt thereof includes a compound of Formula (I) and formula (Ia), respectively, as the free base or as a pharmaceutically acceptable salt thereof. Thus, in one embodiment, the invention relates to a compound of Formula (I). In another embodiment, the invention provides a pharmaceutically acceptable salt of a compound of Formula (I).

Pharmaceutically acceptable salts include, inter alia, those described in Berge, J. Pharm. Sci., 1977, 66, 1-19, or listed in P H Stahl, C G Wermuth, editors, Handbook of Pharmaceutical Salts; Properties, Selection and Use, Second Edition Stahl/Wermuth: Wiley-VCH/VHCA, 2011

Salts that are not pharmaceutically acceptable can be used, for example, as intermediates for the preparation of a compound of Formula (I) or formula (Ia) or a pharmaceutically acceptable salt thereof. Alternatively, non-pharmaceutically acceptable salts of Formula (I) and formula (Ia) are included herein.

Suitable pharmaceutically acceptable salts may include acid addition salts.

Such acid addition salts may be formed by reacting a compound of Formula (I) or formula (Ia) (which, for example, contains a basic amine or other basic functional group) with an appropriate acid, optionally in a suitable solvent such as an organic solvent, resulting in resulting in a salt that can be isolated by various methods, including crystallization and filtration.

Salts can be obtained in situ during the final isolation and purification of the compound of Formula (I) or formula (Ia). Where the base compound of Formula (I) or formula (Ia) is isolated as a salt, the corresponding free base form of that compound may be prepared by any suitable method known in the art, including treatment of the salt with an inorganic or organic base.

It will be appreciated that if a compound of Formula (I) or formula (Ia) contains two or more basic moieties, the salt formation stoichiometry may include 1, 2 or more equivalents of acid. Such salts may contain 1, 2 or more acid counterions, for example the dihydrochloride salt. Stoichiometric and non-stoichiometric forms of a pharmaceutically acceptable salt of a compound of Formula (I) or formula (Ia) are included within the scope of the invention, including substoichiometric salts, for example, where the counterion contains more than one acidic proton.

Typical pharmaceutically acceptable addition salts include, but are not limited to: 4-acetamidobenzoate, acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate (besylate), benzoate, bisulfate, biatrate, butyrate, calcium edetate, camphorate, camphorsulfonate (camsylate), caprate (decanoate), caproate (hexanoate), caprylate (octanoate), cinnamate, citrate, cyclamate, digluconate, 2,5-dihydroxybenzoate, disuccinate, dodecyl sulfate (estolate), edetate (ethylenediaminetetraacetate), estolate (lauryl sulfate), ethane-1, 2-disulfonate (edisylate), ethanesulfonate (esilate), formate, fumarate, galactarate (mucat), gentisate (2,5-dihydroxybenzoate), glucoheptonate (gluceptate), gluconate, glucuronate, glutamate, glutarate, glycerophosphate, glycolate, hexylresorcinate, hippurate , hydrabamine (N,N'-di(dehydroabietyl)-ethylenediamine), hydrobromide, hydrochloride, hydroiodide, hydroxynaphthoate, isobutyrate, lactate, lactobionate, laurate, malate, maleate, malonate, mandelate, methanesulfonate (mesylate), methyl sulfate, mukat, naphthalene -1 ,5-disulfonate (napadisylate), naphthalene-2-sulfonate (napsilate), nicotinate, nitrate, oleate, palmitate, p-aminobenzenesulfonate, p-aminosalicylate, pamoate (embonate), pantotheate, pectinate, persulfate, phenylacetate, phenylethylbarbiturate, phosphate, polygalactouronate, propionate, p-toluenesulfonate (tosylate), pyroglutamate, pyruvate, salicylate, sebacate, stearate, subacetate, succinate, sulfamate, sulfate, tannate, tartrate, theoclate (8-chlorotheophyllinate), thiocyanate, triethiodide, undecanoate, undecylenate, and valerate.

The compounds of Formula (I) or formula (Ia) and their salts and pharmaceutically acceptable salts include solvates (including hydrates), complexes, polymorphs, prodrugs, radiolabeled derivatives and stereoisomers of compounds of Formula (I) or formula (Ia) and their pharmaceutically acceptable salts, hereinafter referred to as "compounds according to the present invention".

Common Paths

Compounds according to the present invention can be obtained in various ways. In the following reaction schemes and beyond, unless otherwise indicated, R ¹ - R ⁵ , R ^4a , R ^4b , R ^4c , R ⁵ , W, X, Y and Z correspond to the definitions for the first aspect. Throughout the text of the application, the general formulas are indicated by Roman numerals (I), (II), (III), (IV), etc.

General way to obtain formula (I)

Compounds of formula (Ix), i.e., compounds of Formula (I) in which R ^4a is H and R ^4c is OH, can be prepared according to Reaction Scheme 1 by treating (II) with an appropriate Grignard reagent ( such as methylmagnesium bromide) in a solvent (eg THF) followed by deprotection with an appropriate acid (eg 4M HCl in 1,4-dioxane) in a solvent (eg methanol).

Scheme 1

Compounds of Formula (II) can be prepared according to Reaction Scheme 2 by treating compounds of Formula (III) with boronic ether (for example, 2-((2-methoxy-5-(4,4,5,5-tetramethyl-1,3 ,2-dioxaborolan-2-yl)phenyl)sulfonyl)ethan-1-ol) in a solvent (eg 1,4-dioxane and water) in the presence of a catalyst [eg PdCl ₂ (dppf)] and a base (eg fluoride potassium).

Scheme 2

Compounds of Formula (III) can be prepared according to Reaction Scheme 3 from compounds of Formula (IV) by treating (IV) with a suitable oxidizing agent (eg DMP) in a solvent (eg DCM).

Scheme 3

Compounds of Formula (IV) can be prepared according to Reaction Scheme 4 from compounds of Formula (V). First, the amine is protected with a suitable amine protecting group (eg t-butyl carbamate by treatment with suitable reagents eg di-t-butyl dicarbonate, DIPEA and DMAP in DCM). The ester is then reduced to the primary alcohol with a suitable reducing agent (eg sodium borohydride) in a solvent (eg ethanol).

Scheme 4

Compounds of Formula (V) can be prepared according to Reaction Scheme 5 from compounds of Formula (VI) (where L is Cl or Br) or a mixture of compounds of Formula (VI) where L is Cl and Br) by treatment with an amine (for example , (3-(1-methyl-1H-imidazol-2-yl)phenyl)methylamine) and a base (eg DIPEA) in a solvent (eg DMSO).

Scheme 5

Compounds of Formula (Ix'), i.e., compounds of Formula (I) wherein R ^4a and R ^4b are C _1-3 alkyl and R ^4c is OH, can be prepared according to Reaction Scheme 6 by treatment (VII) boronic ester (e.g. N-(3-hydroxypropyl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide ) in the presence of a catalyst [eg PdCl ₂ (dppf)] in a solvent (eg 1,4-dioxane and water) in a base (eg sodium carbonate).

Scheme 6

Compounds of Formula (VII) can be prepared from compounds of Formula (V) according to Reaction Scheme 7 by treating (V) with a Grignard reagent (eg methylmagnesium bromide) in a solvent (eg DCM). Preparation (V) is shown in Scheme 5.

Scheme 7

Compounds of Formula (VIa), i.e., compounds of Formula (VI) (see Scheme 5) wherein W is C, X is N, Y is C, Z is C, and L is Cl , can be prepared according to Reaction Scheme 8. First, compounds of Formula (VIII) can be prepared by condensing esters (such as the sodium salt of diethyloxalacetate) and compounds of Formula (IX) using an acid (such as HCl) in a solvent (such as ethanol) with heating (eg 85° C.) to give compounds of formula (VIII). Then, treatment of (VIII) with a chlorinating reagent (eg POCl ₃ ) with heating (eg at 90° C.) gives compounds of Formula (X). Treatment of compounds of Formula (X) with a source of iodine or bromine [eg N-iodosuccinimide (N-iodosuccinimide) or N-bromosuccinimide (BSI)] in a solvent (eg DCM) gives compounds of Formula (VIa).

Scheme 8

A mixture of compounds of Formula (VIb), i.e., compounds of Formula (VI) (see scheme 5), in which W is C, X is C, Y is N, Z is C, and L is Cl and Br can be obtained according to Reaction Scheme 9 in several steps from compounds (XI). First, compounds of Formula (XI) are treated with a brominating agent eg N-bromosuccinimide and sodium bicarbonate in a solvent (eg methanol) to give compounds of formula (XII). Then treatment of the compounds of Formula (XII) with 1-chloropropan-2-one at elevated temperature (eg at 90°C) gives a mixture of compounds of Formula (XIII). Iodination or bromination of a mixture of compounds of Formula (XIII), eg with N-iodosuccinimide or N-bromosuccinimide, in a solvent (eg DMF) gives a mixture of compounds of Formula (VIb).

Scheme 9

Compounds of Formula (VIc), i.e., compounds of Formula (VI) (see Scheme 5), where W is C, X is N, Y is C, Z is N, and L is Cl, may be prepared in several steps from compounds of Formula (XIV) according to Reaction Scheme 10. The compounds of Formula (XIV) may first be reacted with methyl cyanoformate to give compounds (XV). Compounds of Formula (XV) can then be cyclized using carbonyldiimidazole in a solvent (eg DMSO) or alternatively using diethyl carbonate in sodium ethoxide and ethanol to give compounds of formula (XVI). The compounds of Formula (XVI) can then be chlorinated using, for example, POCl ₃ , with heating (eg at 90° C.) to give compounds of formula (XVII). Iodination (using eg N-iodosuccinimide) or bromination (using eg N-bromosuccinimide) in a solvent (eg DMF) gives compounds of Formula (VIc).

Scheme 10

Compounds of Formula (Ix''), ie compounds of Formula (I) in which R ^4c is hydroxymethyl, can be prepared according to Reaction Scheme 11 from compounds of Formula (XVIII).

Scheme 11

Compounds of Formula (XVIIIa), i.e., compounds of Formula (XVIII) in which W is C, X is N, Y is C, and Z is C, can be prepared according to Reaction Scheme 12.

Scheme 12

Compounds of Formula (XVIIIb), i.e., compounds of Formula (XVIII) (from Scheme 11) in which W is N, X is C, and Y is C, can be prepared from compounds of Formula (XIX) in in accordance with reaction scheme 13.

Scheme 13

Alternatively, compounds of Formula (Ia) (see Scheme 1) can be prepared from compounds of Formula (V) (see Scheme 4) according to Reaction Scheme 14. Compounds of Formula (Ia) can be separated into individual enantiomers using techniques familiar to the experienced chemist, such as chiral HPLC.

Scheme 14

Compounds of Formula (Id), i.e., compounds of general formula (I) in which R ^4a and R ^4b together with the carbon atom to which they are attached form a cyclopropane ring substituted with an R group which is C _1-3 an alkyl group may be prepared according to Reaction Scheme 15 from compounds of Formula (XXa). In Reaction Scheme 15, PG is a protecting group such as t-butyloxycarbonyl and [Si] is a silicon-based protecting group for an alcohol group such as TBDMS.

Scheme 15

Compounds of Formula (Ie), i.e., compounds of Formula (I) in which R ^4a is H and R ^4d and R ^4c together with the carbon atom to which they are attached form an unsubstituted tetrahydropyran ring, can be prepared from compounds of Formula (XXI) according to Reaction Scheme 15.

Scheme 15

Compounds of Formula (XXI) can be prepared from compounds of Formula (XXII) according to the reaction scheme (16).

Scheme 16

For an experienced chemist it is clear that the compounds of Formula (I) can also be obtained by transformation in position R ¹ in the last step. In Reaction Scheme 17, compounds of Formula (If), i.e., compounds of Formula (I) in which R ¹ is -C(=O)N(R ^1a R ^1b ), can be prepared from compounds of Formula (XXIII) .

Scheme 17

It will be appreciated by the skilled chemist that compounds of Formula (I) can be converted to other compounds of Formula (I) by methods known in the art. Additionally, the intermediates described in the reaction schemes above can be converted to other intermediates and then converted using the methods described to compounds of Formula (I). It is also understood that the compounds of Formula (I) can be obtained using a different sequence of transformations described in the reaction schemes, including the appropriate steps of the introduction/removal of protection.

General Path Formula (Ia)

Compounds of Formula (Ix*), i.e., compounds of Formula (Ia) in which R4a is H, can be prepared according to Reaction Scheme 1* by treating (IIa*) with a suitable Grignard reagent (such as methylmagnesium bromide) in a solvent (eg THF) followed by deprotection with an appropriate acid (eg 4 M HCl in 1,4-dioxane) in a solvent (eg methanol).

Scheme 1*

Compounds of Formula (IIa*) can be prepared according to Reaction Scheme 2a* by treating compounds of Formula (IIIa*) with boronic ether (for example, 2-((2-methoxy-5-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-yl)phenyl)sulfonyl)ethan-1-ol) in a solvent (eg 1,4-dioxane and water) in the presence of a catalyst (eg PdCl ₂ (dppf)) and a base ( e.g. potassium fluoride).

Scheme 2a*

Compounds of Formula (III*) can be prepared according to Reaction Scheme 3* from compounds of Formula (IV*) by treating (IV*) with a suitable oxidizing agent (eg DMP) in a solvent (eg DCM).

Scheme 3*

Compounds of Formula (IV*) can be prepared according to Reaction Scheme 4* from compounds of Formula (V*). First, the amine is protected with a suitable amine protecting group (eg t-butyl carbamate by treatment with suitable reagents eg di-t-butyl dicarbonate, DIPEA and DMAP in DCM). The ester is then reduced to the primary alcohol with a suitable reducing agent (eg sodium borohydride) in a solvent (eg ethanol).

Scheme 4*

Compounds of Formula (V*) can be prepared according to Reaction Scheme 5* from compounds of Formula (VI*) (where L is Cl or Br) or a mixture of compounds of Formula (VI) where L is Cl and Br) by treatment an amine (eg (3-(1-methyl-1H-imidazol-2-yl)phenyl)methylamine) and a base (eg DIPEA) in a solvent (eg DMSO).

Scheme 5*

Compounds of Formula (Ib*), i.e., compounds of Formula (I*) wherein R ^4a and R ^4b are methyl, can be prepared according to Reaction Scheme 6 by treating (VII*) with boronic ether (e.g. N -(3-hydroxypropyl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide) in the presence of a catalyst (for example, PdCl ₂ (dppf)) in a solvent (eg 1,4-dioxane and water) with a base (eg sodium carbonate).

Scheme 6*

Compounds of Formula (VII*) can be prepared from compounds of Formula (V*) according to Reaction Scheme 7* by treating (V*) with a Grignard reagent (eg methylmagnesium bromide) in a solvent (eg DCM).

Scheme 7*

Compounds of Formula (Via*), i.e., compounds of Formula (VI*) (see Scheme 5*), where X is N, Y is C, Z is C, and L is Cl, can be obtained in accordance with the reaction scheme 8*. First, compounds of Formula (VIII*) can be prepared by condensing esters (for example, the sodium salt of diethyl oxalacetate) and compounds of Formula (IX*) using an acid (such as HCl) in a solvent (such as ethanol) under heating (for example, 85° C) to obtain a compound of formula VIII*). Then treatment of (VIII*) with a chlorinating agent (eg POCl ₃ ) with heating (eg at 90° C.) gives compounds of Formula (X*). Treatment of compounds of Formula (X*) with an iodine source (eg N-iodosuccinimide) in a solvent (eg DCM) gives compounds of Formula (Via*).

Scheme 8*

A mixture of compounds of Formula (VIb*), i.e., compounds of Formula (VI*) (see Scheme 5*), in which X is C, Y is N, Z is C, and L is Cl and Br can be obtained according to Reaction Scheme 9 in several steps from compounds (XI*). First, compounds of Formula (XI*) are treated with a brominating agent, eg N-bromosuccinimide and sodium bicarbonate in a solvent (eg methanol), resulting in compounds of formula (XII*). Then treatment of compounds of Formula (XII*) with 1-chloropropan-2-one at elevated temperature (eg 90°C) gives a mixture of compounds of Formula (XIII*). Iodination of a mixture of compounds of Formula (XIII*), eg N-iodosuccinimide, in a solvent (eg DMF) gives a mixture of compounds of Formula (VIb*).

Scheme 9*

Compounds of Formula (Vic*), i.e., compounds of Formula (VI*) (see Scheme 5*), in which X is N, Y is C, Z is N, and L is CI, can be obtained in several steps from compounds of Formula (XIV*) according to Reaction Scheme 10*. First, the compounds of Formula (XIV*) can be reacted with methyl cyanoformate to give compounds (XV*). Compounds of Formula (XV*) can then be cyclized using carbonyldiimidazole in a solvent (eg DMSO) or alternatively using diethyl carbonate in sodium ethoxide and ethanol to give compounds (XVI*). Compounds of Formula (XVI*) can then be chlorinated using, for example, POCl ₃ , with heating (eg at 90° C.) to give compounds of formula (XVII*). Iodization using, for example, N-iodosuccinimide, in a solvent (eg, DMF) gives compounds of Formula (Vic*).

Scheme 10*

It will be understood by the skilled chemist that compounds of Formula (Ia) can be converted to other compounds of Formula (Ia) by methods known in the art. Additionally, the intermediates described in the above reaction schemes can be converted to other intermediates and then can be converted, using the methods described, to compounds of Formula (Ia). It is also understood that the compounds of Formula (Ia) can be obtained using a different sequence of transformations described in the reaction schemes, including the appropriate steps of introduction/removal of protection. Methods of application

The compounds of the present invention have been shown to be potent PI4KIIIβ inhibitors. In addition, the compounds of the present invention are selective PI4KIIIβ inhibitors. The compounds of the present invention may be used in the treatment or prevention of viral infections and disorders caused or aggravated by such viral infection. In particular, diseases caused or exacerbated by viral infections include COPD, asthma, cystic fibrosis, bronchiectasis, congestive heart failure, acute respiratory distress syndrome, and acute lung injury. In addition, diseases caused or exacerbated by rhinovirus infections include bronchiolitis, otitis media, sinusitis, and acute bronchitis. Also, rhinovirus infections can cause secondary bacterial infections in children, the elderly, and immunosuppressed people. This secondary bacterial infection can cause pneumonia.

As used herein, "treat", "treatment", or "treatment" in relation to a disorder means: (1) to alleviate the disorder or one or more of the biological manifestations of the disorder; (2) show opposition to (a) one or more elements of the biological cascade that leads to or is responsible for the disturbance, or (b) one or more biological manifestations of the disturbance; (3) alleviate one or more of the symptoms or effects associated with the disorder; or (4) slow the progression of the disorder or one or more of the biological manifestations of the disorder.

In the present text, "patient" refers to a human (including adults and children) or other animal. In one embodiment, "patient" refers to a human.

It is contemplated that the compounds of the present invention may be administered topically, for example by inhalation or intranasally. Inhalation refers to the introduction into the lungs of the patient by inhalation through the mouth or through the nasal passages. In one embodiment, the compounds of Formula (I) their pharmaceutically acceptable salts can be administered topically. In another embodiment, the compounds of Formula (I) or their pharmaceutically acceptable salts can be administered by inhalation. In another embodiment, the compounds of Formula (I) or their pharmaceutically acceptable salts can be administered intranasally. In another embodiment, the compounds of Formula (I), or pharmaceutically acceptable salts thereof, may be administered to the eye. In another embodiment, the compounds of Formula (I), or pharmaceutically acceptable salts thereof, may be administered to the ear.

The compounds of the present invention may be administered once a day or according to a dosing regimen in which multiple doses are administered at various intervals over a period of time. For example, doses can be administered one, two, three or four times a day. In one embodiment, the dose is administered once a day. In another embodiment, the dose is administered twice a day. The dose may be administered until the desired therapeutic effect is achieved or indefinitely to maintain the desired therapeutic effect. Suitable administration regimens for a compound of the present invention depend on the pharmacokinetic properties of the compound, such as absorption, distribution and half-life, which can be determined by the skilled artisan. Further suitable administration regimens, including the period during which such regimens are used, for a compound of the present invention depend on the disease being treated, the severity of the disease being treated, the age and physical condition of the patient being treated, the medical history of the patient being treated, the nature concomitant therapy, the desired therapeutic effect, and similar factors that are within the knowledge and experience of a qualified specialist. Further, such skilled artisans will appreciate that suitable dosage regimens may need to be adjusted according to the individual patient's response to a given dosage regimen or over time as the needs of the particular patient change.

Typical daily dosages for administration by inhalation are in the range of 0.2 μg to 0.02 mg per total body weight, eg 0.5 μg to 0.01 mg per total body weight. For example, daily dosages for administration by inhalation may be from 20 μg to 2.0 mg per patient, for example, from 50 μg to 1.0 mg per patient.

Additionally, the compounds of the present invention may be administered as prodrugs. As used herein, a "prodrug" of a compound of the invention is a functional derivative which, when administered to a patient, releases the invention of the invention in vivo. Administration of a compound of the present invention as a prodrug may allow the skilled artisan to do one of the following: (a) modify the onset of activity of the compound in vivo; (b) modify the duration of action of the compound in vivo; (c) modify the transport or distribution of the compound in vivo; (d) change the solubility of the compound in vivo; and (e) overcome side effects or other difficulties associated with the connection. Exemplary functional derivatives useful in the preparation of prodrugs include modifications to the compound that may undergo chemical or enzymatic cleavage in vivo. Such modifications, which include the preparation of phosphates, amides, esters, thioesters and carbamates, are well known to those skilled in the art.

Without wishing to be bound by a particular theory, it is believed that the compounds of the present invention are capable of inhibiting the activity of the host cellular enzyme, PI4KIIIβ, and thereby reduce the ability of the virus to replicate to the host cell. Many viruses use PI4K to generate phosphatidylinositol-4-phosphate (PI4P) enriched membranes that can be used as platforms for replication. The mechanism of viral replication is assembled on these platforms as a supramolecular complex, and PI4P lipids contribute to the synthesis of viral RNA. Such intracellular lipid platforms create a more favorable environment for more efficient virus replication. Disruption of the ability of the virus to use the PI4K and PI4KIIIβ molecules, in particular to create these lipid platforms and promote viral replication, may provide an opportunity for the treatment and/or prevention of viral infections.

Accordingly, according to another aspect of the present invention, there is provided a method for treating a viral infection comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a patient in need thereof.

According to another aspect of the present invention, there is provided a method for treating a disorder caused or aggravated by a viral infection, comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a patient in need thereof.

According to another aspect of the present invention, there is provided a method for treating a secondary bacterial infection caused by a viral infection, comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a patient in need thereof.

According to another aspect of the present invention, there is provided a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for use in therapy.

According to another aspect of the present invention, there is provided a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for use in the treatment of a viral infection.

According to another aspect of the present invention, there is provided a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for use in the treatment of a disorder caused or aggravated by a viral infection. In one embodiment, the disorder is COPD, cystic fibrosis, bronchiectasis, asthma, or congestive heart failure. In another embodiment, the disorder is COPD or asthma. In yet another embodiment, the disorder is COPD.

According to another aspect of the present invention, there is provided a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for use in the treatment of a secondary bacterial infection caused by a bacterial infection.

According to another aspect of the present invention, the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, is provided in the manufacture of a medicament for the treatment of a viral infection.

According to another aspect of the present invention, the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, is provided in the manufacture of a medicament for the treatment of a disorder caused or aggravated by a viral infection.

According to another aspect of the present invention, the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, is provided in the manufacture of a medicament for the treatment of a secondary bacterial infection caused by a viral infection.

For some patients, such as those with a compromised immune system or cardiopulmonary comorbidities who are at increased risk of developing severe illness following a viral infection, prophylactic administration of the compounds of the present invention is contemplated in order to avoid exacerbating the disorder, such as COPD, cystic fibrosis, bronchiectasis, asthma or congestive heart failure. It is clear that "prevention" is not an absolute term. In medicine, "prevention" refers to the prophylactic administration of a drug to greatly reduce the likelihood or severity of a disorder, or to delay the onset of such a disease. Prophylactic administration may be particularly useful in cases where there is an increased risk of infection, such as in the case of HRV during the winter months.

According to another aspect of the present invention, there is provided a method for preventing a viral infection comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a patient in need thereof.

According to another aspect of the present invention, there is provided a method for preventing a disorder caused or aggravated by a viral infection, comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a patient in need thereof. In one embodiment, the disorder is COPD, cystic fibrosis, bronchiectasis, asthma, or congestive heart failure. In another embodiment, the disorder is COPD or asthma. In yet another embodiment, the disorder is COPD.

According to another aspect of the present invention, there is provided a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for preventing viral infection.

According to another aspect of the present invention, there is provided a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for preventing a disorder caused or aggravated by a viral infection. In one embodiment, the disorder is COPD, cystic fibrosis, bronchiectasis, asthma, or congestive heart failure. In another embodiment, the disorder is COPD or asthma. In yet another embodiment, the disorder is COPD.

According to another aspect of the present invention, the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, is provided in the manufacture of a medicament for the prevention of viral infection.

According to another aspect of the present invention, the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, is provided in the manufacture of a medicament for the prevention of a disorder caused or aggravated by a viral infection. In one embodiment, the disorder is COPD, cystic fibrosis, bronchiectasis, asthma, or congestive heart failure. In another embodiment, the disorder is COPD or asthma. In yet another embodiment, the disorder is COPD.

Additionally, it is possible to prophylactically administer a compound of the present invention to healthy individuals if they are expected to be at increased risk of viral infection, such as during a SARS outbreak or in a social or healthcare setting where several residents have been exposed to HRV infection. Accordingly, according to another aspect of the present invention, there is provided a method for preventing a viral infection, comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a person in need thereof.

The embodiments described below relate to each of the above aspects related to medical use. In one embodiment, the virus is a single stranded RNA virus. In yet another embodiment, the virus is a single stranded positive sense strand RNA virus. In one embodiment, the viral infection is HRV (HRV-human rhinovirus), i.e. is an RHF infection. In yet another embodiment, the viral infection is RHF and the disorder caused by the virus is the common cold. In yet another embodiment, the viral infection is RHF, wherein the disorder caused by the virus is bronchiolitis, pneumonia, otitis media, sinusitis, or acute bronchitis. In yet another embodiment, the secondary bacterial infection causes pneumonia. In another embodiment, the disorders exacerbated by the virus are COPD, cystic fibrosis, bronchiectasis, asthma, or congestive heart failure. In yet another embodiment, the RHF is RHF-A. In yet another embodiment, the RHF is RHF-B. In yet another embodiment, the RHF represents an RHF-C failure. In another embodiment, the compound of Formula (I) is administered at the onset of nasal symptoms of human rhinovirus to prevent pulmonary rhinovirus infection, and thus reduce the frequency and severity of asthma exacerbations. In another embodiment, the compound of Formula (I) is administered at the onset of nasal symptoms of human rhinovirus to prevent pulmonary rhinovirus infection, and thus reduce the frequency and severity of COPD exacerbations. In another embodiment, the compound of Formula (I) is administered at the onset of nasal symptoms of human rhinovirus in order to prevent pulmonary rhinovirus infection, and thus reduce the frequency and severity of cystic fibrosis exacerbations. In another embodiment, the compound of Formula (I) is administered at the onset of nasal symptoms of human rhinovirus to prevent pulmonary rhinovirus infection, and thereby reduce the frequency and severity of exacerbations of congestive heart failure.

In one embodiment, the viral infection is a coronavirus infection and the disease or condition is severe acute respiratory syndrome (SARS or SARS).

Lineups

Compounds of the present invention will typically, but not necessarily, be formulated into pharmaceutical formulations prior to administration to a patient. According to another aspect of the present invention, there is provided a pharmaceutical composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. According to another aspect of the present invention, a pharmaceutical composition for the treatment of a viral infection is provided, comprising Formula (I) or a pharmaceutically acceptable salt thereof. According to another aspect of the present invention, there is provided a pharmaceutical composition for the treatment of a disorder caused or aggravated by a viral infection, comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof. According to another aspect of the present invention, a pharmaceutical composition for the treatment of a secondary bacterial infection caused by a viral infection is provided, comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof. According to another aspect of the present invention, a pharmaceutical composition for preventing a viral infection is provided, comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof. According to another aspect of the present invention, there is provided a pharmaceutical composition for preventing a disorder caused or aggravated by a viral infection, comprising Formula (I) or a pharmaceutically acceptable salt thereof.

In the present text, "pharmaceutically acceptable excipient" means a pharmaceutically acceptable material, composition or base involved in giving form or consistency to a pharmaceutical composition. Each excipient must be compatible with the other ingredients of the pharmaceutical formulation when combined to avoid interactions that would substantially reduce the efficacy of the compounds of the present invention when administered to a patient and interactions that could lead to the formation of pharmaceutical formulations that are not pharmaceutically acceptable. In addition, each excipient must, of course, be pharmaceutically acceptable, for example of sufficiently high purity.

It is contemplated that the compounds of the present invention may be administered topically, for example by inhalation, intranasally, transdermally, into the eye or into the ear.

According to another aspect, the present invention relates to a dosage form adapted to be administered to a patient by inhalation, for example, in the form of a composition in the form of a dry powder, aerosol, suspension or solution.

Dry powder formulations for pulmonary delivery by inhalation typically contain a compound of the present invention as a finely divided powder together with one or more pharmaceutically acceptable excipients in the form of finely divided powders. Pharmaceutically acceptable excipients particularly suitable for use in dry powder are known to those skilled in the art and include lactose, starch, mannitol, and mono-, di-, and polysaccharides. Fine powder can be obtained by, for example, micronization and milling. Typically, a pulverized (eg, micronized) compound may have a D ₅₀ value of from about 1 to about 10 microns (eg, as measured by laser diffraction).

The dry powder can be administered to a patient using a Reservoir Dry Powder Inhaler (RDPI) having a reservoir suitable for storing multiple (unmetered doses) drugs in dry powder form. The RDPI is typically contained to dispense each dose of drug from the reservoir to the delivery site. For example, the dosing means may include a dispensing cup that is movable from a first position, where the cup can be filled with drug from a reservoir, to a second position, where a metered dose of drug becomes available to the patient for inhalation.

Dry powder formulations for use in accordance with the present invention may be administered via inhalation devices. By way of example, such devices may include capsules and cartridges of, for example, gelatin, or blisters of, for example, laminated aluminum foil. In various embodiments, each capsule, cartridge, or blister may contain doses of a formulation as described herein. Examples of inhalation devices include devices for metered or multi-dose formulation delivery, including all devices provided herein. As an example, in the case of multi-dose delivery, the formulations may be pre-metered (e.g., as in DISKUS, see GB2242134, US Pat. 2,169,265, US Pat. An example of a single dose device is the ROTAHALER (see GB 2064336). In one embodiment, the DISKUS inhalation device comprises an oblong strip made from a base sheet containing a plurality of depressions spaced along its length and a cover sheet that is peelably connected thereto, thereby forming a plurality of containers, each container contains in cheba an inhalation composition containing compounds, optionally with other excipients and additives described in this document. The peelable connection is an engineered connection and, in one embodiment, the engineering connection is a sealed connection. Preferably, the strip is flexible enough to be rolled up. The cover sheet and base sheet preferably have leading end portions that are not connected to each other, and at least one of the leading end portions is configured to be attached to the winding means. Also in the preferred case, the engineering connection between the base and cover sheets extends over the entire width. The cover sheet may preferably be peeled from the base sheet in a longitudinal direction from the first end of the base sheet.

Formulations in dry powder form may also be presented in an inhalation device which is configured to separate two different components of the composition. Thus, for example, these components are administered at the same time, but stored separately, for example, in separate pharmaceutical formulations, for example, as described in, for example, WO 03/061743 A1 WO 2007/012871 A1, WO2007/068896, as well as in US patent no. 8,113,199, 8,161,968, 8,511,304, 8,534,281, 8,746,242 and 9,333,310.

In one embodiment, the inhalation device configured to separate components is an inhaler having two peelable blister strips, each containing pre-metered doses in blister pockets along its length, e.g., multiple containers within each blister strip, e.g. , ELLIPTA. Such a device has an internal indexing mechanism which, on each actuation of the device, peels open the pocket of each strip and positions the blisters so that each newly exposed dose of each strip is adjacent to a manifold that communicates with the mouthpiece of the device. When the patient inhales through the mouthpiece, each dose is simultaneously withdrawn from the respective pocket into the manifold and drawn through the mouthpiece into the patient's airway. Another device designed to separate different components is the DUOHALER from Innovata. In addition, various designs of inhalation devices provide sequential or separate delivery of the pharmaceutical composition(s) from the device in addition to simultaneous delivery.

Alternatively, the dry powder may be presented in capsules (eg, gelatin or plastic), cartridges, or blister packs for use in a multi-dose dry powder inhaler (MDPI). MDPIs are inhalers in which a drug is contained in a multi-dose package containing (or otherwise carrying) several specific doses (or portions thereof) of the drug. When the dry powder is presented as a blister pack, it contains a plurality of blisters for storing the drug in dry powder form. The blisters are usually arranged in a regular manner to facilitate the release of the drug from them. For example, the blisters may be arranged in a substantially circular pattern on the disc-shaped blister pack, or the blisters may be elongated in shape, such as containing a strip or ribbon. Each capsule, cartridge or blister may, for example, contain 200 μg to 10 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

Aerosols can be obtained by suspending or dissolving a compound of the present invention in a liquefied propellant. Suitable propellants include halocarbons, hydrocarbons and other liquefied gases. Typical examples of propellants include: trichlorofluoromethane (propellant 11), dichlorofluoromethane (propellant 12), dichlorotetrafluoroethane (propellant 114), tetrafluoroethane (HFA-134a), 1,1-difluoroethane (HFA-152a), difluoromethane (HFA-32), pentafluoroethane ( HFA-12), heptafluoropropane (HFA-227a), perfluoropropane, perfluorobutane, perfluoropentane, butane, isobutane and pentane. Aerosols containing a compound of the present invention will typically be administered to a patient via a metered dose inhaler (MDI). Such devices are known to those skilled in the art.

The aerosol may contain additional pharmaceutically acceptable excipients commonly used in combination with MDI, such as surfactants, lubricants, co-solvents and other excipients to improve the physical stability of the formulation, to improve valve performance, to improve solubility, to improve taste.

In another aspect, a pharmaceutical aerosol formulation is provided comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a fluorocarbon or chlorofluorocarbon propellant, optionally in combination with a surfactant and/or a co-solvent.

According to one embodiment, the propellant is selected from 1,1,1,2-tetrafluoroethane, 1,1,1,2,3,3,3-heptafluoro-n-propane, and mixtures thereof.

The compositions according to the present invention can be buffered by adding suitable buffer substances.

Capsules and cartridges for use in an inhaler or insufflator of, for example, gelatin can be made containing a powder inhalation mixture of a compound of the present invention and a powder base such as lactose or starch. Typically, each capsule or cartridge may contain 200 μg to 10 mg of a compound of the present invention. Alternatively, the compound of the present invention may be provided without excipients such as lactose.

The proportion of the compound of the present invention in the topical formulations of the present invention depends on the exact type of formulation to be prepared, but will generally be in the range of 0.01 to 10% by weight. Typically, for most types of drugs, this proportion will be in the range of 0.05 to 1%, eg 0.1 to 0.5%.

Aerosol formulations are preferably formulated such that each metered dose or "puff" of aerosol contains 20 μg to 10 mg, preferably 20 μg to 5 mg, more preferably about 20 μg to 0.5 mg of a compound of the present invention. Administration can be done once a day or several times a day, eg 2, 3, 4 or 8 times, with eg 1, 2 or 3 doses each time. The total daily dose using an aerosol will be in the range of 20 µg to 2.0 mg, eg 50 µg to 1.0 mg. The total daily dose and metered doses delivered by capsules and cartridges in an inhaler/insufflator will typically be twice the doses delivered by aerosol formulations.

In the case of suspension aerosol formulations, the particle size of the particulate drug (e.g. micronized) should be such as to allow substantially all of the drug to be inhaled into the lungs upon administration of the aerosol formulation and will accordingly be less than 100 microns, desirably less than 20 microns and , in particular in the range of 1 to 10 microns, such as 1 to 5 microns, more preferably 2 to 3 microns.

The compositions of the present invention can be prepared by dispersing or dissolving the medicament and the compound of the present invention in the selected propellant in a suitable container, for example, using ultrasound or a high shear mixer. This process is preferably carried out under conditions of controlled humidity.

The chemical and physical stability, as well as the pharmaceutical acceptability of aerosol formulations in accordance with the present invention can be determined using techniques well known to those skilled in the art. Accordingly, for example, the chemical stability of these components can be determined using HPLC analysis, for example, after prolonged storage of the product. Physical stability data can be obtained using other standard methods of analysis, such as, for example, no leakage tests, valve flow analysis (average injection masses per actuation), dose reproducibility analysis (amount of active ingredient per actuation), and analysis spray distribution. The stability of the suspension aerosol formulations of the present invention can be measured using standard techniques, for example, by measuring the particle size distribution of the flocculant using a backscatter light instrument; or by measuring the particle size distribution by the cascade impactor method or the "double impinger" analytical method. As used herein, "double impinger" analysis means "determination of the deposited dose emitted by pressurized inhalation using Apparatus A" as defined in the British Pharmacopoeia 1988, pages A204-207 Appendix XVII C. Such techniques allow the calculation of the "respirable fraction » aerosol formulations. One method used to calculate the "respirable fraction" is based on the "fines fraction", which is the amount of active ingredient collected in the lower chamber of the impinger per actuation, expressed as a percentage of the total amount of active ingredient delivered per actuation, at using the double impinger method described above.

The term "metered dose inhaler" or MDI means a device comprising a container, a fixed lid covering the container, and a composition dosing valve located in the lid. The MDI system includes a suitable channeling device. Suitable ducting devices include, for example, an actuating valve and a cylindrical or cone-shaped passage through which medicament can be delivered from the filled cartridge by means of a metering valve into the patient's nose or mouth, such as a mouthpiece valve.

MDI cartridges typically include a container capable of withstanding the vapor pressure of the propellant used, such as a bottle made of plastic or plastic-coated glass, or preferably a metal container, such as aluminum or aluminum alloy, which may optionally be anodized, varnished and/or plastic ( for example, according to WO 96/32099 incorporated herein by reference, all or some of the internal surfaces are coated with one or more fluorocarbon polymers, optionally in combination with one or more non-fluorocarbon polymers), and said container is closed with a metering valve. The lid can be attached to the container by ultrasonic welding, threading or crimping. The MDIs described herein can be prepared using methods known in the art (eg see Byron, supra, and WO 96/32099). Preferably, the container is equipped with a cap assembly, said cap having a drug-dispensing valve and being clamped in place.

In one embodiment of the present invention, the metal inner surface of the container is coated with a fluoropolymer, preferably mixed with a non-fluoropolymer. According to another embodiment of the present invention, the metal inner surface of the container is coated with a mixture of polytetrafluoroethylene (PTFE) and polyethersulfone (PES) polymers. In a further embodiment according to the present invention, the metal inner surface of the container is completely coated with a mixture of polytetrafluoroethylene (PTFE) and polyethersulfone (PES) polymers.

Dispense valves are designed to deliver a metered amount of formulation with each actuation and include a seal to prevent leakage of propellant through the valve. Said seal may comprise any suitable elastic material such as, for example, low density polyethylene, chlorobutyl, bromobutyl, EPDM, black and white acrylonitrile butadiene rubber, butyl rubber and neoprene. Suitable valves are commercially available from manufacturers well known in the aerosol industry, e.g. Valois, France (e.g. DF10, DF30, DF60), Bespak Pic, UK (e.g. BK300, BK357), and 3M-Neotechnic Ltd, UK (e.g. SPRAYMISER).

In various embodiments, these MDIs may also be used in combination with other structures such as, but not limited to, an outer storage package containing the MDIs, including those described in US Pat. Nos. 6,119,853; No. 6179118; No. 6315112; No. 6352152; No. 6390291; and No. 6,679,374, as well as dose counting devices such as, but not limited to, those described in US Pat.

Conventional mass production methods and equipment well known to those skilled in the art of preparing pharmaceutical aerosols can be used to produce large batches for the commercial production of filled cans. Accordingly, for example, in one mass production method for making suspension aerosol formulations, the metering valve is crimped onto an aluminum container to form an empty can. The particulate medicament is added to the filling vessel and the liquefied propellant, along with optional auxiliaries, is pressurized into the production vessel through the filling vessel. The drug suspension is agitated before returning to the filling apparatus, after which an aliquot of the drug suspension is filled into the can through the metering valve. In one example of a mass production process for making aerosol formulations from solutions, a metering valve is crimped onto an aluminum can to form an empty can. The liquefied propellant, along with optional excipients and dissolved medicament, is pressurized into a production vessel through the filling vessel.

In an alternative process, an aliquot of the liquefied formulation is added to an open canister at temperatures low enough so that the formulation does not evaporate, and then the metering valve is crimped onto the canister.

As a general rule, in batches prepared for pharmaceutical use, each filled canister is checkweighed, coded with the batch number, and packed in a storage tray prior to testing the finished product.

Suspensions and solutions containing a compound of the present invention may also be administered to a patient via a nebulizer. The solvent or suspension agent used for nebulization can be any pharmaceutically acceptable liquid such as water, aqueous saline, alcohols or glycols, e.g. ethanol, isopropyl alcohol, glycerin, propylene glycol, polyethylene glycol, and the like, or mixtures thereof. . For saline solutions, salts are used which exhibit little or no pharmacological activity after administration. For this purpose, both inorganic salts, such as alkali metal or ammonium halide salts, such as sodium chloride, potassium chloride, and organic salts, such as potassium, sodium and ammonium salts of organic acids, such as ascorbic acid, citric acid, can be used. , acetic acid, tartaric acid, etc.

Other pharmaceutically acceptable excipients may be added to the suspension or solution. The compound of the present invention can be stabilized by adding an inorganic acid, such as hydrochloric acid, nitric acid, sulfuric acid and/or phosphoric acid; an organic acid such as ascorbic acid, citric acid, acetic acid and tartaric acid, and the like, a complexing agent such as EDTA or citric acid, and salts thereof; or an antioxidant such as the antioxidant vitamin E or ascorbic acid. These agents may be used alone or together to stabilize a compound of formula (I) or a pharmaceutically acceptable salt thereof. Preservatives such as benzalkonium chloride or benzoic acid and its salts may be added. A surfactant may be added, in particular to improve the physical stability of the suspensions. These surfactants include lecithin, disodium dioctylsulfosuccinate, oleic acid and sorbitol esters.

Formulations for nasal administration may include pressurized aerosol formulations and aqueous formulations that are delivered into the nose using a pressurized pump. In particular, non-pressurized formulations adapted for topical administration in the nasal cavity are of interest. Suitable formulations contain water as a diluent or carrier for this purpose. Aqueous formulations for pulmonary or nasal administration may be provided with standard excipients such as buffering agents, tonicity modifying agents, and the like. Aqueous formulations may also be administered to the nose by nebulization.

The compounds of the present invention may be incorporated into a liquid composition for delivery from a liquid dispenser, e.g., a liquid dispenser with a metering nozzle or metering orifice through which a metered dose of the liquid composition is delivered when force is applied by the user to the pump mechanism of said liquid dispenser. Such liquid dispensers are typically provided with a reservoir of several metered doses of liquid composition which can be dispensed upon successive actuations of the pump. The metering nozzle or orifice may be configured to be placed in the user's nostrils to dispense a spray of the liquid composition into the nasal cavity. A liquid dispenser of the above type is described and illustrated in WO 05/044354, the entire content of which is incorporated herein by reference in its entirety. The dispenser includes a housing, inside which is placed a device for injecting a liquid containing a compression pump mounted on a container for a liquid composition. The housing has at least one finger-operated side lever that is capable of moving inwards relative to the housing, pulling the container inside the housing upwards, as a result of which the pump is compressed and pumps out a measured dose of the composition from the pump rod through the nasal nozzle on the housing. In one embodiment, said liquid dispenser is of the general type shown in FIG. 30-40 WO 05/044354.

Pharmaceutical formulations adapted for intranasal administration, where the carrier is a solid, include a coarse powder with particle sizes, for example, in the range of 20-500 microns, which is administered by rapid inhalation through the nasal canal from a powder container held near the nose. Suitable formulations wherein the carrier is a liquid for administration as a nasal spray or nasal drops include aqueous or oily solutions of a compound of the present invention.

Some of the disorders caused or exacerbated by viral infections can lead to skin problems such as skin rashes. Pharmaceutical formulations adapted for transdermal administration may be presented as individual patches intended to be in close contact with the patient's epidermis for an extended period of time. For example, the active ingredient can be delivered from the patch by iontophoresis, as generally described in Pharmaceutical Research, 3(6), 318 (1986).

Pharmaceutical formulations adapted for topical administration may be formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosols or oils.

Ointments, creams and gels may, for example, be formulated with an aqueous or oily base with the addition of a suitable thickening and/or gelling agent and/or solvents. Such bases may suitably, for example, include water and/or an oil such as liquid paraffin or a vegetable oil such as peanut oil or castor oil, or a solvent such as polyethylene glycol. Thickening agents and gelling agents that may be used depending on the nature of the base include soft paraffin, aluminum stearate, cetostearyl alcohol, polyethylene glycols, wool fat, beeswax, carboxypolymethylene and cellulose derivatives, and/or glyceryl monostearate and/or nonionic emulsifying agents.

Lotions may be formulated with an aqueous or oily base and, in general, also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, or thickening agents.

Powders for topical use can be prepared using any suitable powder base, such as talc, lactose, or starch. Drops may be formulated with an aqueous or non-aqueous base also containing one or more dispersing agents, solubilizing agents, suspending agents or preservatives.

Topical formulations may be administered by one or more applications per day to the affected area. On skin areas, it is preferable to use occlusive dressings. Continuous or sustained delivery can be achieved through the use of an adhesive reservoir system.

For intraocular or otic treatment, the formulations may be applied as a topical ointment or cream. When formulated into an ointment, a compound of formula (I), or a pharmaceutically acceptable salt thereof, may be used with either a paraffinic or water-miscible ointment base. Alternatively, a compound of formula (I), or a pharmaceutically acceptable salt thereof, may be formulated into a cream with an oil-in-water cream base or a water-in-oil base.

Pharmaceutical formulations adapted for parenteral administration include aqueous and non-aqueous sterile injectable solutions that may contain antioxidants, buffers, bacteriostatic agents, and solutes that render the formulation isotonic with the blood of the intended recipient; as well as aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickening agents. These formulations may be provided in unit-dose or multi-dose containers, such as sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) state, requiring only the addition of a sterile liquid carrier, such as water for injection, immediately prior to use. Extemporaneous injectable solutions and suspensions can be prepared from sterile powders, granules and tablets.

Compounds or pharmaceutical formulations of the present invention may be co-administered with an anti-inflammatory agent such as a corticosteroid or pharmaceutical formulation containing the same for the treatment of asthma aggravated by viral infections, in particular HRV infections. For example, the compounds of the present invention may be co-administered with an anti-inflammatory agent such as a corticosteroid in a single formulation, such as a dry powder formulation for inhalation. Alternatively, a pharmaceutical formulation containing a compound of the present invention may be administered in combination with a pharmaceutical formulation containing an anti-inflammatory agent such as a corticosteroid, either simultaneously or sequentially. For example, both pharmaceutical formulations, a pharmaceutical formulation containing a compound of the present invention and an additional pharmaceutical formulation containing an anti-inflammatory agent such as a corticosteroid, may be in a device suitable for simultaneous administration of both formulations via inhalation.

Suitable corticosteroids for administration with the compounds of the present invention include fluticasone furoate, fluticasone propionate, beclomethasone dipropionate, budesonide, ciclesonide, mometasone furoate, triamcinolone, flunisolide, and prednisolone. Suitable corticosteroids for administration with the compounds of the present invention by inhalation include fluticasone furoate, fluticasone propionate, beclomethasone dipropionate, budesonide, ciclesonide, mometasone furoate and flunisolide.

Accordingly, according to a further aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, and one or more anti-inflammatory agents such as a corticosteroid or a phosphatidylinositol-4,5-bisphosphate-3-kinase inhibitor. delta (PI3Kδ).

According to a further aspect, the present invention provides a method of treating asthma aggravated by a viral infection, e.g., HRV, which comprises administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, and one or more anti-inflammatory agents, such as like a corticosteroid.

According to a further aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, and one or more anti-inflammatory agents, such as a corticosteroid, for use in the treatment of asthma aggravated by a viral infection.

According to a further aspect, the present invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, and one or more anti-inflammatory agents such as a corticosteroid, in the manufacture of a medicament for the treatment of asthma aggravated by a viral infection.

According to a further aspect, the present invention provides a method of treating cystic fibrosis aggravated by a viral infection, e.g., HRV, which comprises administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, and one or more anti-inflammatory agents, such as a corticosteroid.

According to a further aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, and one or more anti-inflammatory agents, such as a corticosteroid, for use in the treatment of cystic fibrosis aggravated by a viral infection.

According to a further aspect, the present invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, and one or more anti-inflammatory agents such as a corticosteroid, in the manufacture of a medicament for the treatment of cystic fibrosis aggravated by a viral infection.

According to a further aspect, the present invention provides a method of treating congestive heart failure aggravated by a viral infection, e.g., HRV, which comprises administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, and one or more anti-inflammatory agents. such as a corticosteroid.

According to a further aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, and one or more anti-inflammatory agents, such as a corticosteroid, for use in the treatment of congestive heart failure aggravated by a viral infection.

In a further aspect, the present invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, and one or more anti-inflammatory agents such as a corticosteroid, in the manufacture of a medicament for the treatment of congestive heart failure aggravated by a viral infection.

The following implementation options apply to any of the above aspects related to combinations with one or more anti-inflammatory agents.

In an embodiment, said pharmaceutical composition contains a single anti-inflammatory agent.

In one embodiment, said anti-inflammatory agent is a corticosteroid.

In one embodiment, said corticosteroid is fluticasone furoate.

In one embodiment, said corticosteroid is fluticasone propionate.

In one embodiment, said anti-inflammatory agent is a phosphatidylinositol-4,5-biphosphate-3-kinase delta (PI3Kδ) inhibitor, such as nemiralisib (see Sriskantharajah et al., Annals of the New York Academy of Sciences, (2013) , 1280, 35; Cahn et al., Pulmonary Pharmacology and Therapeutics, (2017), 46, 69; and Stark et al., Current Opinion in Pharmacology, (2015), 23, 82).

The compounds or pharmaceutical formulations of the present invention may be administered in conjunction with one or more bronchodilators, or pharmaceutical formulations containing them, for the treatment of COPD aggravated by a viral infection. For example, the compounds of the present invention may be co-administered with one or more bronchodilators in a single formulation, such as a dry powder formulation for inhalation. Alternatively, a pharmaceutical formulation containing a compound of the present invention may be administered in combination with a pharmaceutical formulation containing one or more bronchodilators, either simultaneously or sequentially. In a further alternative embodiment, a formulation containing a compound of the present invention and a bronchodilator may be administered in combination with a pharmaceutical formulation containing an additional bronchodilator. For example, each of the formulations, a pharmaceutical formulation containing a compound of the present invention and an additional pharmaceutical formulation containing one or more bronchodilators, may be in a device suitable for simultaneous administration of both formulations by inhalation.

Suitable bronchodilators for administration with the compounds of the present invention include β ₂ -adrenergic agonists and anticholinergic agents. Examples of _β2 -adrenergic agonists include, for example, vilanterol, salmeterol, salbutamol, formoterol, salmefamol, fenoterol, carmoterol, ethanterol, naminterol, clenbuterol, pirbuterol, flerbuterol, reproterol, bambuterol, indacaterol, terbutaline and their salts, e.g. xinafoate (1 -hydroxy-2-naphthalenecarboxylate) salt of salmeterol, sulfate salt of salbutamol or fumarate salt of formoterol. Examples of anticholinergic agents include umeclidinium (eg as bromide), ipratropium (eg as bromide), oxitropium (eg as bromide) and tiotropium (eg as bromide). In one embodiment, a compound of the present invention may be co-administered with a β ₂ -adrenergic agonist such as vilanterol and an anticholinergic agent such as umeclidinium.

According to a further aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, and one or more bronchodilators.

According to a further aspect, the present invention provides a method of treating COPD aggravated by a viral infection, which comprises administering to a subject in need a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, and one or more bronchodilators.

According to a further aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, and one or more bronchodilators for use in the treatment of COPD aggravated by a viral infection.

According to a further aspect, the present invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, and one or more bronchodilators in the manufacture of a medicament for the treatment of COPD aggravated by a viral infection.

The following embodiments apply to any of the above aspects relating to combinations with one or more bronchodilators.

In an embodiment, said one or more bronchodilators contain one or more β ₂ -adrenergic receptor agonists.

In an embodiment, said one or more bronchodilators contain one or more anticholinergic agents.

In an embodiment, said one or more bronchodilators comprise one or more β ₂ -adrenergic agonists and one or more anticholinergic agents.

In an embodiment, said one or more bronchodilators comprise a β ₂ -adrenergic agonist and an anticholinergic agent.

In an embodiment, the implementation of these one or more bronchodilators contain one bronchodilator, which is a β ₂ -adrenergic agonist.

In an embodiment, said one or more bronchodilators comprise a single bronchodilator that is an anticholinergic agent.

In an embodiment, said β ₂ -adrenergic agonist is vilanterol.

In an embodiment, said anticholinergic agent is umeclidinium. In a further embodiment, said anticholinergic agent is umeclidinium bromide.

According to a further aspect, the present invention provides a pharmaceutical composition comprising a) a compound of formula (I) or a pharmaceutically acceptable salt thereof, b) one or more bronchodilators; and c) one or more anti-inflammatory agents.

According to a further aspect, the present invention provides a method of treating COPD aggravated by a viral infection, which comprises administering to a subject in need thereof a therapeutically effective amount of a) a compound of formula (I) or a pharmaceutically acceptable salt thereof, b) one or more bronchodilators; and c) one or more anti-inflammatory agents.

According to a further aspect, the present invention provides a) a compound of formula (I) or a pharmaceutically acceptable salt thereof, b) one or more bronchodilators; and c) one or more anti-inflammatory agents for use in the treatment of COPD aggravated by a viral infection.

According to a further aspect, the present invention provides the use of a) a compound of formula (I) or a pharmaceutically acceptable salt thereof, b) one or more bronchodilators; and c) one or more anti-inflammatory agents in the manufacture of a medicament for the treatment of COPD aggravated by a viral infection.

The following embodiments apply to any of the above aspects relating to combinations with one or more bronchodilators and one or more anti-inflammatory agents.

In an embodiment, said one or more bronchodilators contain one or more β ₂ -adrenergic receptor agonists.

In an embodiment, said one or more bronchodilators contain one or more anticholinergic agents.

In an embodiment, said one or more bronchodilators comprise one or more β ₂ -adrenergic agonists and one or more anticholinergic agents.

In an embodiment, said one or more bronchodilators comprise a β-adrenergic agonist and an anticholinergic agent.

In an embodiment, the implementation of these one or more bronchodilators contain one bronchodilator, which is a β ₂ -adrenergic agonist.

In an embodiment, said one or more bronchodilators comprise a single bronchodilator that is an anticholinergic agent.

In an embodiment, said β ₂ -adrenergic agonist is vilanterol.

In an embodiment, said anticholinergic agent is umeclidinium. In a further embodiment, said anticholinergic agent is umeclidinium bromide.

In an embodiment, said one or more anti-inflammatory agents is a corticosteroid.

In a further embodiment, said corticosteroid is fluticasone furoate.

In a further embodiment, said corticosteroid is fluticasone propionate.

In a further embodiment, said one or more bronchodilators are vilanterol and umeclidinium.

In a further embodiment, said one or more bronchodilators are vilanterol and umeclidinium and said one or more anti-inflammatory agents are fluticasone furoate.

In a further embodiment, said one or more bronchodilators are vilanterol and umeclidinium and said one or more anti-inflammatory agents are fluticasone propionate.

Compounds of the present invention may have an improved profile over known PI4KIIIβ inhibitors, for example, over known PI4KIIIβ inhibitors, certain compounds of the present invention may have one or more of the following properties:

(i) more potent PI4KIIIβ inhibitory activity;

(ii) improved selectivity for PI4KIIIβ;

(iii) an increase in T _1/2 of the enzyme;

(iv) improved efficiency in cells;

(v) Improved lung retention

(vi) improved solubility; and/or

(vii) lower levels of compound accumulation in body tissues.

As noted above, levels of the compounds of the present invention may accumulate in body tissues after administration, lower than those of known PI4KIIIβ inhibitors. In particular, the compounds of the present invention may have lower levels of accumulation in the spleen. Previous studies have shown that the accumulation of PI4KIIIB inhibitors in the spleen may be unfavorable, for example, may induce apoptosis. Confirming Connections

The supporting compounds below illustrate the invention as a guide to the preparation and use of the compounds, compositions and methods of the invention for the skilled artisan. While specific embodiments of the invention have been described, the skilled artisan will appreciate that various changes and modifications may be made. Indications of preparation procedures carried out similarly to other preparation procedures or according to a general method may cover changes in standard parameters such as time, temperature, processing conditions, changes in the amounts of reagents, etc.

Reactions using metal hydrides (including sodium hydride) and organometallic reagents are carried out under argon or nitrogen, unless otherwise indicated.

In the intermediates described below and confirming compounds for which the relative stereochemistry of the compound has been identified, this is indicated both in the name and in the structure of the compound.

For some of the intermediates and confirming compounds described below, starting materials are identified by reference to other intermediate or compound numbers of the invention. This does not mean that the actual material (or "batch") obtained from any particular intermediate or confirming compound was necessarily used in the subsequent step exemplified herein.

Unless otherwise noted, source materials were available for purchase. All solvents and commercial reagents were of laboratory quality and were used unchanged.

). Там, где абсолютная стереохимия неизвестна, но известно, что это единственный энантиомер, используется звездочка (*).If the absolute stereochemistry is known and the compound is a single enantiomer, bold or shaded wedges are used where necessary (

). Where the absolute stereochemistry is not known, but it is known to be the only enantiomer, an asterisk (*) is used.

The names of the intermediates and confirming compounds were generated using the naming program in "ChemBioDraw Ultra v12" or "ACD Name Pro 6.02".

As used herein, the symbols and conventions used in these processes, schemes, and examples are consistent with those used in current scientific literature, such as the Journal of the American Chemical Society. Abbreviations

The list below defines some of the abbreviations and symbols used in this document. It should be understood that the list is not exhaustive, but the meaning of those abbreviations and symbols that are not defined hereinafter is clear to those skilled in the art. In the description of the invention, the chemical elements are identified in accordance with the periodic table of elements.

Bu - butyl

m-CPBA - meta-chloroperoxybenzoic acid

CV - Column volume(s)

DXM - Dichloromethane

DIBAL-H - Diisobutylaluminum hydride

DIPEA - N,N-diisopropylethylamine

DMAP - 4-dimethylaminopyridine

DMP - Dess-Martin Periodinan

DMF - N,N-dimethylformamide

DMSO - Dimethyl Sulfoxide

Dppf - 1,1'bis(diphenylphosphino)ferrocene

HATU - 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate

HCI - Hydrochloric acid

IPA - Isopropanol

2-methyltetrahydrofuran - 2-methyltetrahydrofuran

BSI - N-bromosuccinimide

N-iodosuccinimide - N-iodosuccinimide

rt(v.sp.) - Retention time

TFA - Trifluoroacetic acid

THF - Tetrahydrofuran

HPLC - High Performance Liquid Chromatography

MDAP - Mass Directed Autopreparative HPLC

XPhos - dicyclohexyl(2',4',6'-triisopropyl-[1,1'-biphenyl]-3-yl)phosphane

XPhos Pd G2 - chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II)

LCMS Methods

Method A

Column: Acquity BEN C ₁₈ (50 mm × 2.1 mm, 1.7 µm). Mobile phase: A: 0.1% formic acid in water; B: 0.1% formic acid in acetonitrile. Time (min)/% B: 0 / 3, 0.4 / 3, 2.0 / 98, 3.4 / 98, 3.5 / 3, 4.0 / 3. Column temperature: 35°C, Flow rate: 0.6 ml/min.

Method B

Column: XBridge VEN C ₁₈ (50 mm × 4.6 mm, 2.5 μM). Mobile phase: A: 5 mM ammonium bicarbonate; B: acetonitrile. Time (min)/% V: 0 / 5, 0.5 / 5, 1 / 15, 3.3 / 98, 5.2 / 98, 5.5 / 5, 6.0 / 5. Column temperature: 35°C, Flow rate: 1.3 ml/min .

Method C

Column: Acquity BEN C ₁₈ (50 mm × 2.1 mm, 1.7 µm). Mobile phase: A: 0.1% formic acid in water; B: 0.1% formic acid in acetonitrile. Time (min)/% V: 0 / 3, 0.4 / 3, 2.5 / 98, 3.4 / 98, 3.5 / 3, 4.0 / 3. Column temperature: 35°C, Flow rate: 0.6 ml/min.

Method D

Column: XBridge VEN C ₁₈ (50 mm × 4.6 mm, 2.5 µm). Mobile phase: A: 5 mM ammonium bicarbonate; B: acetonitrile. Time (min)/% V: 0 / 5, 1.5 / 15, 7 / 98, 9 / 98, 9.5 / 5, 10/5. Column temperature: 35°C, Flow rate: 1.3 ml/min.

Method E

Column: Acquity BEN C ₁₈ (50 mm × 2.1 mm, 1.7 µm). Mobile phase: A: 0.05% formic acid in water; B: 0.05% formic acid in acetonitrile. Time (min)/% B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3. Column temperature: 35°C, Flow rate: 0.6 ml/min.

Method F

Column: Acquity UPLC CSH C ₁₈ column (50 mm × 2.1 mm, i.d. 1.7 µm) at 40°C. The following solvents were used: A=10 mM ammonium bicarbonate in water adjusted to pH 10 with ammonia solution. B = cetonitrile. Time (min)/% V: 0/3, 0.05/3, 1.5/95, 1.9/95, 2.0/3. Flow rate: 1 ml/min. MS: Waters ZQ. Ionization mode: Scanning mode positive / negative electrospray

Method G

Column: Acquity BEN C ₁₈ (50 mm × 2.1 mm, 1.7 µm). Mobile phase: A: 0.1% formic acid in water; B: 0.1% formic acid in acetonitrile. Time (min)/% B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3. Column temperature: 35°C, Flow rate: 0.6 ml/min.

Method H

Column: Acquity BEN C ₁₈ (50 mm × 2.1 mm, 1.7 µm). Mobile phase: A: 5 mM ammonium bicarbonate in water (pH 10); B: acetonitrile. Time (min)/% V: 0 / 3, 0.4 / 3, 2.5 / 98, 3.4 / 98, 3.5 / 3, 4.0 / 3. Column temperature: 35°C, Flow rate: 0.6 ml/min.

Method I

Column: Acquity BEN C ₁₈ (100 mm × 2.1 mm, 1.7 µm). Mobile phase: A: 0.05% TFA in water; B: acetonitrile. Time (min)/% V: 0 / 3, 0.4 / 3, 3.5 / 98, 4.5 / 98, 5.0 / 3, 5.5 / 3. Column temperature: 35°C, Flow rate: 0.45 ml/min.

Method J

Column: Acquity UPLC CSH C ₁₈ (50 mm × 2.1 mm, i.d. 1.7 µm) at 40°C. The following solvents were used: A=0.1% v/v. solution of formic acid in water. B=0.1% v/v solution of formic acid in acetonitrile. Time (min)/% V: 0/3, 1.5/95, 1.9/95, 2.0/3. Flow rate: 1 ml/min. MS: Waters ZQ. Ionization Mode: Scanning positive/negative electrospray mode.

Method K

Column: XSelect CSH C ₁₈ (150 mm × 3.0 mm, 2.5 µm). Mobile phase: A: 0.05% TFA in water; B: 100% acetonitrile. Time (min)/% V: 0 / 3, 1 / 3, 8 / 98, 11 / 98, 11.1 / 3, 12 / 3. Temp. columns: 35°C, Flow rate: 0.7 ml/min.

Method L

Column: BEN C ₁₈ (100 mm × 2.1 mm, 1.7 µm). Mobile phase: A: 0.1% TFA in water, B: OI % TFA in acetonitrile. Time (min)/% V: 0 / 3, 8.5 / 100, 9.0 / 100, 9.5 / 3, 10 / 3. Temp, columns: 50°C, Flow rate: 0.55 ml/min.

Method M

Column: CSH C ₁₈ (100 mm × 2.1 mm, i.d. 1.7 µm). Mobile phase: A: 0.1% v/v a solution of formic acid in water; B: 0.1% v/v solution of formic acid in acetonitrile. Time (min)/% V: 0/3, 8.5/99.9, 9/99.9, 9.5/3, 10/3. Temp, columns: 50°C. Flow rate: 0.8 ml/min.

Mass Directed Automated Preparative HPLC (MDAP)

Methods for mass directed automated preparative HPLC used to purify compounds are described below. Solvent gradients were used to elute from 0 to 99% Solvent B in Solvent A over time periods up to 25 minutes. For all methods (unless otherwise noted):

Detection using a diode array detector was carried out at a wavelength from 210 nm to 350 nm. MS Conditions: MS: Waters ZQ

Ionization mode: Positive/negative electrospray scanning mode Scanning range: 100-1000 amu Scan time: 0.2s or 0.50s. Interval between measurements: 0.1 s or 0.2 s

Injection volume: 1ml or 3ml

Method A

Column: Xselect CSH C ₁₈ (150 mm × 30 mm i.d. 5 µm fill diameter) at ambient temperature. The following solvents were used: A=10 mM ammonium bicarbonate adjusted to pH 10 with ammonia in water. B = acetonitrile. Flow rate: 40 ml/min.

Method B

Column: Xselect CSH C ₁₈ column (150 mm × 30 mm, 5 µm id - fill diameter) at ambient temperature. The following solvents were used:

A=0.1% v/v formic acid solution in water

B=0.1% v/v solution of formic acid in acetonitrile.

Flow rate: 40 ml/min.

Intermediate 1

2-bromo-1-methyl-1H-imidazole

1-Methyl-1H-imidazole (20 g, 244 mmol) was dissolved in anhydrous THF (200 ml) under nitrogen atmosphere. The solution was stirred at -78°C and added dropwise n-BuLi (167 ml, 268 mmol) slowly at -78°C. After 1 h, a solution of CBr ₄ (97 g, 292 mmol) in anhydrous THF (200 ml) was added. The solution was stirred for 2 hours at -78°C and for 1 hour at room temperature. The reaction mixture was quenched with saturated aqueous ammonium chloride (300 ml), extracted with ethyl acetate (2×200 ml), dried using anhydrous sodium sulfate, filtered and evaporated under reduced pressure. The crude compound was purified by silica gel chromatography eluting with 60% ethyl acetate in petroleum ether to give the title compound. LCMS (method B): retention time=1.89, [M+H] ⁺ =161.

Intermediate 2

3-(1-Methyl-1H-imidazol-2-yl)benzonitrile

A mixture of (3-cyanophenyl)boronic acid (16 g, 109 mmol), 2-bromo-1-methyl-1H-imidazole (intermediate 1, 15.8 g, 98 mmol) and sodium carbonate (46.2 g, 436 mmol) in isopropyl alcohol (120 mL) and water (120 mL) were stirred and degassed with nitrogen for ₂₀ min; test tube. After cooling, the reaction mixture was diluted with ethyl acetate (300 ml), washed with water (300 ml), dried using anhydrous sodium sulfate, filtered and evaporated under reduced pressure. The crude compound was purified by silica gel chromatography eluting with 70% ethyl acetate in petroleum ether to give the title compound. LCMS (method C): retention time=0.57, [M+H] ⁺ =184.

Intermediate 3

(3-(1-methyl-1H-imidazol-2-yl)phenyl)methanamine

Raney nickel (5 g, 65.5 mmol) was added to a mixture of 3-(1-methyl-1H-imidazol-2-yl)benzonitrile (intermediate 2, 12 g, 65.5 mmol) in 7 M ammonium in methanol (150 ml) at 0°C. The reaction mixture was stirred under hydrogen pressure (60 psi at room temperature, for 24 h. The reaction mixture was filtered through a pad of celite, washed with methanol (300 ml) and the filtrate was concentrated under reduced pressure. The reaction was then repeated on the same scale. The two batches of crude material were combined and purified by neutral alumina chromatography to give the title compound LCMS (Method D): retention time=2.91, [M+H] ⁺ =188.

Intermediate 4

Ethyl 7-hydroxy-2-methylpyrazolo[1,5-а]pyrimidine-5-carboxylate

To a solution of (Z)-1,4-diethoxy-1,4-dioxobut-2-en-2-olate sodium (21.64 g, 103 mmol) in ethanol (100 ml) was added 4 M HCI in 1,4 dioxane (28.3 ml) and then 5-methyl-1H-pyrazole-3-amine (10 g, 103 mmol) at 0°C. The reaction mixture was heated to 85° C. and stirred for 3 hours. The reaction mixture was cooled to room temperature and concentrated. The residue was dissolved in 10% methanol in DCM (200 ml) and washed with saturated sodium bicarbonate solution (100 ml). The organic layer was washed with brine (50 ml), dried using anhydrous sodium sulfate, filtered and evaporated to dryness under reduced pressure. The residue was triturated with diethyl ether to give the title compound. LCMS (method A): retention time=1.53, [M+H] ⁺ =222.

Intermediate 5

Ethyl 7-chloro-2-methylpyrazolo[1,5-а]pyrimidine-5-carboxylate

Phosphorus oxychloride (50 ml, 536 mmol) was added to ethyl 7-hydroxy-2-methylpyrazolo[1,5-a]pyrimidine-5-carboxylate (intermediate 4, 10 g, 45.2 mmol) and the reaction mixture was stirred for 24 h at 90°C. The reaction mixture was cooled to room temp. and concentrated under reduced pressure. The residue was dissolved in DCM (100 ml) and the pH was adjusted to neutral with saturated sodium bicarbonate solution (60 ml). The organic layer was washed with water (100 ml), brine (100 ml), dried using sodium sulfate, filtered and concentrated. The residue was dissolved in 10% methanol in DCM (100 ml) and adsorbed onto silica gel (14 g), then purified by chromatography on silica gel (50 g) eluting with 10% ethyl acetate in petroleum ether. Product-containing fractions were combined and evaporated under reduced pressure to give the title compound. LCMS (method B): retention time=3.13, [M+H] ⁺ =240.

Intermediate 6

Ethyl 7-chloro-3-iodo-2-methylpyrazolo[1.5-а]pyrimidine-5-carboxylate

Acetic acid (25 ml) and N-iodosuccinimide (14.79 g, 65.7 mmol). The reaction mixture was stirred for 6 h at 28°C. The reaction mixture was diluted with dichloromethane (150 mL), washed with saturated sodium thiosulfate (2 x 90 mL), brine (60 mL), dried over sodium sulfate, filtered and concentrated to give the title compound. LCMS (method A): retention time=2.24, [M+H] ⁺ =366.

Intermediate 7

Ethyl 3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidine-5-carboxylate

To a solution of DIPEA (14.33 ml, 82 mmol) in DMSO (50 ml) was added (3-(1-methyl-1H-imidazol-2-yl)phenyl)methanamine (intermediate 3, 5.38 g, 28.7 mmol) and ethyl 7 -chloro-3-iodo-2-methylpyrazolo[1,5-a]pyrimidine-5-carboxylate (intermediate 6, 10 g, 27.4 mmol) and the reaction mixture was stirred for 16 h. The reaction mixture was poured into ice water (200 ml) and the solid was filtered, washed with water (50 ml) and dried in vacuo to give the title compound. LCMS (method A): retention time=1.79, [M+H] ⁺ =517.

Intermediate 8

4-bromo-1-methoxy-2-(methylsulfonyl)benzene

To a stirred solution of sodium sulfite (16.55 g, 131 mmol) and sodium bicarbonate (11.03 g, 131 mmol) in water (160 ml) was added 5-bromo-2-methoxybenzene-1-sulfonyl chloride (25 g, 88 mmol) in 1, 4-dioxane (160 ml) at 70°C. The reaction mixture was stirred at 70° C. for 1 hour. The reaction mixture was cooled to room temp. and the solvent was removed under reduced pressure to give the title crude intermediate as a white solid. The crude product was dissolved in DMF (300 ml) and methyl iodide (10.95 ml, 175 mmol) was added at room temp. and the mixture was stirred at room temp. over 2 hours. The reaction mixture was poured into ice water, extracted with ethyl acetate (300 ml), dried using anhydrous sodium sulfate, filtered and dried under reduced pressure. The crude compound was washed with n-pentane (100 ml) to give the title compound. LCMS (method A): retention time=1.84, [M+H] ⁺ =265.

Intermediate 9

2-(4-methoxy-3-(methylsulfonyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

To a stirred solution of 4-bromo-1-methoxy-2-(methylsulfonyl)benzene (intermediate 8, 15 g, 56.6 mmol) and 4,4,4',4',5,5,5',5-octamethyl- 2,2'-bi(1,3,2-dioxaborolane) (21.55 g, 85 mmol) in 1,4-dioxane (150 ml) was added potassium acetate (8.33 g, 85 mmol) at room temp. The reaction mixture was degassed with argon for 15 min, then the PdCI2(dppf)-DXM adduct (2.31 g, 2.83 mmol) was added to the reaction mixture at room temperature. and again degassed for 15 min under argon. The reaction mixture was stirred at 100° C. for 3 hours. After cooling, the reaction mixture was filtered through a Celite pad, washed with methanol (50 ml) and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography eluting with 20% ethyl acetate in hexane to give the title compound. LCMS (method E): retention time=2.16, [M+H] ⁺ =313.

Intermediate 10

Ethyl 3-(4-methoxy-3-(methylsulfonyl)phenyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidine-5-carboxylate

2-(4-methoxy-3-(methylsulfonyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (intermediate 9, 680 mg, 2.179 mmol), ethyl 3-iodo-2 -methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidine-5-carboxylate (intermediate 7, 750 mg, 1.453 mmol) , chlorine(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (57.1 mg, 0.073 mmol) and potassium fluoride (253 mg, 4.36 mmol) were dissolved in 1,4-dioxane (10 ml) and water (5 ml) in a microwave treatment vessel, the vessel was sealed and degassed with nitrogen. The reaction mixture was stirred at 80° C. for 16 h. '-biphenyl)]palladium(H) (57.1 mg, 0.073 mmol), potassium fluoride (253 mg, 4.36 mmol) and 2-(4-methoxy-3-(methylsulfonyl)phenyl)-4,4,5,5- tetramethyl-1,3,2-dioxaborolane (intermediate compound 9, 680 mg, 2.179 mmol) and the reaction mixture was stirred for another 8 h at 80°C. Then chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium (II) (57.1 mg , 0.073 mmol), potassium fluoride (253 mg, 4.36 mmol) and 2-(4-methoxy-3-(methylsulfonyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (intermediate 9, 680 mg, 2.179 mmol) and the reaction mixture was stirred for another 16 h at 80°C. The reaction mixture was cooled to room temperature, filtered through celite (washing with 3 x 20 ml ethyl acetate) and the solvent was removed under vacuum. The reaction mixture was partitioned between ethyl acetate (50 ml) and water (20 ml) and the organic phase was separated, dried with a hydrophobic filter and the solvent was removed under vacuum. Purification by chromatography on silica gel (120 g) eluting with 0-100% ethyl acetate in cyclohexane (over 20 column volumes) gave the title compound. LCMS (method F): retention time=1.04, [M+H] ⁺ =515.

Intermediate 11

(3-(4-methoxy-3-(methylsulfonyl)phenyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidin-5-yl)methanol

Ethyl 3-(4-methoxy-3-(methylsulfonyl)phenyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidine-5-carboxylate (intermediate 10, 180 mg, 0.313 mmol) was dissolved in THF (5 mL) under nitrogen at 0°C and lithium aluminum hydride (1 M in THF) (0.626 mL, 0.626 mmol) was added, why the reaction mixture was stirred for another 3 h. The reaction was quenched by adding 2M sodium hydroxide solution (5 ml) and stirred for another 1 h. The organic phase was extracted with ethyl acetate (3 × 10 ml), the organic phase was dried using a hydrophobic filter and removed solvent under vacuum to give the title compound. LCMS (method F): retention time=0.89, [M+H] ⁺ =533.

Intermediate 12

3-(4-methoxy-3-(methylsulfonyl)phenyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidine-5-carbaldehyde

(3-(4-methoxy-3-(methylsulfonyl)phenyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidin-5-yl)methanol (intermediate 11, 112 mg, 0.21 mmol) was dissolved in DCM (2.5 mL), manganese dioxide (32 mg, 0.368 mmol) was added, and the reaction mixture was stirred at room temp. for 4 h. Then manganese dioxide (183 mg, 2.103 mmol) was added and the reaction mixture was stirred at room temp. for an additional 16 hours. The reaction mixture was filtered through a Celite cartridge, rinsing with dichloromethane (3 x 5 mL) and the solvent removed in vacuo to give the title compound. LCMS (method F): retention time=1.0, [M+H] ⁺ =531.

Intermediate 13

Ethyl 7-((tert-butoxycarbonyl)(3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)-3-iodo-2-methylpyrazolo[1,5-a]pyrimidine-5-carboxylate

To a solution of ethyl 2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidine-5-carboxylate (intermediate 7, 3.8 g , 9.73 mmol) in DCM (25 ml) was added DIPEA (5.1 ml, 29.2 mmol), DMAP (1.189 g, 9.73 mmol) and Boc anhydride (3.39 ml, 14.6 mmol) and the reaction mixture was stirred for 16 h at room temperature . The reaction mixture was diluted with dichloromethane (50 ml), washed with water (25 ml), dried using sodium sulfate, filtered and concentrated. The residue was dissolved in DCM (30 ml), absorbed onto silica gel (5 g) and purified by chromatography on silica gel (15 g) eluting with ethyl acetate in petroleum ether to give the title compound. LCMS (method d): retention time=2.14, [M+H] ⁺ =617.

Intermediate 14

tert-Butyl (5-(hydroxymethyl)-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H-imidazol-2-yl)benzyl)carbamate

To a solution of ethyl 7-((tert-butoxycarbonyl)(3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)-3-iodo-2-methylpyrazolo[1,5-a]pyrimidine-5- carboxylate (intermediate 13, 500 mg, 0.811 mmol) in ethanol (10 mL), sodium borohydride (92 mg, 2.433 mmol) was added and the reaction mixture was stirred for 3 h at 10°C. The reaction mixture was quenched using 1 M HCl (5 ml), the pH was adjusted to 6, then concentrated. The residue was dissolved in ethyl acetate (25 ml), washed with water (20 ml), dried over sodium sulfate, filtered and concentrated to give the title compound. LCMS (method H): retention time=2.01, [M+H] ⁺ =573.

Intermediate 15

tert-Butyl(5-formyl-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H-imidazol-2-yl)benzyl)carbamate

DMP (1752 mg, 4.13 mmol) was added in one portion to a stirred solution of tert-butyl (5-(hydroxymethyl)-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl) methyl-1H-imidazol-2-yl)benzyl)carbamate (intermediate 14, 1306 mg, 2.274 mmol) in DCM (7.5 ml). The reaction mixture was allowed to stir for 18 hours. Saturated aqueous sodium sulfite solution (10 ml) was added and the reaction mixture was stirred for 2 hours at room temp. The reaction mixture was partitioned between DCM (150 ml) and water (150 ml). The organic layer was separated and the aqueous layer was extracted with additional DCM (2×50 ml). The combined organic layers were washed with saturated aqueous sodium bicarbonate (100 ml), water (100 ml), passed through a hydrophobic sintered filter and evaporated under vacuum. The residue was dissolved in a minimum amount of DCM and purified by silica gel chromatography (120 g) eluting with 0-50% ethyl acetate:ethanol (3:1, v/v) in cyclohexane to give the title compound. LCMS (method F): retention time=1.27, [M+H] ⁺ =573.

Intermediate 16

1-methoxy-2-(methylsulfonyl)benzene

To a stirred solution of (2-methoxyphenyl)(methyl)sulfane (50 g, 324 mmol) in DCM (1 L) at 0° C. under nitrogen was added m-CPBA (140 g, 810 mmol). The reaction mixture was allowed to stir at room temperature for 2 hours. The reaction mixture was diluted with dichloromethane (300 ml), washed with saturated aqueous sodium carbonate solution (2×500 ml), the aqueous phase was extracted with dichloromethane (200 ml). The combined organics were dried using anhydrous sodium sulfate, filtered and evaporated under reduced pressure to give the title compound. LCMS (method I): retention time=2.28, [M+H] ⁺ =187.

Intermediate 17

2-((2-methoxyphenyl)sulfonyl)ethanol

n-Butyl lithium (115 ml, 184 mmol ) and stirred the reaction mixture at -78°C for 30 min, then cooled to 0°C. Paraformaldehyde (73.6 g, 2450 mmol) was added in portions, then the reaction mixture was stirred at 0°C for 2 h. The reaction mixture was quenched with ammonium chloride solution (120 ml), extracted with ethyl acetate (2 × 200 ml). The combined organics were washed with brine (150 ml) and dried using anhydrous sodium sulfate, filtered and evaporated under reduced pressure. The crude compound was pre-absorbed onto silica gel (4 g) and purified by silica gel chromatography (10 g column, 80% ethyl acetate in hexane) to give the title compound. LCMS (method C): retention time=1.16, [M+H] ⁺ =217.

Intermediate 18

2-((5-bromo-2-methoxyphenyl)sulfonyl)ethanol

Recrystallized BSI (16.46 g, 92 mmol) was added to a stirred solution of 2-((2-methoxyphenyl)sulfonyl)ethanol (intermediate 17, 10 g, 46.2 mmol) in DMF (100 mL) under nitrogen at room temperature. The reaction mixture was heated to 50° C. for 18 hours. The reaction mixture was cooled to room temperature and diluted with ethyl acetate (300 ml) and washed with ice water (2×500 ml). The organic layer was separated and the aqueous layer was re-extracted with ethyl acetate (300 ml). The combined organic layers were washed with brine (300 ml) and dried using sodium sulfate, filtered and concentrated under reduced pressure. The crude compound was pre-absorbed onto silica gel (10 g) and purified by silica gel chromatography (90 g column, 40% ethyl acetate in hexanes) to give the title compound. LCMS (method C): retention time=1.85, [M+H] ⁺ =295.

Intermediate 19

2-((2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)sulfonyl)ethanol

To a stirred solution of 2-((5-bromo-2-methoxyphenyl)sulfonyl)ethanol (intermediate 18, 10 g, 33.9 mmol) in 1,4-dioxane (150 ml) 5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (17.2 g, 67.8 mmol) and potassium acetate (9.98 g, 102 mmol) at room temperature, then the reaction mixture was degassed with nitrogen for 15 min. The PdCl ₂ (dppf)-DCM adduct (2.77 g, 3.39 mmol) was added and the reaction mixture was heated at 100°С for 3 h in a sealed tube. The reaction mixture was filtered over a pad of celite and washed with ethyl acetate (2×100 ml) of celite. The combined organics were concentrated under reduced pressure. The reaction was then repeated on the same scale. Two batches of the crude product were mixed and dissolved in DCM (100 ml). The mixture was treated with charcoal (5 g) and refluxed for 10 min before being filtered over a pad of celite. Celite was washed with dichloromethane (2×100 ml). The combined organics were concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate in petroleum ether) to give the title compound. LCMS (method C): retention time=2.17, [M+H] ⁺ =343.

Intermediate 20

tert-Butyl(5-formyl-3-(3-((2-hydroxyethyl)sulfonyl)-4-methoxyphenyl)-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl -1H-imidazol-2-yl)benzyl)carbamate

7-tert-butyl(5-formyl-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H-imidazol-2-yl) benzyl) carbamate (intermediate 15, 487 mg, 0.851 mmol), 2-((2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl) sulfonyl)ethan-1-ol (intermediate 19, 415 mg, 1.03 mmol), PdCl ₂ (dppf) (64 mg, 0.087 mmol) and potassium fluoride (151 mg, 2.6 mmol) in 1,4-dioxane (2 ml ) and water (1 ml). The reaction vessel was sealed and heated in a microwave reactor at 100° C. for a total of 1.5 hours. The reaction mixture was passed through celite, washed with methanol (50 ml) and evaporated under vacuum. The residue was taken up in DCM (30 ml) and partitioned with water (15 ml). The organic layer was separated and the aqueous phase was extracted with additional DCM (2×20 ml). The combined organic layers were passed through a hydrophobic sintered filter and evaporated in vacuo. The residue was purified by chromatography on silica gel (80 g) eluting with 50-100% ethyl acetate:ethanol (3:1, v/v containing 1% triethylamine) in cyclohexane (20 column volumes) to give the title connection header. LCMS (method F): retention time=1.06, [M+H] ⁺ =661.

Intermediate 21

2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a1pyrimidin-5-yl)propan-2- ol

Ethyl 3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidine-5-carboxylate (intermediate 7 , 275 mg, 0.533 mmol) was stirred in THF (5 ml) at 0°С under nitrogen atmosphere. Methylmagnesium bromide (1 M in dibutyl ether) (1.75 ml, 1.75 mmol) was added over 2 minutes and the mixture was allowed to warm to room temperature, after which it was left for 4.5 s. The reaction mixture was quenched with water (2 ml) then separated with dichloromethane (50 ml) and water (50 ml). The organic phase was collected and the aqueous was washed with dichloromethane (50 ml) and then with ethyl acetate (50 ml). The combined organic layers were passed through a hydrophobic sintered filter and evaporated to dryness. The residue was loaded into DCM (2 ml) on a silica gel (40 g) column and eluted with 10-60% ethyl acetate:ethanol (3:1, v/v containing 1% triethylamine) in cyclohexane to give the title connection header. LCMS (method J): retention time=0.7, [M+H] ⁺ =503.

Intermediate 22

5-bromo-2-xpor-N-(3-hydroxypropyl)-N-methylbenzamide

To a stirred solution of 5-bromo-2-chlorobenzoic acid (12.5 g, 53.1 mmol) in THF (100 ml) was added 3-(methylamino)propan-1-ol (5.21 g, 58.4 mmol), DIPEA (27.8 ml, 159 mmol ) and HATU (22.2 g, 58.4 mmol) and the reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was diluted with water (100 ml) and extracted with ethyl acetate (200 ml). The organic layer was dried with anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was dissolved in DCM (50 ml), pre-absorbed onto silica gel (50 g) and purified by silica gel chromatography (150 g, 100% ethyl acetate) to give the title compound. LCMS (method K): retention time=5.30, [M+H] ⁺ =306.

Intermediate 23

2-Chloro-N-(3-hydroxypropyl)-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (9.69 g, 38.2 mmol), potassium acetate (9.60 g, 98 mmol) and degassed reaction mixture with argon for 30 min. The PdCl ₂ (dppf)-DCM adduct (0.266 g, 0.326 mmol) was added and the mixture was degassed for 20 min. The reaction mixture was heated to 100° C. for 18 hours. The reaction mixture was cooled to room temperature and filtered through a pad of celite and washed with 10% methanol in DCM, then concentrated under reduced pressure. The crude product was dissolved in DCM (50 ml), pre-absorbed onto florisil (50 g) and purified by silica gel chromatography (250 g, 0-100% ethyl acetate in petroleum ether) to give the title compound. LCMS (method L): retention time=2.45, [M+H] ⁺ =354.

Intermediate 24

(S)-(5-bromo-2-methoxyphenyl)(3-hydroxypyrrolidin-1-yl)methanone

DIPEA (1.134 ml, 6.49 mmol) was added to a solution of 5-bromo-2-methoxybenzoic acid (500 mg, 2.164 mmol) and HATU (987 mg, 2.6 mmol) in 2-methyltetrahydrofuran (10 ml) and the reaction mixture was left to stir at room temperature. temp, for 30 min under nitrogen. Then (S)-pyrrolidin-3-ol (226 mg, 2.6 mmol) was added and the reaction mixture was left to stir at room temperature overnight with the addition of dimethylformamide (5 ml). The reaction mixture was concentrated under reduced pressure, diluted with saturated aqueous sodium bicarbonate (50 ml) and 5% lithium chloride solution (50 ml) and extracted with ethyl acetate (2×100 ml). The combined organic layers were passed through a hydrophobic sintered filter, concentrated under reduced pressure, and then dried under nitrogen flow overnight. The residue was taken up in DCM and purified by chromatography on silica gel (80 g) eluting with ethyl acetate:ethanol (3:1, v/v containing 1% triethylamine) in ethyl acetate (0%, 2 column volumes; 0-100 %, 12 column volumes), resulting in the title compound. LCMS (method F): retention time=0.73, [M+H] ⁺ =300.

Intermediate 25

(S)-(3-hydroxypyrrolidin-1-yl)(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone

(S)-(5-bromo-2-methoxyphenyl)(3-hydroxypyrrolidin-1-yl)methanone (intermediate 24, 750 mg, 2.249 mmol), 4,4,4,4',5,5,5',5'-octamethyl-2,2,-bi(1,3,2-dioxaborolane) (628 mg, 2.474 mmol), potassium acetate (662 mg, 6.75 mmol), PdCl ₂ (dppf) (165 mg, 0.225 mmol ) and 1,4-dioxane (6 ml) were combined and placed under nitrogen by evacuation and refilling. The mixture was heated in a microwave reactor at 100° C. for 1 hour. The solvent was removed from the reaction mixture under reduced pressure. The residue was suspended in ethyl acetate (50 ml) and water (30 ml) and filtered through celite, washing with additional ethyl acetate (50 ml). The solution was diluted with brine (20 ml) and the aqueous layer was washed with additional ethyl acetate (100 ml), the combined organic layers were passed through a hydrophobic sintered filter and the solvent was removed under reduced pressure to give the title compound. LCMS (method J): retention time=0.85, [M+H] ⁺ =348.

Intermediate 26

tert-Butyl (5-(1-hydroxyethyl)-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H-imidazol-2-yl)benzyl) carbamate

tert-Butyl(5-formyl-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H-imidazol-2-yl)benzyl)carbamate (intermediate) 15, 465 mg, 0.812 mmol) was dissolved in THF (10 ml) and cooled to -78°C. Methylmagnesium bromide (1 M in dibutyl ether) (0.85 mL, 0.85 mmol) was added dropwise over 2 min. After 1 h 45 min, methylmagnesium bromide (1 M in dibutyl ether) (0.203 mL, 0.203 mmol) was added and the mixture was stirred at -78°C for another 3 h. Saturated aqueous ammonium chloride (5 mL) and water (10 mL) were added. ), and the mixture was stirred for 10 min, warming to room temp. DCM (20 ml) was added, the mixture was transferred and the organic layer was collected. The aqueous layer was then washed with additional dichloromethane (2 x 20 ml), passed the combined organic layers through a hydrophobic sintered filter and evaporated to dryness. The residue was wet-loaded again in DCM (1.5 ml) onto a silica gel column and eluted with 50-60% ethyl acetate in cyclohexane (30 column volumes) followed by 60-85% ethyl acetate in cyclohexane (10 column volumes). Product-containing fractions were combined and evaporated to dryness, then triturated with diethyl ether to give the title compound. LCMS (method F): retention time=1.2, [M+H] ⁺ =589.

Intermediate 27

5-bromo-N-(3-hydroxypropyl)-2-methoxy-N-methylbenzamide

HATU (458.3 mg, 1.205 mmol) and DIPEA (0.655 ml, 3.75 mmol) were added to a stirred solution of 5-bromo-2-methoxybenzoic acid (288.9 mg, 1.250 mmol) in THF (10 ml) at room temp. After 10 minutes, 3-(methylamino)propan-1-ol (0.146 ml, 1.500 mmol) was added. After 2 h, the solvent was removed under vacuum and the resulting residue was diluted with ethyl acetate (10 ml) and water (10 ml). The separated aqueous phase was further extracted with ethyl acetate (2×10 ml). The combined organic layers were passed through a hydrophobic sintered filter and the solvent removed in vacuo to give the title compound. LCMS (method F): retention time=0.78, [M+H] ⁺ =303.

Intermediate 28

N-(3-hydroxypropyl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (1.114 g, 4.39 mmol), potassium acetate (1.077 g , 10.97 mmol), PdCl ₂ adduct (dppf). DCM (0.299 g, 0.366 mmol) and 1,4-dioxane (7 mL) were combined and placed under nitrogen (3 vacuum/nitrogen cycles). A solution of 5-bromo-N-(3-hydroxypropyl)-2-methoxy-N-methylbenzamide (intermediate 27, 1.7 g, 3.66 mmol) in 1,4-dioxane (7 ml) was added and the mixture was treated with 3 vacuum/nitrogen cycles . The reaction mixture was then heated at 100° C. in a microwave reactor for 90 minutes. After cooling, the mixture was filtered through celite, washed with dichloromethane (50 ml) and the solvent was removed under vacuum. The resulting residue was dissolved in ethyl acetate (60 ml), water (60 ml) and brine (20 ml). The separated aqueous layer was re-extracted with ethyl acetate (2×30 ml). The combined organic layers were passed through a hydrophobic sintered filter and the solvent was removed under vacuum. The crude product was purified by silica gel column chromatography eluting with 50-100% ethyl acetate in cyclohexane to give the title compound. LCMS (method F): retention time=0.89, [M+H] ⁺ =350.

Intermediate 29

5-bromo-N-ethyl-N-(2-hydroxyethyl)-2-methoxybenzamide

DIPEA (1.2 ml, 6.87 mmol) HATU (980 mg, 2.58 mmol) was added to a stirred solution of 5-bromo-2-methoxybenzoic acid (500 mg, 2.164 mmol) and the reaction mixture was left to stir overnight. The mixture was partitioned between ethyl acetate (50 ml) and saturated aqueous sodium bicarbonate (50 ml). The aqueous phase was further extracted with ethyl acetate (50 ml). The combined organic phase was washed with saturated aqueous sodium bicarbonate (50 ml) and the solvent was removed under reduced pressure. The residue was purified by silica gel chromatography eluting with 0-100% ethyl acetate:ethanol (3:1 containing 1% triethylamine) in cyclohexane to give the title compound. LCMS (method F): retention time=0.8, [M+H] ⁺ =302.

Intermediate 30

N-ethyl-N-(2-hydroxyethyl)-2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

5-bromo-N-ethyl-N-(2-hydroxyethyl)-2-methoxybenzamide (intermediate 29, 1.2 g, 3.97 mmol), 4,4,4',4',5,5,5',5' -octamethyl-2,2,-bi(1,3,2-dioxaborolane) (1.18 g, 4.65 mmol), PdCl ₂ (dppf) (0.301 g, 0.411 mmol), potassium acetate (1.17 g, 11.92 mmol) and 1 ,4-dioxane (8 ml) was combined and placed under nitrogen by evacuation and refilling. The mixture was heated at 100°C in a microwave reactor for 1.5 h. After cooling, the reaction mixture was filtered through celite, washed with methanol (250 ml) and the solvent was removed under reduced pressure. The residue was redissolved in DCM (30 ml) and partitioned with water (30 ml). The aqueous layer was re-extracted with dichloromethane (3 x 30 mL), the combined organic extracts were passed through a hydrophobic sintered filter and evaporated to dryness to give the title compound. LCMS (method F): rt 0.92, [M+H] ⁺ =350.

Intermediate 31

5-bromo-2-chloro-N-ethyl-N-(2-hydroxyethyl)benzamide

DIPEA (1.1 ml, 6.30 mmol) and HATU (1.05 g, 2.76 mmol) were added to a stirred solution of 5-bromo-2-chlorobenzoic acid (509 mg, 2.162 mmol) in 2-methyltetrahydrofuran (10 ml) and the reaction mixture was left to stir at room temp for 15 min. 2-(ethylamino)ethan-1-ol (0.25 ml, 2.56 mmol) was added and the reaction mixture was stirred over the weekend. The mixture was partitioned between ethyl acetate (50 ml) and saturated aqueous sodium bicarbonate (50 ml). The aqueous phase was then washed with ethyl acetate (50 ml) and the combined organic phases were washed with saturated aqueous sodium bicarbonate (50 ml) and the solvent was removed under reduced pressure. The crude product was purified by silica gel column chromatography eluting with 20-100% ethyl acetate in cyclohexane. Fractions containing the product were combined and evaporated to dryness. The residue was then separated by reverse phase column chromatography eluting with 20-60% acetonitrile (containing 0.01% ammonia) in 10 mM ammonium bicarbonate in water adjusted to pH 10 with ammonia to give the title compound. LCMS (method E): retention time=0.85, [M+H] ⁺ =306.

Intermediate 32

2-chloro-N-ethyl-N-(2-hydroxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

4,4,4',4',5,5,5',5'-octamethyl-2,2,-bi(1,3,2-dioxaborolane) (462 mg, 1.819 mmol), 5-bromo-2 -chloro-N-ethyl-N-(2-hydroxyethyl)benzamide (intermediate 31, 453 mg, 1.478 mmol), PdCl ₂ (dppf) (111 mg, 0.152 mmol) and potassium acetate (445 mg, 4.53 mmol) were combined in 1,4-dioxane (4 ml). The mixture was degassed and heated at 100°C in a microwave reactor for 1 hour, three times. After cooling, the mixture was filtered through celite, washed with methanol (100 ml) and the solvent was removed under vacuum. The residue was partitioned between DCM (20 ml) and water (10 ml). Then the separated aqueous layer was washed with dichloromethane (2×10 ml). The combined organic layers were passed through a hydrophobic sintered filter and the solvent removed in vacuo to give the title compound. LCMS (method D): retention time=1.05, [M+H] ⁺ =354.

Intermediate 33

5-bromo-2-methoxy-N-methyl-N-(tetrahydro-2H-pyran-4-yl)benzamide

HATU (1.391 g, 3.66 mmol) and DIPEA (1.72 ml, 9.85 mmol) were added to a stirred suspension of 5-bromo-2-methoxybenzoic acid (0.752 g, 3.25 mmol) in THF (10 ml) and stirred for 20 min. N-methyltetrahydro-2H-pyran-4-amine (0.382 g, 3.32 mmol) was added and the reaction mixture was left to stir for 18 h. The solvent was removed under vacuum. The reaction mixture was partitioned between DCM (50 ml) and water (50 ml), then the aqueous phase was extracted with dichloromethane (2×20 ml) and the organic phase was filtered through a hydrophobic sintered filter and the solvent was removed under vacuum. The residue was dissolved in a minimum amount of DCM and purified by chromatography on silica gel (120 g) eluting with 0-100% ethyl acetate:ethanol (3:1, v/v) in cyclohexane to give the title compound. LCMS (method F): retention time=0.89, [M+H] ⁺ =328.

Intermediate 34

2-methoxy-N-methyl-N-(tetrahydro-2H-pyran-4-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

5-bromo-2-methoxy-N-methyl-N-(tetrahydro-2H-pyran-4-yl)benzamide (intermediate 33, 746 mg, 2.273 mmol), 4,4,4',4',5, 5,5,,5,-octamethyl-2,2,-bi(1,3,2-dioxaborolane) (692 mg, 2.73 mmol), PdCl ₂ (dppf). DCM (194 mg, 0.238 mmol), potassium acetate (664 mg, 6.77 mmol) and 1,4-dioxane (10 ml) were combined and heated in a microwave reactor at 100°C for 1 h. The reaction mixture was filtered through celite and the solvent was removed under vacuum. The residue was partitioned between water (25 ml) and DCM (25 ml), then the aqueous phase was extracted with dichloromethane (2×15 ml) and the organic phase was filtered through a hydrophobic sintered filter, then evaporated to dryness, whereby the title compound was obtained. LCMS (method F): retention time=1.01, [M+H] ⁺ =376.

Intermediate 35

5-bromo-N-(3-hydroxypropyl)-N,2-dimethylbenzamide

5-bromo-2-methylbenzoic acid (504 mg, 2.344 mmol), HATU (1053 mg, 2.77 mmol) and DIPEA (0.5 ml, 2.86 mmol) in DMF (4 ml) was stirred at room temperature for 30 min, after to which 3-(methylamino)propan-1-ol (0.275 ml, 2.83 mmol) was added. After 3 hours the mixture was partitioned between ethyl acetate (20 ml) and water (15 ml). The separated aqueous layer was further extracted with ethyl acetate (15 ml). The combined organic layers were washed with 5% aqueous lithium chloride solution (2×10 ml), water (10 ml) and brine (10 ml). The organic layer was passed through a hydrophobic sintered filter and the solvent was removed under vacuum. The crude product was purified by silica gel column chromatography eluting with 50-100% ethyl acetate in cyclohexane to give the title compound. LCMS (method F): retention time=0.80, [M+H] ⁺ =286.

Intermediate 36

N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

Potassium acetate (636 mg, 6.48 mmol), 5-bromo-N-(3-hydroxypropyl)-N,2-dimethylbenzamide (Intermediate 35, 685 mg, 2.154 mmol), 4,4,4',4',5 ,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (663 mg, 2.61 mmol) and PdCl ₂ (dppf) (131 mg, 0.179 mmol) were combined into 1, 4-dioxane (5 ml). The mixture was placed under nitrogen by five vacuum/nitrogen cycles and heated in a microwave at 100° C. for 1 hour. After cooling, the solvent was removed under vacuum and the residue was partitioned between DCM (30 ml) and water (20 ml). The separated aqueous layer was then washed with dichloromethane (2 x 20 mL) and the combined organic layers were passed through a hydrophobic sintered filter and the solvent removed in vacuo to give the title compound. LCMS (method J): retention time=0.97, [M+H] ⁺ =334.

Intermediate 37

(S)-(5-bromo-2-chlorophenyl)(3-hydroxypyrrolidin-1-yl)methanone

Prepared analogously to Intermediate 33 using 5-bromo-2-chlorobenzoic acid (480 mg, 2.039 mmol) and (S)-pyrrolidin-3-ol (178 mg, 2.039 mmol) to give the title compound. LCMS (method F): retention time=0.73, [M+H] ⁺ =303.

Intermediate 38

(S)-(2-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)(3-hydroxypyrrole idin-1-yl)methanone

Prepared analogously to Intermediate 32 using (S)-(5-bromo-2-chlorophenyl)(3-hydroxypyrrolidin-1-yl)methanone (Intermediate 37) to give the title compound. LCMS (method F): retention time=0.72, [M+H] ⁺ =352.

Intermediate 39

(S)-5-bromo-N-1(-hydroxypropan-2-yl)-2-methoxy-N-methylbenzamide

Prepared analogously to intermediate 35 using 5-bromo-2-methoxybenzoic acid (0.747 g, 3.23 mmol) and (S)-2-(methylamino)propan-1-ol (0.321 g, 3.6 mmol) to give the title connection header. LCMS (method F): retention time=0.77, [M+H] ⁺ =302.

Intermediate 40

(S)-N-(1-hydroxypropan-2-yl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

Prepared analogously to Intermediate 32 using (S)-5-bromo-N-(1-hydroxypropan-2-yl)-2-methoxy-N-methylbenzamide (Intermediate 39) to give the title compound. LCMS (method F): retention time=0.89, [M+H] ⁺ =350.

Intermediate 41

Ethyl 3-bromo-7-chloro-2-methylpyrazolo[1,5-а]pyrimidine-5-carboxylate

Prepared similarly to Intermediate 6 using BSI to give the title compound. LCMS (method J): retention time=1.11, [M+H] ⁺ =320.

Intermediate 42

Ethyl 3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidine-5-carboxylate

Ethyl 3-bromo-7-chloro-2-methylpyrazolo[1,5-a]pyrimidine-5-carboxylate (Intermediate 41, 685 mg, 2.15 mmol), (3-(1-methyl-1H-imidazole-2 -yl)phenyl)methanamine (intermediate 3, 443 mg, 2.365 mmol) and DIPEA (0.751 ml, 4.3 mmol) were stirred in DMSO (5 ml) at 80°C under nitrogen for 3.5 h. After cooling, the mixture was poured into ice water and collected the precipitate by filtration. The filter cake was dissolved in DCM, dried under reduced pressure and triturated with diethyl ether to give the title compound. LCMS (method F): retention time=1.06, [M+H] ⁺ =469.

Alternative route for intermediate 42

Ethyl 3-bromo-7-chloro-2-methylpyrazolo[1,5-a]pyrimidine-5-carboxylate (Intermediate 41, 50 g, 157 mmol), (3-(1-methyl-1H-imidazole-2 -yl)phenyl)methanamine (intermediate 3, 35.3 g, 188 mmol) and DIPEA (54.8 ml, 314 mmol) were stirred in DMSO (500 ml) at 80°C for 3 h. After cooling, the reaction mixture was poured into ice water and collected the precipitate by filtration. The resulting solid was triturated with diethyl ether (500 ml), filtered and dried to give the title compound. LCMS (method C): retention time=1.44, [M+H] ⁺ =469.

Intermediate 43

2-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)propan- 2-ol

Ethyl 3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidine-5-carboxylate (intermediate 42, 1.76 g, 3.75 mmol) was stirred in THF (35 ml) at 0°C under nitrogen atmosphere. Methylmagnesium chloride (3 M in THF) (5.6 ml, 16.8 mmol) was added over 10 minutes and the mixture was stirred at 0°C for 30 minutes. The mixture was warmed to room temperature and stirred overnight. The reaction mixture was quenched with saturated aqueous ammonium chloride (5 ml), then separated with dichloromethane (100 ml) and saturated aqueous ammonium chloride (100 ml). The organic phase was collected and the aqueous was washed with dichloromethane (2×25 ml). The combined organic layers were passed through a hydrophobic sintered filter and evaporated to dryness. The residue was stirred in THF (35 ml) under nitrogen at 0°C. Methylmagnesium chloride (3 M in THF) (3.75 ml, 11.25 mmol) was added over 10 minutes and the mixture was stirred at 0°C for 40 minutes, after which it was stirred at room temperature for 20 minutes. The reaction mixture was quenched with saturated aqueous ammonium chloride (5 ml), then separated with dichloromethane (100 ml) and saturated aqueous ammonium chloride (100 ml). The organic phase was collected and the aqueous was washed with dichloromethane (2×25 ml). The combined organic layers were passed through a hydrophobic sintered filter, concentrated under reduced pressure, and dried for 4 days in a high vacuum unit to give the title compound. LCMS (method J): retention time=0.65, [M+H] ⁺ =455.

Alternative route for intermediate 43

Ethyl 3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidine-5-carboxylate (intermediate 42 , 81 g, 173 mmol) was stirred in DCM (300 ml) and THF (100 ml) at 5° C. under nitrogen. Methylmagnesium chloride (3 M in THF) (230 ml, 690 mmol) was added dropwise over 45 minutes keeping the internal temperature below 15°C and stirred for another 10 minutes. The mixture was quenched with saturated aqueous ammonium chloride (400 ml) and partitioned between DCM (2 L) and water (2 L). The organic phase was dried with magnesium sulfate and concentrated under reduced pressure. The resulting residue was dissolved in DCM (500 ml) and THF (250 ml) and cooled to 0° C. under nitrogen. Methylmagnesium chloride (3 M in THF) (173 ml, 518 mmol) was added dropwise over 1 h while maintaining the internal temperature below 10°C. After the addition, the mixture was stirred at 5° C. for 10 minutes. The mixture was quenched with saturated aqueous ammonium chloride (400 ml) and partitioned between DCM (2 L) and water (2 L). The organic phase was dried with magnesium sulfate and concentrated under reduced pressure to give the title compound. LCMS (method J): retention time=0.60, [M+H] ⁺ =455.

Intermediate 44

(S)-5-bromo-N-(1-hydroxypropan-2-yl)-N,2-dimethylbenzamide

Prepared analogously to Intermediate 33 using 5-bromo-2-methylbenzoic acid (800 mg, 3.72 mmol) and (S)-2-(methylamino)propan-1-ol (332 mg, 3.72 mmol) to give the title connection header. LCMS (method F): retention time=0.82, [M+H] ⁺ =286.

Intermediate 45

(S)-N-(1-hydroxypropan-2-yl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

Prepared analogously to Intermediate 32 using (S)-5-bromo-N-(1-hydroxypropan-2-yl)-N,2-dimethylbenzamide (Intermediate 44) to give the title compound. LCMS (method F): retention time=0.92, [M+H] ⁺ =334.

Intermediate 46

5-bromo-2-methoxy-N,N-dimethylbenzamide

Prepared analogously to intermediate 33 using 5-bromo-2-methoxybenzoic acid (483 mg, 2.091 mmol) and dimethylamine (2 M in THF) (1.254 ml, 2.509 mmol) to give the title compound. LCMS (method F): retention time=0.85, [M+H] ⁺ =258.

Intermediate 47

2-methoxy-N,N-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

Prepared analogously to Intermediate 32 using 5-bromo-2-methoxy-N,N-dimethylbenzamide (Intermediate 46) to give the title compound. LCMS (method F): retention time=1.01, [M+H] ⁺ =306.

Intermediate 48

(R)-(5-bromo-2-chlorophenyl)(2-(hydroxymethyl)pyrrolidin-1-yl)methanone

Prepared analogously to Intermediate 33 using 5-bromo-2-chlorobenzoic acid (503 mg, 2.136 mmol) and (R)-pyrrolidin-2-ylmethanol to give the title compound. LCMS (method F): retention time=0.89, [M+H] ⁺ =320.

Intermediate 49

(R)-(2-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)(2-(hydroxymethyl)pyrrolidin-1-yl)methanone

Prepared analogously to Intermediate 32 using (R)-(5-bromo-2-chlorophenyl)(2-(hydroxymethyl)pyrrolidin-1-yl)methanone (Intermediate 48) to give the title compound. LCMS (method F): retention time=0.94, [M+H] ⁺ =366.

Intermediate 50

(R)-(5-bromo-2-methylPhenyl)(2-(hydroxymethyl)pyrrolidin-1-yl)methanone

5-bromo-2-methylbenzoic acid (534 mg, 2.483 mmol), HATU (1074 mg, 2.82 mmol) and DIPEA (0.520 ml, 2.98 mmol) in DMF (1 ml) and THF (5 ml) were stirred at room temperature, for 30 min, then (R)-pyrrolidin-2-ylmethanol (0.3 ml, 3.04 mmol) was added in one portion. The reaction mixture was stirred for 18 hours. The solvent was removed under vacuum. The residue was taken up in ethyl acetate (40 ml) and partitioned with water (20 ml). The organic layer was separated and washed with 5% aqueous lithium chloride solution (2×10 ml), water (10 ml) and brine (10 ml). The organic layer was then passed through a hydrophobic sintered filter and the solvent removed under vacuum. The residue was dissolved in a minimum amount of DCM and loaded onto a silica gel (80 g) column and eluted with 12 column volumes of 0-100% ethyl acetate:ethanol (3:1, v/v containing 1% triethylamine) in cyclohexane, resulting in which gave the title compound. LCMS (method F): retention time=0.87, [M+H] ⁺ =298.

Intermediate 51

(R)-(2-(hydroxymethyl)pyrrolidin-1-yl)(2-methyl-5-(4,4,5,,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone

Prepared analogously to Intermediate 36 using (R)-(5-bromo-2-methylphenyl)(2-(hydroxymethyl)pyrrolidin-1-yl)methanone (Intermediate 50) to give the title compound. LCMS (method F): retention time=1.02, [M+H] ⁺ =346.

Intermediate 52

5-bromo-N-(2-hydroxyethyl)-2-methoxy-N-methylbenzamide

Prepared analogously to Intermediate 33 using 5-bromo-2-methoxybenzoic acid (400 mg, 1.731 mmol) and 2-(methylamino)ethan-1-ol (0.167 mL, 2.078 mmol) to give the title compound. LCMS (method F): retention time=0.72, [M+H] ⁺ =288.

Intermediate 53

N-(2-hydroxyethyl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

Prepared analogously to Intermediate 32 using 5-bromo-N-(2-hydroxyethyl)-2-methoxy-N-methylbenzamide (Intermediate 52) to give the title compound. LCMS (method F): retention time=0.85, [M+H] ⁺ =336.

Intermediate 54

(5-bromo-2-methoxyphenyl)(3-hydroxy-3-methylpyrrolidin-1-yl)methanone

Prepared analogously to Intermediate 33 using 5-bromo-2-methoxybenzoic acid (503 mg, 2.177 mmol) and 3-methylpyrrolidin-3-ol (244 mg, 2.412 mmol) to give the title compound. LCMS (method F): retention time=0.76, [M+H] ⁺ =314.

Intermediate 55

(3-hydroxy-3-methylpyrrolidin-1-yl)(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone

Prepared analogously to intermediate 32 using (5-bromo-2-methoxyphenyl)(3-hydroxy-3-methylpyrrolidin-1-yl)methanone (intermediate

compound 54) to give the title compound. LCMS (method F): retention time=0.91, [M+H] ⁺ =362.

Intermediate 56

(5-bromo-2-methoxyphenyl)(1,4-oxazepan-4-yl) methanone

Prepared analogously to Intermediate 33 using 5-bromo-2-methoxybenzoic acid (528 mg, 2.285 mmol) and 1,4-oxazepane (272 mg, 2.69 mmol) to give the title compound. LCMS (method F): retention time=0.87, [M+H] ⁺ =316.

Intermediate 57

(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)(1,4-oxazepan-4-yl) methanone

Prepared analogously to Intermediate 32 using (5-bromo-2-methoxyphenyl)(1,4-oxazepan-4-yl)methanone (Intermediate 56) to give the title compound. LCMS (method F): retention time=1.0, [M+H] ⁺ =362.

Intermediate 58

(S)-(5-bromo-2-methylPhenyl)(3-hydroxypyrrolidin-1-yl)methanone

Prepared analogously to Intermediate 29 using 5-bromo-2-methylbenzoic acid (493 mg, 2.293 mmol) and (S)-pyrrolidin-3-ol (0.23 mL, 2.77 mmol) to give the title compound. LCMS (method F): retention time=0.75, [M+H] ⁺ =284.

Intermediate 59

(S)-(3-hydroxypyrrolidin-1-yl)(2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone

Prepared analogously to Intermediate 32 using (S)-(5-bromo-2-methylphenyl)(3-hydroxypyrrolidin-1-yl)methanone (Intermediate 58) to give the title compound. LCMS (method F): retention time=0.87, [M+H] ⁺ =332.

Intermediate 60

(S)-(5-bromo-2-methoxyphenyl)(3-methylmorpholino)methanone

Prepared analogously to Intermediate 50 using 5-bromo-2-methoxybenzoic acid (505 mg, 2.186 mmol) and (S)-3-methylmorpholine (251 mg, 2.481 mmol) to give the title compound. LCMS (method F): retention time=0.92, [M+H] ⁺ =314.

Intermediate 61

(S)-(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)(3-methylmorpholino)methanone

Prepared analogously to Intermediate 32 using (S)-(5-bromo-2-methoxyphenyl)(3-methylmorpholino)methanone (Intermediate 60) to give the title compound. LCMS (method F): retention time=1.04, [M+H] ⁺ =362.

Intermediate 62

(5-bromo-2-methylPhenyl)(3-hydroxy-3-methylpyrrolidin-1-yl)methanone

Prepared analogously to Intermediate 33 using 5-bromo-2-methylbenzoic acid (394 mg, 1.832 mmol) and 3-methylpyrrolidin-3-ol (222 mg, 2.199 mmol) to give the title compound. LCMS (method F): retention time=0.81, [M+H] ⁺ =298.

Intermediate 63

(3-hydroxy-3-methylpyrrolidin-1-yl)(2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone

Prepared analogously to Intermediate 32 using (5-bromo-2-methylphenyl)(3-hydroxy-3-methylpyrrolidin-1-yl)methanone (Intermediate 62) to give the title compound. LCMS (method F): retention time=0.91, [M+H] ⁺ =346.

Intermediate 64

Ethyl 7-((3-(1H-pyrazol-1-yl)benzyl)amino)-3-bromo-2-methylpyrazolox[1,5-a]pyrimidine-5-carboxylate

Prepared analogously to Intermediate 42 using (3-(1H-pyrazol-1-yl)phenyl)methanamine (available from Sigma-Aldrich Inc.) to give the title compound. LCMS (method F): retention time=1.21, [M+H] ⁺ =455.

Intermediate 65

Ethyl 7-((3-(1H-pyrazol-1-yl)benzyl)amino)-3-bromo-2-methylpyrazoloI,5-a1pyrimidine-5-carboxylate

Prepared analogously to intermediate 43 using ethyl 7-((3-(1H-pyrazol-1-yl)benzyl)amino)-3-bromo-2-methylpyrazolo[1,5-a]pyrimidine-5-carboxylate (intermediate compound 64) to give the title compound. LCMS (method F): retention time=1.20, [M+H] ⁺ =441.

Intermediate 66

(R)-(5-bromo-2-methoxyphenyl)(2-(hydroxymethyl)pyrrolidin-1-yl)methanone

Prepared analogously to Intermediate 35 using 5-bromo-2-methoxybenzoic acid and (R)-pyrrolidin-2-ylmethanol to give the title compound. LCMS (method F): retention time=0.83, [M+H] ⁺ =314.

Intermediate 67

(R)-(2-(Hydroxymethyl)pyrrolidin-1-yl)(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone

Prepared analogously to Intermediate 36 using (R)-(5-bromo-2-methoxyphenyl)(2-(hydroxymethyl)pyrrolidin-1-yl)methanone (Intermediate 66) to give the title compound. LCMS (method F): retention time=1.02, [M+H] ⁺ =362.

Intermediate 68

5-bromo-2-chloro-N-(2-hydroxyethyl)-N-methylbenzamide

Prepared analogously to Intermediate 29 using 5-bromo-2-chlorobenzoic acid and 2-(methylamino)ethan-1-ol in THF to give the title compound. LCMS (method F): retention time=0.77, [M+H] ⁺ =294.

Intermediate 69

2-chloro-N-(2-hydroxyethyl)-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

Prepared similarly to Intermediate 36 using Intermediate 68 to give the title compound. LCMS (method F): retention time=0.75, [M+H] ⁺ =340.

Intermediate 70

5-bromo-N-(2-hydroxyethyl)-N,2-dimethylbenzamide

Prepared analogously to Intermediate 29 using 5-bromo-2-methylbenzoic acid and 2-(methylamino)ethan-1-ol to give the title compound. LCMS (method F): retention time=0.76, [M+H] ⁺ =274.

Intermediate 71

N-(2-hydroxyethyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

Prepared similarly to Intermediate 36 using Intermediate 70 to give the title compound. LCMS (method F): retention time=0.85, [M+H] ⁺ =320.

Intermediate 72

5-bromo-N-(2-hydroxyethyl)-N,2-dimethylbenzamide

Prepared analogously to Intermediate 29 using 5-bromo-2-methylbenzoic acid and 2-(ethylamino)ethan-1-ol in THF to give the title compound. LCMS (method F): retention time=0.84, [M+H] ⁺ =286.

Intermediate 73 N-Ethyl-N-(2-hydroxyethyl)-2-methyl-5-(4,4,5,5-tetramethyl1,3,2-dioxaborolan-2-yl)benzamide

Prepared similarly to Intermediate 36 using Intermediate 72 to give the title compound. LCMS (method F): retention time=0.95, [M+H] ⁺ =334.

Intermediate 74

Ethyl 2-(2-methyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)acetate

To a stirred solution of 5-methyl-1H-pyrazol-3-amine (25 g, 257 mmol) in 1,4-dioxane (150 mL) was added diethyl 3-oxo-pentanedioate (56.8 mL, 309 mmol) followed by acetic acid (7.37 ml, 129 mmol) at room temp. The reaction mixture was stirred at 100° C. for 6 hours. The mixture was allowed to cool to room temp. The solids were collected by filtration, washed with ethyl ether and dried to give the title compound. LCMS (method H): retention time=1.55, [M+H] ⁺ =236.

Intermediate 75

Ethyl 2-(7-chloro-2-methylpyrazolo[1,5-a]pyrimidin-5-yl)acetate

To a stirred solution of ethyl 2-(2-methyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)acetate (intermediate 74, 22.5 g, 96 mmol) in acetonitrile (250 ml), DIPEA (33.4 ml, 191 mmol), N-methylmorpholine (0.105 ml, 0.956 mmol), and POCl ₃ (17.83 ml, 191 mmol) were added at 0°С. The reaction mixture was stirred at room temperature for 24 hours. After completion of the reaction, the mixture was concentrated and then poured into a mixture of water and people. The mixture was neutralized with sodium bicarbonate and extracted with ethyl acetate (2×300 ml). The combined organic layers were washed with brine, dried using sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (0-20% ethyl acetate in hexanes) to give the title compound. LCMS (method H): retention time=2.05, [M+H] ⁺ =254.

Intermediate 76

Ethyl 2-(7-chloro-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-5-yl)acetate

To a stirred solution of ethyl 2-(7-chloro-2-methylpyrazolo[1,5-a]pyrimidin-5-yl)acetate (intermediate 75, 8 g, 31.5 mmol) in DMF (160 ml) was added N-iodosuccinimide (7.09 g, 31.5 mmol) at 0°С. The resulting reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was poured onto ice-cold water and the precipitated solid was collected by filtration. The residue was purified by silica gel chromatography (0-10% ethyl acetate in hexanes) to give the title compound. LCMS (method H): retention time=2.43, [M+H] ⁺ =380.

Intermediate 77

Ethyl 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)acetate

A mixture of ethyl 2-(7-chloro-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-5-yl)acetate (intermediate 76, 6 g, 15.81 mmol), (3-(1-methyl- 1H-imidazol-2-yl)phenyl)methanamine (intermediate 3, 3.55 g, 18.97 mmol) and DIPEA (5.52 ml, 31.6 mmol) in DMSO (50 ml) were stirred and heated at 60°C for 2 h. After cooling, the reaction mixture was poured into ice-cold water (300 ml), and the precipitated solid was collected by filtration, then dried under reduced pressure, whereby the title compound was obtained. LCMS (method H): retention time=1.74, [M+H] ⁺ =531.

Intermediate 78

Methyl 2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate

Solution/suspension of methyl 5-iodo-2-methoxybenzoate (20 g, 68.5 mmol), bis (pinacolato) diboron (17.39 g, 68.5 mmol), potassium acetate (20.16 g, 205 mmol) and bis (triphenylphosphine) palladium (II ) chloride (3.85 g, 5.48 mmol) in 1,4-dioxane (200 mL) was heated to 100°С under nitrogen atmosphere. The reaction mixture was stirred at 100° C. for 20 h and then allowed to cool. The reaction mixture was filtered through a pad of celite. The Celite layer was washed with ethyl acetate (250 ml). 1 M hydrochloric acid (250 ml) was added to the filtrate. The organic phase was washed with brine (100 ml) and then dried with magnesium sulfate. The solvent was removed under vacuum. This was dissolved in DCM and applied to a silica gel cartridge (750 g) and eluted with a gradient of 0-50% ethyl acetate in cyclohexane using 9 column volumes. The desired fractions were combined and evaporated in vacuo to give the title compound. LCMS (method J): retention time=1.16, [M+H] ⁺ =293.

Intermediate 79 and Intermediate 80

Ethyl 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl) propanoate and ethyl 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5- yl)-2-methylpropanoate

In a dried vessel, ethyl 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5 -yl)acetate (intermediate 77, 400 mg, 0.754 mmol) was stirred in THF (4.0 ml) under nitrogen at 0°C. Methyl iodide (0.05 ml, 0.8 mmol) was added and the mixture was stirred at 0°C for 1 min, after which LiHMDS (1 M in THF) (1.6 ml, 1.6 mmol) was added. The reaction mixture was stirred at 0°C for 5 min, then allowed to warm to room temperature. After 15 minutes the reaction mixture was diluted with saturated aq. NH ₄ Cl (2 ml) and stirred for 5 min. The mixture was transferred to a separating funnel with dichloromethane (20 ml) and water (20 ml). The phases were separated and the organic layer was collected. The aqueous layer was then washed with additional dichloromethane (2×10 ml) and the combined organic layers were washed with brine (20 ml), filtered through a hydrophobic sintered filter and evaporated to dryness. The crude product was wet loaded from DCM (1 ml) onto a 28 g KP-NH column and eluted with 10-50% ethyl acetate:ethanol (3:1 containing 1% triethylamine) in cyclohexane. The sample was then purified by reverse phase chromatography eluting with 40-90% acetonitrile in pH10 buffered ammonium carbonate water. Fractions containing each product were combined separately, concentrated under reduced pressure, and dried in a vacuum oven over the weekend to give the title compounds of interest. Intermediate 79: LCMS (method F): retention time=1.16, [M+H] ⁺ =545. Intermediate 80: LCMS (method F): retention time=1.27, [M+H] ⁺ =559.

Intermediate 81

2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)propan- 1-ol

Ethyl-2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl) propanoate (intermediate 79, 87 mg, 0.160 mmol) was stirred in THF (1.5 mL) at 0°С under nitrogen atmosphere. DIBAL-H (1 M in THF) (0.5 ml, 0.5 mmol) was added dropwise and the mixture was stirred for 45 min. The mixture was neutralized with an aqueous 1 M Rochelle's salt solution and stirred vigorously for 30 minutes. The mixture was separated with dichloromethane (10 ml) and water (10 ml). The separated aqueous phase was washed with additional dichloromethane (2 x 10 mL) and the combined organic phases were passed through a hydrophobic sintered filter and concentrated under reduced pressure to give the title compound. LCMS (method F): retention time=0.99, [M+H] ⁺ =503.

Intermediate 82

2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-а]pyrimidin-5-yl)-2 -methylpropan-1-ol

Prepared analogously to intermediate 81 using ethyl 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidin-5-yl)-2-methylpropanoate (intermediate 80) to give the title compound. LCMS (method F): retention time=1.12, [M+H] ⁺ =517.

Intermediate 83

(5-bromo-2-methoxyphenyl)(2-(hydroxymethyl)pyrrolidin-1-yl)methanone

Prepared analogously to Intermediate 29 using 5-bromo-2-methoxybenzoic acid and pyrrolidin-2-ylmethanol in THF to give the title compound. LCMS (method F): retention time=0.84, [M+H] ⁺ =314.

Intermediate 84

(2-(hydroxymethyl)pyrrolidin-1-yl)(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone

Prepared similarly to Intermediate 36 using Intermediate 83 to give the title compound. LCMS (method F): retention time=0.96, [M+H] ⁺ =362.

Intermediate 85

(R)-5-bromo-N-(1-hydroxypropan-2-yl)-N,2-dimethylbenzamide

Prepared analogously to Intermediate 29 using 5-bromo-2-methoxybenzoic acid and (R)-2-(methylamino)propan-1-ol in THF to give the title compound. LCMS (method F): retention time=0.82, [M+H] ⁺ =286.

Intermediate 86

(R)-N-(1-hydroxypropan-2-yl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

Prepared similarly to Intermediate 36 using Intermediate 85 to give the title compound. LCMS (method F): retention time=0.91, [M+H] ⁺ =334.

Intermediate 87

5-bromo-N-(1-hydroxypropan-2-yl)-N,2-dimethylbenzamide

Prepared analogously to Intermediate 29 using 5-bromo-2-methylbenzoic acid and 2-(methylamino)propan-1-ol in THF to give the title compound. LCMS (method F): retention time=0.82, [M+H] ⁺ =286.

Intermediate 88 N-(1-hydroxypropan-2-yl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

Prepared analogously to Intermediate 36 using 5-bromo-N-(1-hydroxypropan-2-yl)-N,2-dimethylbenzamide (Intermediate 87) to give the title compound. LCMS (method F): retention time=0.91, [M+H] ⁺ =334.

Intermediate 89

(5-bromo-2-methylphenyl)(3-hydroxypyrrolidin-1-yl)methanone

Prepared analogously to Intermediate 29 using 5-bromo-2-methylbenzoic acid and pyrrolidin-3-ol in THF to give the title compound. LCMS (method F): retention time=0.75, [M+H] ⁺ =284.

Intermediate 90

(3-hydroxypyrrolidin1-yl)(2-methyl-5-(4,4,5,5-tetramethyl1,3,2-dioxaborolan-2-yl)phenyl)methanone

Prepared analogously to Intermediate 36 using (5-bromo-2-methylphenyl)(3-hydroxypyrrolidin-1-yl)methanone (Intermediate 89) to give the title compound. LCMS (method F): retention time=0.84, [M+H] ⁺ =332.

Intermediate 91

5-bromo-N-(1-hydroxypropan-2-yl)-2-methoxy-N-methylbenzamide

Prepared analogously to Intermediate 29 using 5-bromo-2-methoxybenzoic acid and 2-(methylamino)propan-1-ol in THF to give the title compound. LCMS (method F): retention time=0.76, [M+H] ⁺ =302.

Intermediate 92

N-(1-hydroxypropan2-yl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

Prepared analogously to Intermediate 36 using 5-bromo-N-(1-hydroxypropan-2-yl)-2-methoxy-N-methylbenzamide (Intermediate 91) to give the title compound. LCMS (method F): retention time=0.89, [M+H] ⁺ =350.

Intermediate 93 and Intermediate 94

tert-Butyl (5-(1-hydroxyethyl)-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H-imidazol-2-yl)benzyl) carbamate, isomer 1 and isomer 2

tert-Butyl (5-(1-hydroxyethyl)-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H-imidazol-2-yl)benzyl) carbamate (intermediate 26, 60 mg, 0.123 mmol) was separated using a Chiralpak AD-H column (30 mm × 250 mm, 5 µm) and a solvent system: 30% ethanol (containing 0.2% isopropylamine)/heptane (containing 0.2% isopropylamine ) at 30 ml/min to give the chiral title compounds, isomer 1: LCMS (method F): retention time=0.97, [M+H] ⁺ =489. Chiral HPLC: retention time 10.73, 99.7%. isomer 2: LCMS (method F): retention time=0.97, [M+H] ⁺ =489. Chiral HPLC: retention time 7.53, 100%.

Intermediate 95

5-bromo-N,2-dimethyl-N-(tetrahydro-2H-pyran-4-yl)benzamide

Prepared analogously to Intermediate 33 using 5-bromo-2-methylbenzoic acid and N-methyltetrahydro-2H-pyran-4-amine in THF to give the title compound. LCMS (method F): retention time=0.96, [M+H] ⁺ =312.

Intermediate 96

N,2-dimethyl-N-(tetrahydro-2H-pyran-4-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

Prepared analogously to Intermediate 32 using 5-bromo-N,2-dimethyl-N-(tetrahydro-2H-pyran-4-yl)benzamide (Intermediate 95) to give the title compound. LCMS (method F): retention time=1.06, [M+H] ⁺ =360.

Intermediate 97

2-methylpyrazolo[1,5-а]pyrimidine-5,7(4H,6H)-dione

5-methyl-1H-pyrazole-3-amine (50 g, 515 mmol) was dissolved in ethanol (500 ml) under nitrogen atmosphere. Sodium ethanolate (334 g, 1030 mmol) in ethanol was added followed by diethyl malonate (86 ml, 566 mmol). The reaction mixture was stirred at 80° C. for 4 hours. The reaction mixture was allowed to cool to room temperature and the precipitated solid was collected by filtration, washed with excess ethanol (1000 ml) and dried to give the title compound. LCMS (method B): retention time=0.48, [M+H] ⁺ =166.

Intermediate 98

5,7-dichloro-2-methylpyrazolo[1,5-а]pyrimidine

2-methylpyrazolo[1,5-a]pyrimidine-5,7(4H,6H)-dione (intermediate 97, 35 g, 212 mmol) was added to phosphorus oxychloride (395 ml, 4239 mmol) and the mixture was heated at 100° C for 24 hours. The mixture was cooled to room temperature and excess phosphorus oxychloride was removed under reduced pressure. The mixture was co-distilled with toluene (2×200 ml). The crude product was purified by silica gel column chromatography (20% ethyl acetate in petroleum ether) to give the title compound. LCMS (method B): retention time=3.21, [M+H] ⁺ =201.

Intermediate 99

5,7-dichloro-3-iodo-2-methylpyrazolo[1,5-a]pyrimidine

To a stirred solution of 5,7-dichloro-2-methylpyrazolo[1,5-a7-pyrimidine (intermediate 98, 15 g, 74.2 mmol) in DCM (200 ml) at 0°C was added acetic acid (29.8 ml, 520 mmol), followed by portions of N-iodosuccinimide (16.70 g, 74.2 mmol). The resulting reaction mixture was stirred at 0° C. for 1 hour. Aqueous 10% sodium sulfite solution (200 ml) was added and stirred vigorously at room temperature for 30 minutes. The DCM layer was separated and washed with saturated sodium thiosulfate (300 ml), brine (200 ml) and then water (100 ml), and the organic layer was dried with anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give the title compound. LCMS (method E): retention time=2.37, [M+H] ⁺ =328.

Intermediate 100

5-chloro-3-iodo-2-methyl-N-(3-(1-methyl-1H-imidazol-2-yl)benzyl)pyrazolo[1,5-а]pyrimidin-7-amine

(3-(1-methyl-1H-imidazol-2-yl)phenyl)methylamine mixture (intermediate 3, 31 g, 122 mmol), 5,7-dichloro-3-iodo-2-methylpyrazolo[1,5- a]pyrimidine (intermediate 99, 39.9 g, 122 mmol) and DIPEA (63.8 ml, 365 mmol) in DMSO (500 ml) were stirred and heated at 50°C for 18 h. After cooling, the reaction mixture was poured onto ice-cold water (1500 ml). The precipitated solid was collected by filtration, dried under reduced pressure to give the title compound. LCMS (method C): retention time=2.19, [M+H] ⁺ =479.

Intermediate 101

tert-Butyl(5-chloro-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H-imidazol-2-yl)benzyl)carbamate

To a stirred solution of 5-chloro-3-iodo-2-methyl-N-(3-(1-methyl-1H-imidazol-2-yl)benzyl)pyrazolo[1,5-a]pyrimidin-7-amine (intermediate compound 100, 50 g, 96 mmol) in DCM (1000 ml) were added DIPEA (25.03 ml, 143 mmol), DMAP (1.167 g, 9.55 mmol) and di-tert-butyl dicarbonate (33.3 ml, 143 mmol). The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with dichloromethane (1000 ml), washed with water (1000 ml), dried using anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (80% ethyl acetate in petroleum ether) to give the title compound. LCMS (method D): retention time = 6.33, [M+H] ⁺ = 579.

Intermediate 102

tert-Butyl (5-chloro-3-(3-((2-hydroxyethyl)sulfonyl)-4-methoxyphenyl)-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl -1H-imidazol-2-yl)benzyl)carbamate

tert-Butyl(5-chloro-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H-imidazol-2-yl)benzyl)carbamate (intermediate) 101, 506 mg, 0.874 mmol), 2-((2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)sulfonyl)ethane-1- ol (intermediate 19, 268 mg, 0.783 mmol), triphenylarsine (13 mg, 0.042 mmol), PdCl ₂ (PhCN) ₂ (18 mg, 0.047 mmol) and sodium carbonate (193 mg, 1.821 mmol) were combined in 1.4 -dioxane (6 ml) and water (1.5 ml). The mixture was sparged with nitrogen for 1 min and heated to 80° C. for 4 h. phenyl)sulfonyl)ethan-1-ol (intermediate 19, 113 mg, 0.330 mmol), triphenylarsine (13 mg, 0.042 mmol) and PdCl ₂ (PhCN) ₂ (18 mg, 0.047 mmol). The mixture was sparged with nitrogen for 1 min and then heated to 80° C. overnight. The mixture was allowed to cool and concentrated under reduced pressure. The residue was suspended in ethyl acetate (20 ml) and filtered through Celite, washing with ethyl acetate (40 ml) followed by DCM (40 ml). The combined filtrate was concentrated under reduced pressure to remove DCM and then partitioned with water (40 ml). The organic phase was separated, passed through a hydrophobic sintered filter and concentrated under reduced pressure. The residue was dissolved in DMSO:methanol (6 ml, 1:1) and purified by MDAP chromatography (method A). Fractions containing product were combined and concentrated under reduced pressure to give the title compound. LCMS (method F): retention time = 1.12, [M+H] ⁺ = 667.

Intermediate 103

Ethyl 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)butanoate

Prepared analogously to Intermediate 79 using iodoethane to give the title compound. LCMS (method F): retention time = 1.24, [M+H] ⁺ = 559.

Intermediate 104

2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)buten- 1-ol

Prepared analogously to intermediate 81 using ethyl 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidin-5-yl)butanoate (intermediate 103) to give the title compound. LCMS (method F): retention time = 1.05, [M+H] ⁺ = 517.

Intermediate 105

3-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)pentan- 3-ol

Ethyl 3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidine-5-carboxylate (Intermediate 42 , 253 mg, 0.539 mmol) was stirred in THF (5 ml) at 0°С under nitrogen atmosphere. Ethylmagnesium bromide (1 M in THF) (2.4 mL, 2.4 mmol) was added over 5 min and the mixture was stirred at 0°C for 30 min, then warmed to room temp. After 2 h, the reaction mixture was quenched with saturated aqueous ammonium chloride (2 ml) and separated with dichloromethane (10 ml) and saturated aqueous ammonium chloride (10 ml). The organic phase was collected and the aqueous washed with dichloromethane (2×5 ml). The combined organic layers were passed through a hydrophobic sintered filter and evaporated to dryness. The residue was stirred in THF (5 ml) under nitrogen at 0°C. Ethylmagnesium bromide (1 M in THF) (1.6 ml, 1.6 mmol) was added over 5 min and the mixture was stirred at 0°C for 30 min. The mixture was quenched with saturated aqueous ammonium chloride (2 ml) and separated with dichloromethane (10 ml) and saturated aqueous ammonium chloride (10 ml). The organic phase was collected and the aqueous washed with dichloromethane (2×5 ml). The combined organic layers were passed through a hydrophobic sintered filter and concentrated under reduced pressure. The residue was dissolved in methanol:DMSO (2 ml, 1:1) and purified by MDAP chromatography (method A). Product-containing fractions were combined, evaporated to dryness and then dried in a vacuum oven overnight to give the title compound. LCMS (method J): retention time = 0.80, [M+H] ⁺ = 483.

Intermediate 106

3-bromo-N-methoxy-N,2-dimethyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-а]pyrimidine-5-carboxamide

A suspension of N,O-dimethylhydroxylamine hydrochloride (1.62 g, 16.61 mmol) in THF (10 ml) was cooled to -10°C). n-Butyl lithium (2.5 M in hexanes) (13 mL, 32.5 mmol) was added dropwise over 40 min keeping the reaction temperature below 5°C. The mixture was then stirred in an ice bath for 15 minutes. Ethyl 3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5- carboxylate (intermediate 42, 1.5 g, 3.2 mmol) in THF (10 mL). During the addition, the temperature of the mixture was maintained below 0°C. After the addition was complete, the mixture was stirred in an ice bath (at -8° C.) for 20 minutes and then allowed to warm to room temperature and stirred for 1 hour. Saturated water was added. ammonium chloride solution (20 ml) and the mixture was stirred for 30 minutes. DCM (50 ml) was added. The solution was sonicated and transferred to a separating funnel. The remaining solid was suspended in a mixture of water (50 ml) and DCM (50 ml) and sonicated. The mixture was added to a separating funnel. The organic phase was separated, and the aqueous was further extracted with dichloromethane (100 ml × 2). The combined organics were passed through a hydrophobic sintered filter and concentrated under reduced pressure. The residue was dissolved in DCM and purified by silica gel chromatography eluting with 30-100% ethyl acetate:ethanol (3:1, v/v containing 1% triethylamine) in cyclohexane (10 column volumes). Product-containing fractions were combined and concentrated under reduced pressure to give the title compound. LCMS (method J): retention time = 0.63, [M+H] ⁺ = 484.

Intermediate 107

1-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)propan- 1-he

3-bromo-N-methoxy-N,2-dimethyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-а]pyrimidine-5-carboxamide (intermediate compound 106, 530 mg, 1.094 mmol) was dissolved in THF (20 ml) and cooled to 0°C. Ethyl magnesium bromide (1 M in THF) (2.7 mL, 2.7 mmol) was added slowly. After 20 minutes, the mixture was quenched with saturated aqueous ammonium chloride (5 ml) and stirred vigorously for 10 minutes. The suspension was partitioned between water (50 ml) and DCM (50 ml). The separated aqueous phase was washed with additional DCM (2×50 ml) and the combined organic phases were passed through a hydrophobic sintered filter and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (C ₁₈ , 60 g) eluting with 30-85% acetonitrile in 10 mM ammonium bicarbonate in water (adjusted to pH 10 with ammonia) (25 column volumes). Fractions containing the desired product were combined and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (C ₁₈ , 40 g) eluting with 20-50% acetonitrile in water (containing 0.1% formic acid). Fractions containing the target product were combined and neutralized using saturated water. sodium bicarbonate, then was extracted with dichloromethane (2×50 ml). The combined organics were passed through a hydrophobic sintered filter and dried under reduced pressure to give the title compound. LCMS (method J): retention time = 0.84, [M+H] ⁺ = 453.

Intermediates 108 and 109

1-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)propan- 1-ol, isomer 1 and isomer 2.

1-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)propan- 1-one (intermediate 107, 164 mg, 0.362 mmol) was stirred in THF (3 ml) at 0°C under nitrogen atmosphere. DIBAL-H (1 M in THF) (0.75 ml, 0.75 mmol) was added dropwise and the mixture was stirred for 90 min. The mixture was quenched with aqueous sodium potassium tartrate (1 M) and stirred vigorously for 1 hour. The mixture was separated with dichloromethane (20 ml) and water (20 ml). The separated aqueous phase was washed with additional dichloromethane (2×20 ml) and the combined organics were passed through a hydrophobic sintered filter and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (C ₁₈ , 30 g) eluting with 10-40% acetonitrile (containing 0.1% formic acid) in water (containing 0.1% formic acid) (10 column volumes). Fractions containing the desired product were combined and made basic with saturated aqueous sodium bicarbonate, then extracted with DCM (2×50 ml). The combined organic layers were passed through a hydrophobic sintered filter and concentrated under reduced pressure. The residue was purified using a Chiralpak AD-H column (30 mm × 250 mm, 5 µm) eluting with 30% ethanol (containing 0.2% isopropylamine) in hexane (containing 0.2% isopropylamine) to give the title compounds. Isomer 1: LCMS (method J): retention time = 0.61, [M+H] ⁺ = 455. Chiral HPLC, retention time 5.95, 100%. Isomer 2: LCMS (method J): retention time = 0.61, [M+H] ⁺ = 455. Chiral HPLC, retention time 9.02, 100%.

Intermediate 110

1-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)ethan- 1-he

Prepared analogously to Intermediate 107 using methyl magnesium bromide to give the title compound. LCMS (method J): retention time = 0.73, [M+H] ⁺ = 439.

Intermediates 111 and 112

1-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)ethan- 1-ol, isomer 1 and isomer 2

Prepared analogously to intermediate 108 and 109 to give the title compounds. Isomer 1: LCMS (method F): retention time = 0.95, [M+H] ⁺ = 441. Chiral HPLC, retention time 7.37, 100%. Isomer 2: LCMS (method F): retention time = 0.95, [M+H] ⁺ = 441. Chiral HPLC, retention time 11.07, 99.4%.

Intermediate 113

tert-Butyl(5-acetyl-3-bromo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H-imidazol-2-yl)benzyl)carbamate

1-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)ethan- 1-one (intermediate 110, 358 mg, 0.815 mmol) was suspended in THF (10 ml). DIPEA (0.21 ml, 1.202 mmol) and DMAP (10 mg, 0.082 mmol) were added, followed by Boc anhydride (0.28 ml, 1.206 mmol). The mixture was stirred and heated to 50° C. for 1.5 h. The reaction mixture was cooled to ambient temperature, diluted with water (25 ml) and ethyl acetate (25 ml). The organic phase was collected and the aqueous was washed with additional ethyl acetate (2×25 ml). The combined organics were passed through a hydrophobic sintered filter and evaporated to dryness. The residue was dissolved in a minimum amount of DCM (1.5 ml) and wet loaded onto a 40 g silica gel column. The product was eluted using 0-50% ethanol:ethyl acetate (3:1, v/v containing 1% triethylamine) in cyclohexane. Fractions containing product were combined and evaporated to dryness, then dried under high vacuum for 2 days to give the title compound. LCMS (method F): retention time = 1.35, [M+H] ⁺ = 539.

Intermediate 114

tert-Butyl (3-bromo-5-(1-((tert-butyldimethylsilyl)oxy)vinyl)-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-N-methyl-1H-imidazole -2-yl)benzyl)carbamate

tert-Butyl(5-acetyl-3-bromo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H-imidazol-2-yl)benzyl)carbamate (intermediate) 113, 456 mg, 0.676 mmol) was stirred in DCM (5 ml) and triethylamine (0.32 ml, 2.296 mmol) at 0°С under nitrogen atmosphere. TBS-OTf (0.21 ml, 0.914 mmol) was added and the mixture was allowed to warm to room temperature, at which it was stirred for 4 h. Triethylamine (0.3 ml, 2.152 mmol) was added, followed by TBS-OTf (0.25 ml, 1.089 mmol ) and left the mixture overnight. The mixture was partitioned between DCM (20 ml) and water (20 ml). The separated aqueous phase was washed with dichloromethane (2×10 ml) and the combined organic phases were passed through a hydrophobic sintered filter and concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with 0-50% ethyl acetate in cyclohexane. The residue was triturated with dichloromethane and dried under high vacuum for 2 days to give the title compound. LCMS (method F): retention time = 1.70, [M+H] ⁺ = 653.

Intermediate 115

tert-Butyl (3-bromo-5-(1-((tert-butyldimethylsilyl)oxy)cyclopropyl)-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H- imidazol-2-yl)benzyl)carbamate

Diethylzinc (1 M in heptane) (0.795 ml, 0.795 mmol) was stirred in DCM (0.5 ml) at 0°C under nitrogen atmosphere. TFA (0.061 mL, 0.795 mmol) in DCM (0.5 mL) was added very slowly dropwise, and the mixture was stirred for 10 min. Diiodomethane (0.064 ml, 0.795 mmol) in DCM (0.5 ml) was added dropwise. After 5 minutes of stirring, tert-butyl(3-bromo-5-(1-((tert-butyldimethylsilyl)oxy)vinyl)-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1 -methyl-1H-imidazol-2-yl)benzyl)carbamate (intermediate 114, 260 mg, 0.398 mmol) in DCM (2 ml). After 1 h, the mixture was allowed to warm up to ambient temperature. After 6 hours, saturated aq. was added. ammonium chloride solution (1 ml) and the mixture was stirred vigorously for 30 minutes. Saturation was added. water. sodium bicarbonate (2 ml) and stirred the mixture for another 30 minutes. The mixture was partitioned between DCM (20 ml) and saturated aqueous ammonium chloride (20 ml). The separated aqueous phase was washed with dichloromethane (2×10 ml) and the combined organic phases were passed through a hydrophobic sintered filter and concentrated under reduced pressure. The crude material was combined with crude material from a similar reaction using tert-butyl(3-bromo-5-(1-((tert-butyldimethylsilyl)oxy)vinyl)-2-methylpyrazolo[1,5-a]pyrimidin-7-yl )(3-(1-methyl-1H-imidazol-2-yl)benzyl)carbamate (intermediate 114, 51 mg, 0.078 mmol) and purified by MDAP chromatography (method A). Product-containing fractions were combined, concentrated under reduced pressure and dried under high vacuum to give the title compound. LCMS (method F): retention time = 1.72, [M+H] ⁺ = 667.

Intermediate 116

Methyl 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate

Methyl 5-bromo-2-methylbenzoate (50 g, 218 mmol), bis (pinacolato) diboron (55.4 g, 218 mmol), potassium acetate (64.3 g, 655 mmol) and bis (triphenylphosphine) palladium (II) chloride (12.26 g, 17.46 mmol) in 1,4-dioxane (500 ml) was heated to 100° C. under nitrogen for 2 h. After cooling, the mixture was filtered through celite and washed with ethyl acetate (500 ml). A 1 M aqueous hydrochloric acid solution (500 ml) was added to the filtrate, and the phases were separated. The organic phase was washed with 1 M aqueous hydrochloric acid (250 ml), brine (250 ml), dried with magnesium sulfate and the solvent was removed under vacuum. The crude product was purified by silica gel column chromatography eluting with 0-50% ethyl acetate in cyclohexane to give the title compound. LCMS (method J): retention time = 1.34, [M+H] ⁺ = 277.

Intermediate 117

Methyl 5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidin-3-yl)-2-methylbenzoate

2-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)propan- 2-ol (intermediate 43, 25.77 g, 56.6 mmol), methyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (intermediate 116, 23.44 g, 85 mmol), potassium phosphate (18.02 g, 85 mmol), XPhos (2.70 g, 5.66 mmol) and XPhos Pd G2 (4.45 g, 5.66 mmol) were combined in 1,4-dioxane (300 ml) and water (100 ml). The mixture was treated with three vacuum/nitrogen cycles and then stirred and heated to 100° C. for 2 hours. After cooling, the mixture was partitioned between ethyl acetate (250 ml) and 2M aqueous hydrochloric acid (250 ml). The separated organic phase was washed with 2M aqueous hydrochloric acid (250 ml). The basicity of the combined aqueous phases was raised with 1M aqueous sodium hydroxide to ~pH 10 and extracted with ethyl acetate (2×250 ml). The organics were washed with brine (100 ml), dried with magnesium sulfate, and concentrated under reduced pressure to give the title compound. LCMS (method J): retention time = 0.78, [M+H] ⁺ = 525.

Intermediate 118

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-2-methylbenzoic acid

Methyl 5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidin-3-yl)-2-methylbenzoate (intermediate 117, 32.66 g, 62.3 mmol) was stirred in methanol (300 ml). 10 M aqueous sodium hydroxide solution (31.1 ml, 311 mmol) was added and the reaction mixture was stirred at 50° C. for 6 h. After cooling, ethyl acetate (500 ml) and water (500 ml) were added and the phases were separated. The organic phase was washed with 1 M aqueous sodium hydroxide solution (2×100 ml). The combined aqueous phases were neutralized to ~pH 5-6 using 2 M aqueous hydrochloric acid and extracted with ethyl acetate (4×500 ml). The combined organic layers were concentrated under reduced pressure to give the title compound. LCMS (method J): retention time = 0.65, [M+H] ⁺ = 511.

Intermediate 119

Methyl-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5- a]pyrimidin-3-yl)-2-methoxybenzoate

2-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)propan- 2-ol (intermediate 43.10 g, 21.96 mmol), methyl 2-methoxy-5-(4,4,5,5-tetramethyl1,3,2-dioxaborolan-2-yl)benzoate (intermediate 78, 9.62 g, 32.9 mmol), potassium phosphate (6.99 g, 32.9 mmol), XPhos (1.047 g, 2.196 mmol) and XPhos Pd G2 (1.728 g, 2.196 mmol) in 1,4-dioxane (100 ml) and water (33.3 ml ) was degassed (vacuum/nitrogen×3). The reaction mixture was stirred under nitrogen at 100° C. for 2 hours. The reaction mixture was allowed to cool and partitioned between ethyl acetate (250 ml) and 2M hydrochloric acid (200 ml). The organic phase was washed with 2 M hydrochloric acid (100 ml). The basicity of the combined aqueous phases was raised with 1M aqueous sodium hydroxide to pH ~10 and extracted with ethyl acetate (250 ml). The organic phase was washed with brine and dried with magnesium sulfate. The solvent was evaporated in vacuo to give the title compound. LCMS (method J): retention time = 0.63, [M+H] ⁺ = 541.

Intermediate 120

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-2-methoxybenzoic acid

To a solution of methyl 5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1, 5-a]pyrimidin-3-yl)-2-methoxybenzoate (intermediate 119, 13.55 g, 25.06 mmol) in methanol (100 ml) was added 1 M aqueous sodium hydroxide solution (12.53 ml, 125 mmol) and the reaction mixture was stirred at 50° C. for 6 hours. The reaction mixture was allowed to cool, then diluted with ethyl acetate (200 ml) and water (200 ml) and then filtered. Separate the phases. The aqueous phase was neutralized to pH ~5-6 using 2 M hydrochloric acid and then extracted using ethyl acetate (3 times). Solid sodium chloride was added to the aqueous phase. This mixture was extracted with ethyl acetate (2×150 ml). The combined ethyl acetate extracts were dried over magnesium sulfate. The solvent was evaporated in vacuo to give the title compound. LCMS (method J): retention time = 0.57, [M+H] ⁺ = 527.

Intermediate 121

Ethyl 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)- 2-(Tetrahydro-2H-pyran-4-yl)acetate

Ethyl 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidine-5 -yl)acetate (intermediate 77, 255 mg, 0.481 mmol) in THF (2 mL) under nitrogen at 0°C. 4-iodotetrahydro-2H-pyran (0.06 ml, 0.503 mmol) was added and the mixture was stirred at 0°C for 1 min, after which LiHMDS (1Mb THF) (1 ml, 1 mmol) was added. The reaction mixture was stirred at 0°C for 10 min, then allowed to warm to room temperature, and then heated to 40°C for 2 days. The reaction mixture was diluted with saturated aqueous ammonium chloride (3 ml) and stirred for 5 minutes. The suspension was transferred to a separating funnel with dichloromethane (20 ml) and water (20 ml). The phases were separated and the organic layer was collected. The aqueous layer was then washed with additional dichloromethane (20 ml) and the combined organic layers were washed with brine (20 ml), then filtered through a hydrophobic sintered filter and evaporated to dryness. The residue was loaded from DCM (1 ml) onto a silica gel (40 g) column and eluted with 10-60% ethanol:ethyl acetate (3:1, v/v containing 1% triethylamine) in cyclohexane. Product-containing fractions were combined and evaporated to dryness to give the title compound. LCMS (method F): retention time = 1.14, [M+H] ⁺ = 615.

Intermediate 122

3-iodo-2-methyl-N-(3-(1-methyl-1H-imidazol-2-yl)benzyl)-5-((tetrahydro-2H-pyran-4-yl)methyl)pyrazolo[1,5 -a]pyrimidine-7-amine

Ethyl 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)-2 -(tetrahydro-2H-pyran-4-yl)acetate (intermediate 121, 158 mg, 0.188 mmol) was dissolved in THF (2 ml) and 2 M sodium hydroxide (aq.) (1 ml, 2 mmol) and heated to 110° C. in a sealed tube for 6 hours. After cooling, DCM (20 ml) and water (20 ml) were added and the aqueous phase was neutralized to pH 7 with 2 M aq. HCl solution. The phases were separated and the organic layer was collected. The aqueous layer was then washed with additional dichloromethane (2 x 10 ml) and the combined organic layers were passed through a hydrophobic sintered filter and evaporated to dryness to give the title compound. LCMS (method F): retention time = 1.07, [M+H] ⁺ = 543.

Intermediate 123

N-(3-hydroxypropyl)-N-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoic acid (245 mg, 0.988 mmol) and HATU (413 mg, 1.086 mmol) and DIPEA (0.35 ml, 2.004 mmol) was stirred in THF (5 ml) in an air atmosphere at ambient temperature. After 10 minutes, 3-(methylamino)propan-1-ol (0.12 ml, 1.234 mmol) was added and the mixture was stirred overnight. The mixture was partitioned between saturated aqueous sodium bicarbonate (30 ml) and ethyl acetate (20 ml). The separated aqueous phase was washed with additional ethyl acetate (2×20 ml) and the combined organics washed with brine (20 ml). The organic layer was passed through a hydrophobic sintered filter and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with 40-100% ethyl acetate:ethanol (3:1 containing 1% triethylamine) in cyclohexane to give the title compound. LCMS (method J): retention time = 0.93, [M+H] ⁺ = 320.

Connection examples

Compound 1

1-(3-(4-methoxy-3-(methylsulfonyl)phenyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5 -a]pyrimidin-5-yl)ethanol

3-(4-methoxy-3-(methylsulfonyl)phenyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidine-5-carbaldehyde (intermediate 12, 30 mg, 0.057 mmol) was dissolved in THF (2.5 ml) and cooled to 0°C. Methylmagnesium bromide (3 M in diethyl ether) (0.1 ml, 0.3 mmol) was added dropwise and the reaction mixture was allowed to warm to room temperature and stirred for another 3 h. Additional methylmagnesium bromide (3 M in diethyl ether) (0.1 ml, 0.3 mmol) was added and the reaction mixture was stirred for another 20 h. The reaction was quenched by adding 1 M HCl (10 ml) and then extracted with ethyl acetate (3×10 ml). The combined organic phases were dried through a hydrophobic sintered filter and the solvent was removed under vacuum. The residue was purified by MDAP chromatography (Method A) to give the title compound. LCMS (method F): retention time = 0.94, [M+H] ⁺ = 547.

Compound 2

2-((5-(5-(1-hydroxyethyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-2-methoxyphenyl)sulfonyl)ethan-1-ol

tert-Butyl(5-formyl-3-(3-((2-hydroxyethyl)sulfonyl)-4-methoxyphenyl)-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl -1H-imidazol-2-yl)benzyl)carbamate (intermediate 20, 100 mg, 0.151 mmol) in THF (3 ml) was stirred at 0°C under nitrogen, then 3.4 M methylmagnesium bromide in 2-methyltetrahydrofuran was added in one portion (0.1 ml). The reaction mixture was allowed to cool to room temperature, and the reaction mixture was stirred under nitrogen atmosphere for 1 hour. The reaction mixture was fumed with water (10 ml) and stirred under nitrogen atmosphere for 5 minutes. The solvent was removed under vacuum. The crude material was dissolved in DCM (20 ml) and separated with water (15 ml). The organic layer was separated and the aqueous phase was extracted with additional DCM (2×20 ml). The combined organic layers were passed through a hydrophobic sintered filter and the solvent was removed under vacuum. The crude material was dissolved in methanol (3 ml) and 4 M HCl in 1,4-dioxane (2 ml, 8 mmol) was added. The reaction mixture was stirred under nitrogen for 5 hours. The solvent was removed in vacuo. The crude material was dissolved in methanol:DMSO (2×1 ml, 1:1, v/v) and purified by MDAP chromatography (method A) to give the title compound. LCMS (method F): retention time = 0.86, [M+H] ⁺ = 577.

The following compounds were prepared in a manner analogous to the preparation of compound 2 using the following Grignard reagents:

1 M Ethylmagnesium bromide in THF,

0.5 M cyclopropylmagnesium bromide in THF,

1 M isopropylmagnesium bromide in THF.

Compound 6

2-(3-(3-((2-hydroxyethyl)sulfonyl)-4-methoxyphenyl)-2-methyl-7-((3-(1-1etyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo [1,5-a]pyrimidin-5-yl)propan-2-ol

2-((2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)sulfonyl)ethan-1-ol (intermediate 19, 121 mg, 0.354 mmol), sodium carbonate (87 mg, 0.821 mmol), PdCl ₂ (dppf) (19 mg, 0.026 mmol), water (1 ml) and 2-(3-iodo-2-methyl-7-((3- (1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)propan-2-ol (intermediate 21, 136 mg, 0.271 mmol) in isopropyl alcohol (1 ml) were combined and heated at 120° C. in a microwave reactor for 1.5 h. The reaction mixture was filtered through celite (washing with dichloromethane) and evaporated to dryness. The filtrate was then partitioned between water (20 ml) and DCM (20 ml). The organic phase was collected and the aqueous washed with dichloromethane (2×10 ml). The combined organic layers were passed through a hydrophobic sintered filter and evaporated to dryness. The residue was dissolved in DMSO:methanol (2 ml) and purified by MDAP chromatography (method A) to give the title compound. LCMS (method F): retention time = 0.91, [M+H] ⁺ = 591.

Compound 7

2-chloro-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1, 5-a]pyrimidin-3-yl)-N-(3-hydroxypropyl)-N-methylbenzamide

2-chloro-N-(3-hydroxypropyl)-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 23, 105 mg, 0.297 mmol), sodium carbonate (69.6 mg, 0.657 mmol), PdCl ₂ (dppf)-DCM (12.5 mg, 0.015 mmol), water (1 ml) and 2-(3-iodo-2-methyl-7-(( 3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)propan-2-ol (intermediate 21, 110 mg, 0.219 mmol) in isopropyl alcohol (1 ml) was combined and heated at 120° C. in a microwave reactor for 2 hours. More 2-chloro-N-(3-hydroxypropyl)-N-methyl-5-(4,4,5,5- tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 23, 105 mg, 0.297 mmol) and the reaction mixture was heated at 120°C in a microwave reactor for 2 h. The reaction mixture was filtered through celite using DCM ( 20 ml). The filtrate was washed with water (20 ml) and the aqueous phase was extracted with additional DCM (2×20 ml). The combined organic layers were passed through a hydrophobic sintered filter and the solvent was removed under reduced pressure. The residue was dissolved in methanol:DMSO (0.6 ml, 1:1, v/v) and purified by MDAP chromatography (method A). The residue was loaded into DCM (2 ml) and passed through a silica gel cartridge (1 g) eluting with ethyl acetate:ethanol (3:1, v/v containing 1% triethylamine). The residue was taken up in DCM (2 ml) and purified by chromatography on silica gel (4 g) eluting with ethyl acetate:ethanol (3:1, v/v containing 1% triethylamine) in cyclohexane (0%, 2 column volumes). ; 0-100%, 5 column volumes; 100%, 7 column volumes) to give the title compound. LCMS (method F): retention time = 0.98, [M+H] ⁺ = 602.

Compound 8

(S)-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1, 5-a]pyrimidin-3-yl)-2-methoxyphenyl)3-hydroxypyrrolidin-1-yl)methanone

Prepared in a manner analogous to Compound 7 using (S)-(3-hydroxypyrrolidin-1-yl)(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)phenyl)methanone (intermediate 25) to give the title compound. LCMS (method F): retention time = 0.88, [M+H] ⁺ = 596.

Compound 9

5-(5-(1-hydroxyethyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-3- yl)-N-(3-hydroxypropyl)-2-methoxy-N-methylbenzamide

N-(3-hydroxypropyl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 28, 80 mg, 0.160 mmol) in isopropyl alcohol (0.5 ml), tert-butyl (5- (1-hydroxyethyl) -3-iodo-2-methylpyrazolo[1,5-a] pyrimidin-7-yl) (3- (1-methyl -1H-imidazol-2-yl)benzyl)carbamate (intermediate 26, 80 mg, 0.136 mmol), PdCl ₂ (dppf) (9 mg, 0.012 mmol), sodium carbonate (45 mg, 0.425 mmol) and water (0.5 ml) were combined and heated at 120°C for 1.5 h in a microwave reactor. The reaction mixture was diluted in DCM (10 ml) and water (10 ml) and the phases were separated. The organic layer was collected and the aqueous was further washed with dichloromethane (2×10 ml) and the combined organics were passed through a hydrophobic sintered filter and evaporated to dryness. The residue was dissolved in DMSO:methanol (1:1, v/v, 1 ml) and purified by MDAP chromatography (method A). Fractions containing the product were combined and evaporated to dryness. The residue was purified by silica gel column chromatography eluting with 50-100% ethyl acetate:ethanol (3:1, v/v containing 1% triethylamine) in cyclohexane to give the title compound. LCMS (method F): retention time = 0.87, [M+H] ⁺ = 584.

The following compounds were prepared in a manner analogous to compound 9 using the following boronic acid esters:

2-chloro-N-(3-hydroxypropyl)-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 23), N- ethyl-N-(2-hydroxyethyl)-2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 30),

Compound 12

5-(5-(1-hydroxyethyl)-2-methyl-7-((3-(1-methyl-1H-imidaeol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-3- yl)-2-methoxy-N-methyl-N-(tetrahydro-2H-pyran-4-yl)benzamide

tert-Butyl (5-(1-hydroxyethyl)-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H-imidazol-2-yl)benzyl) carbamate (intermediate 26, 70 mg, 0.119 mmol), 2-methoxy-N-methyl-N-(tetrahydro-2H-pyran-4-yl)-5-(4,4,5,5-tetramethyl-1, 3,2-dioxaborolan-2-yl)benzamide (intermediate 34, 77 mg, 0.143 mmol), PdCl ₂ (dppf) (9 mg, 0.012 mmol), sodium carbonate (41 mg, 0.387 mmol), isopropyl alcohol (2 ml) and water (1 ml) were combined and heated in a microwave reactor at 120°C for 1.5 h. The reaction mixture was filtered using celite, washed with methanol (100 ml) and the solvent was removed under vacuum. The residue was dissolved in DCM (20 ml) and separated with water (10 ml). The organic layer was separated and the aqueous phase was extracted with additional DCM (2×10 ml). The combined organic layers were passed through a hydrophobic sintered filter and the solvent was removed under vacuum. The residue was dissolved in methanol (2 ml) and 4 M HCl in 1,4-dioxane (2 ml) was added. The reaction mixture was stirred under nitrogen at room temperature for 5 hours. The solvent was removed in vacuo. The residue was dissolved in water (15 ml), neutralized with 2 M sodium hydroxide and separated with dichloromethane (20 ml). The organic layer was separated and the aqueous phase was extracted with additional DCM (2×20 ml). The combined organic layers were passed through a hydrophobic sintered filter and the solvent was removed under vacuum. The sample was dissolved in methanol:DMSO (1 ml, 1:1, v/v) and purified by MDAP chromatography (method B). The solvent was removed under vacuum. The residue was triturated with ethyl acetate then diethyl ether to give the title compound. LCMS (method F): retention time = 0.94, [M+H] ⁺ = 610.

The following compounds were prepared in a manner analogous to compound 12 by varying the organic reaction solvent between 1,4-dioxane isopropyl ester and using boronic acid esters:

N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 36),

(S)-(2-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)(3-hydroxypyrrolidin-1-yl)methanone (intermediate 38 ),

2-chloro-N-ethyl-N-(2-hydroxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 32),

(S)-N-(1-hydroxypropan-2-yl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 40),

Compound 17

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-а] pyrimidin-3-yl)-N-(3-hydroxypropyl)-N,2-dimethylbenzamide

N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 36, 542 mg, 0.813 mmol ), 2-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl) propan-2-ol (intermediate 43, 333 mg, 0.622 mmol), PdCl ₂ (dppf) (47 mg, 0.064 mmol), sodium carbonate (198 mg, 1.865 mmol), 1,4-dioxane (3 ml) and water (1 ml) was combined and heated at 120°C for 1.5 h in a microwave reactor. N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 36, 262 mg, 0.393 mmol) and PdCl ₂ (dppf) (20 mg, 0.027 mmol), and the reaction mixture was heated at 120°С for 1.5 h in a microwave reactor. The reaction mixture was diluted in DCM (20 ml) and water (20 ml) and the phases were separated. The organic layer was collected and the aqueous layer was further washed with dichloromethane (2×20 ml) and the combined organics were passed through a hydrophobic sintered filter, then evaporated to dryness. The residue was dissolved in DMSO:methanol (4 ml) and purified by MDAP chromatography (method A). Fractions containing the product were combined and evaporated to dryness. The residue was purified by silica gel column chromatography eluting with 20-100% ethyl acetate:ethanol (3:1, v/v containing 1% triethylamine) in cyclohexane. Product-containing fractions were combined, evaporated to dryness, and then dried on a high vacuum line for 3 days to give the title compound. LCMS (method F): retention time = 0.96, [M+H] ⁺ = 582.

The following compounds were prepared in a manner analogous to compound 17 using either 1,4-dioxane or isopropyl alcohol as reaction solvent and the following boronic acid esters:

(S)-(2-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)(3-hydroxypyrrolidin-1-yl)methanone (intermediate 38 ),

N-ethyl-N-(2-hydroxyethyl)-2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 30),

(S)-N-(1-hydroxypropan-2-yl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 40),

N-(tetrahydro-2H-pyran-4-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 34),

(S)-N-(1-hydroxypropan-2-yl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate connection 45),

2-methoxy-N,N-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 47),

(R)-(2-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)(2-(hydroxymethyl)pyrrolidin-1-yl)methanone ( intermediate compound 49),

(R)-(2-(hydroxymethyl)pyrrolidin-1-yl)(2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone ( intermediate connection 51),

N-(2-hydroxyethyl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 53),

(3-hydroxy-3-methylpyrrolidin-1-yl)(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone (intermediate 55 ),

(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)(1,4-oxazepan-4-yl)methanone (intermediate 57),

(S)-(3-hydroxypyrrolidin-1-yl)(2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone (intermediate 59 ),

(S)-(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)(3-methylmorpholino)methanone (intermediate 61),

(3-Hydroxy-3-methylpyrrolidin-1-yl)(2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone (Intermediate 63 ),

N-(3-hydroxypropyl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 28)

The following compounds were prepared in a manner analogous to compound 17 using intermediate 65 and either 1,4-dioxane or isopropyl alcohol as reaction solvent and a temperature between 100°C and 120°C and the following boronic acid esters:

N-(3-hydroxypropyl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 28),

N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 36),

(R)-(2-(hydroxymethyl)pyrrolidin-1-yl)(2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone ( intermediate connection 51).

The following compounds were prepared in a manner analogous to compound 17 using the following boronic acid esters:

(R)-(2-(hydroxymethyl)pyrrolidin-1-yl)(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone ( intermediate compound 67).

Alternative method for obtaining compound 17 - 5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[ 1,5-a]pyrimidin-3-yl)-N-(3-hydroxypropyl)-N,2-dimethylbenzamide

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-2-methylbenzoic acid (intermediate 118, 12 g, 23.5 mmol), 3-(methylamino)propan-1-ol (2.81 ml, 29.4 mmol) and DIPEA (8.21 ml, 47.0 mmol) were stirred in THF (100 ml). HATU (11.17 g, 29.4 mmol) and DMF (10 ml) were added and the mixture was stirred under nitrogen at room temperature for 3 h. The mixture was partitioned between ethyl acetate (400 ml) and saturated aqueous sodium bicarbonate (400 ml). The aqueous phase was back extracted with ethyl acetate (200 ml). The combined organic phases were washed with brine (200 ml), dried with magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography eluting with 0-25% ethanol in ethyl acetate (5 column volumes) and 25% ethanol in ethyl acetate (7 column volumes). Fractions containing the product were combined, evaporated in vacuo and purified by the same chromatographic method until a material content of >97.5% (according to HPLC) was reached. The residue was triturated with diethyl ether and the solvent was evaporated under vacuum. Ethyl acetate (40 ml) was added and the mixture was stirred at room temperature for 5 days.

The resulting white solid was collected by filtration, washed with ethyl acetate (25 ml) and dried under vacuum at 40° C. to give the title compound. LCMS (method M): retention time = 2.49, [M+H] ⁺ = 582. δ _n (400 MHz, d ₆ -DMSO) (Rotameter mix) 8.53 (t, J=6.5 Hz, 1H), 7.73 (s , 1Н), 7.71-7.67 (m, 0.75Н), 7.65 (s, 0.75Н), 7.61 (d, J=1.3 Hz, 0.5Н), 7.59-7.54 (m, 1Н), 7.48-7.42 (m, 2Н), 7.33-7.27 (m, 1Н), 7.22 (d, J=1.0 Hz, 1Н), 6.94 (d, J=1.0 Hz, 1Н), 6.45 (s, 1Н), 5.18 (s, 1Н), 4.70 (d, J=6.6 Hz, 2H), 4.47 (t, J=5.3 Hz, 0.5H), 4.34 (t, J=4.9 Hz, 0.5H), 3.70 (s, 3H), 3.56-3.46 (m , 2Н), 3.28-3.20 (m, 2Н), 3.00 (s, 1.5Н), 2.82 (s, 1.5Н), 2.58 (s, 3Н), 2.21 (app. d, J=5.1 Hz, 3Н), 1.79-1.70 (m, 1H), 1.66-1.57 (m, 1H), 1.39 (s, 6H) ppm. Δ _C (151 MHz, D ₆ -DMSO) (mixture of rotamers) 170.3, 170.2, 169.0, 150.3, 150.2, 146.39, 146.36, 146.3, 145.4, 138.6, 137.0, 136.9, 130.8, 130.14, 130.05, 130.0, 128.6, 127.5.5.5.5.5.5.5.5.5.5.5.5 127.4, 126.89, 126.86, 126.81, 124.84, 124.79, 123.4, 104.72, 104.67, 81.55, 81.53, 72.39, 72.38, 58.4, 58.0, 47.4, 44.4.4.4.5, 36.3, 34.3, 31.8, 31.0, 30.2.0, 30.0.0, 30.0.0.0, 30.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0EN , 18.1, 14.7, 14.6 ppm. High resolution mass spectroscopy (electrospray ionization) calc. for C ₃₃ H ₃₉ N ₇ O ₃ +H ⁺ 582.3193, exp. 582.3190 [M+H] ⁺ .

Alternative method for preparing compound 22

(S)-N-(1-hydroxypropan-2-yl)-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazole- 2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-N,2-dimethylbenzamide

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-2-methylbenzoic acid (intermediate 118, 7.55 g, 14.79 mmol), (S)-2-(methylamino)propan-1-ol (1.977 g, 22.18 mmol) and DIPEA (5.17 ml, 29.6 mmol) was stirred in THF (50 ml). DMF (2.5 ml) and HATU (7.03 g, 18.48 mmol) were added and stirred under nitrogen at room temperature for 20 h. The mixture was partitioned between ethyl acetate (200 ml) and saturated aqueous sodium bicarbonate (200 ml). The aqueous phase was extracted with ethyl acetate (100 ml). The combined organic phases were washed with brine (100 ml), dried over magnesium sulfate and the solvent removed in vacuo. The residue was chromatographed on an amine-capped silica gel column (KP-NH) eluting with 0-100% ethyl acetate in cyclohexane. The mixture was then purified by reverse phase chromatography eluting with 5-50% acetonitrile (containing ammonia) in water (10 mM ammonium bicarbonate/ammonia). The resulting residue was triturated in methanol and then diethyl ether (50 ml), in which the mixture was rapidly stirred for 30 minutes. The solvent was removed under vacuum, the residue was dissolved in ethyl acetate (70 ml) and stirred at high speed for 24 h. The solid was collected by filtration, washed with ethyl acetate, dried under vacuum at 40°C. LCMS (method M): retention time = 2.56, [M+H] ⁺ = 582. δ _n (400 MHz, d ₆ -DMSO) (Mixed rotamers) 8.58-8.49 (m, 1H), 7.81 (d, J= 8.1 Hz, 0.3H), 7.73 (s, 1H), 7.71-7.62 (m, 1H), 7.59-7.51 (m, 1.7H), 7.49-7.42 (m, 2H), 7.32-7.25 (m, 1H) , 7.22 (s, 1Н), 6.97-6.92 (m, 1Н), 6.49-6.42 (m, 1Н), 5.17 (s, 1Н), 4.80 (t, J=5.3 Hz, 0.7Н), 4.76 (t, J=5.5Hz, 0.3H), 4.70(d, J=6.2Hz, 2H), 3.74-3.64(m, 3.5H), 3.64-3.57(m, 0.5H), 3.53-3.39(m, 1.3H ), 3.29-3.23 (m, 0.7H), 2.90-2.84 (m, 2H), 2.68 (s, 1H), 2.60-2.56 (m, 3H), 2.25-2.18 (m, 3H), 1.41-1.35 ( m, 6Н), 1.10 (d, J=7.0 Hz, 1Н), 1.06-1.00 (m, 2Н) ppt. Δ _s (176 MHz, D ₆ -DMSO, 120 ° C) 170.2, 168.3, 149.8, 146.1, 146.0, 145.0, 137.9, 137.2, 130.7, 130.2, 129.3, 128.0, 127.1, 126.6, 126.50, 126.47, 124.5, 122.6, 122.6 , 104.8, 81.2, 71.8, 62.0, 44.5, 33.5, 29.5, 17.5, 13.8 ppm. High resolution MS (electrospray ionization), calc. for C ₃₃ H ₃₉ N ₇ O ₃ +H ⁺ 582.3193, exp. 582.3195 [M+H] ⁺ .

Alternative method for preparing compound 36

(R)-(2-(Hydroxymethyl)pyrrolidin-1-yl)(5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazole -2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-2-methoxyphenyl)methanone

To a solution/suspension of 5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1, 5-a]pyrimidin-3-yl)-2-methoxybenzoic acid (intermediate 120, 3.0 g, 5.7 mmol), (R)-pyrrolidin-2-ylmethanol (0.576 g, 5.7 mmol) and DIPEA (1.990 ml, 11.39 mmol) in THF (100 mL) was added HATU (2.71 g, 7.12 mmol). The reaction mixture was stirred under nitrogen atmosphere at room temperature for 20 hours. The reaction mixture was partitioned between ethyl acetate (200 ml) and saturated sodium bicarbonate solution (200 ml). The organic phase was washed with brine (100 ml) and dried with magnesium sulfate.

The solvent was removed under vacuum. The residue was dissolved in DCM, applied to a silica gel cartridge (120 g) and eluted with 0-25% ethanol in ethyl acetate (5 column volumes) and 25% ethanol in ethyl acetate (6 column volumes). The desired fractions were combined and evaporated under vacuum. The residue was dissolved in DMSO:methanol (1:1 v/v) and applied to a C ₁₈ cartridge (120 g) pre-conditioned to 5% acetonitrile (containing ammonia) in water (10 mM ammonium bicarbonate containing ammonia) . Elution was then carried out with a gradient of 5-40% acetonitrile (containing ammonia) in water (10 mM ammonium bicarbonate containing ammonia). The gradient was maintained using 29% acetonitrile (containing ammonia) in water (10 mM ammonium bicarbonate containing ammonia) while the product was eluted. The desired fractions were combined and evaporated under vacuum. The residue was triturated with diethyl ether and the solvent was evaporated. Ethyl acetate (25 ml) and methanol (1 ml) were added to the residue. The mixture was subjected to temperature cycles from room temperature to 45°C and back 6 times, and then left under stirring at room temperature for 18 hours. The resulting white solid was collected by filtration and washed with ethyl acetate (25 ml). The solid was dried under vacuum at 40° C. for 20 hours to give the title compound. LCMS (method J): retention time = 0.59, [M+H] ⁺ = 610. δ _n (700 MHz, DMSO-d ₆ , 120°C) 7.94 (br s, 1H), 7.75 (s, 1H), 7.75-7.72 (m, 1H), 7.65 (br s, 1H), 7.58 (brd, J=7.3 Hz, 1H), 7.50-7.47 (m, 1H), 7.47-7.42 (m, 1H), 7.16-7.14 (m, 1Н), 7.16-7.13 (m, 1Н), 6.95 (s, 1Н), 6.44 (s, 1Н), 4.74 (s, 2Н), 4.18 (br s, 1Н), 3.85 (s, 3Н) , 3.75-3.71 (m, 1H), 3.70 (s, 3H), 3.51-3.42 (m, 1H), 3.39-3.15 (m, 2H), 2.56 (s, 3H), 1.96-1.71 (m, 2H) , 2.04-1.68 (m, 2H), 1.44 (s, 6H) ppm. δ _C-ЯМР (176 МГц, ДМСО-d ₆ , 120°С) 168.11, 166.71, 152.36, 149.48, 146.05, 146.01, 144.77, 137.90, 130.71, 128.70, 127.90, 127.35, 127.05, 126.65, 126.56, 126.44, 126.43 ppm High resolution MS (electrospray ionization), calc. for C ₃₄ H ₃₉ N ₇ O ₄ + H ⁺ 610.3063, exp. 610.3141 [M+H] ⁺ .

Alternative method for the preparation of compound 21 ,5-a]pyrimidin-3-yl)-2-methoxy-N-methyl-N-(tetrahydro-2H-pyran-4-yl)benzamide

To a solution/suspension of 5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1, 5-a]pyrimidin-3-yl)-2-methoxybenzoic acid (intermediate 120, 2.35 g, 4.46 mmol) and DIPEA (1.6 ml, 9.16 mmol) in THF (50 ml) was added HATU (2.35 g, 6.18 mmol) . The mixture was stirred at room temperature for 10 min and N-methyltetrahydro-2H-pyran-4-amine (0.514 g, 4.46 mmol) was added. The mixture was stirred under nitrogen at room temperature for 16 hours. The reaction mixture was partitioned between ethyl acetate (100 ml) sat. water. sodium bicarbonate solution (100 ml). The aqueous phase was extracted a second time with ethyl acetate (100 ml). The combined organic phases were washed with brine (100 ml) and passed through a hydrophobic sintered filter. The solvent was removed under reduced pressure. The residue was purified by silica gel chromatography (340 g). This compound was dissolved in minimal DCM (+ a few drops of methanol), loaded over the column by injection, then eluted using 0-25% ethanol in ethyl acetate (6 column volumes) and then treated with 25% ethanol in ethyl acetate (6 column volumes) . Fractions containing the desired product were combined and concentrated under reduced pressure. Approximately half of the material was dissolved in DMSO:methanol (1:1 v/v) and applied to a C ₁₈ cartridge (120 g) pre-conditioned to 5% acetonitrile (containing ammonia) in water (10 mM ammonium bicarbonate containing ammonia), then eluted with a gradient of 5-20% acetonitrile (with ammonia) in water (10 mM ammonium bicarbonate with ammonia) (6 column volumes) followed by a gradient of 20-60% acetonitrile (with ammonia) in water (10 mM ammonium bicarbonate with ammonia) (6 column volumes). The gradient was then held at 47.5% acetonitrile (with ammonia) in water (10 mM ammonium bicarbonate with ammonia) until the product eluted. The rest of the material was purified in a similar manner using a gradient of 25-55% acetonitrile (with ammonia) in water (10 mM ammonium bicarbonate with ammonia), bicarbonate/NH3). The gradient was held at 45% acetonitrile (with ammonia) in water (10 mM ammonium bicarbonate with ammonia) while the product eluted. Fractions containing pure product from both purification procedures were combined, concentrated under reduced pressure, then dried in vacuo for 24 hours to give the title compound. LCMS (method J): retention time = 0.65, [M+H] ⁺ = 624. δ _n (600 MHz, d ₆ -DMSO) (Mixed rotamers) 8.54-8.49 (m, 1H), 7.77-7.72 (m, 2Н), 7.63 (d, J=1.8 Hz, 0.5Н), 7.59-7.54 (m, 1Н), 7.53 (d, J=2.2 Hz, 0.5Н), 7.47-7.43 (m, 2Н), 7.22 (s , 1Н), 7.18-7.13 (m, 1Н), 6.94 (s, 1Н), 6.43 (s, 1Н), 5.17 (s, 1Н), 4.69 (d, J=6.2 Hz, 2Н), 4.64-4.58 ( m, 0.5H), 3.98-3.91 (m, 1H), 3.90-3.85 (m, 0.5H), 3.82-3.75 (m, 3.5H), 3.70 (s, 3H), 3.57-3.50 (m, 0.5H ), 3.43 (t, J=11.2 Hz, 1H), 3.12 (t, J=11.2 Hz, 0.5H), 3.00 (t, J=11.6 Hz, 0.5H), 2.88 (s, 1.5H), 2.69 ( s, 1.5H), 2.56 (ar. d, J=15.8 Hz, 3H), 1.89-1.72 (m, 2H), 1.59-1.51 (m, 1.5H), 1.42 (d, J=11.4 Hz, 0.5H ), 1.39-1.33 (m, 6H) ppm. δ _s (151 MHz, D ₆ -DMSO) (mixture of rotamers) 168.80, 168.77, 168.1, 167.9, 152.50, 152.47, 150.0, 149.9, 146.4, 146.34, 146.26, 145.18, 145.16, 138.6, 130.8, 129.2, 128.9, 128.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6.6AID 127.5,126.89, 126.87, 126.80, 126.7, 126.6, 126.5, 125.9, 123.4, 111.6, 111.4, 104.7, 104.5, 81.4, 81.3, 72.4, 72.3, 66.5, 66.3, 66.2, 55.7, 55.4, 54.9, 54.8.8, 49. , 44.4, 34.3, 30.5, 30.3, 09.30, 07.30, 30.0, 29.2, 26.6, 14.5, 14.4 ppm. High resolution MC (electrospray ionization), calc. for C ₃₅ H ₄₁ N ₇ O ₄ +H ⁺ 624.3220, exp. 624.3294 [M+H] ⁺ .

Compound 37

2-Chloro-N-(2-hydroxyethyl)-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl) benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-N-methylbenzamide

2-(3-Bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin- 5-yl)propan-2-ol (intermediate 43, 305 mg, 0.636 mmol), XPhos (32 mg, 0.067 mmol), XPhos Pd G2 (53 mg, 0.067 mmol) and tripotassium phosphate (443 mg, 2.087 mmol). 2-Chloro-N-(2-hydroxyethyl)-N-methyp-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (Intermediate 69) 446 mg , 0.788 mmol) was dissolved in 1,4-dioxane (7.5 ml) and added to the microwave vessel, after which water (2.5 ml) was added. The vessel was sealed and heated under microwave treatment at 100°C for 1.5 h. The reaction mixture was diluted with water (25 ml) and separated with dichloromethane (25 ml). The organic phase was collected and the aqueous phase was washed with dichloromethane (4×25 ml). The organic phases were combined and the solvent was passed through a hydrophobic sintered filter and then through a second hydrophobic sintered filter with a layer of florisil (2 cm deep). The florisil layer was washed with ethyl acetate:ethanol (3:1 containing 1% triethylamine) (25 ml). The filtrate was evaporated to dryness. The residue was dissolved in a minimum of DMSO:methanol (1:1, v/v) and purified by MDAP chromatography (method A). The desired fractions were combined and the solvent was removed. The residue was purified by silica gel column chromatography eluting with a 30-100 gradient of ethyl acetate:ethanol (3:1 containing 1% triethylamine) in cyclohexane. The desired fractions were combined and the solvent was removed. The residue was triturated with minimal diethyl ether to give the title compound. LCMS (method F): retention time = 0.96, [M+H] ⁺ = 588.

The following compounds were prepared in a manner analogous to compound 37 using the following boronic esters:

N-Ethyl-N-(2-hydroxyethyl)-2-methyl-5-(4,4,5,5-tetramethyl1,3,2-dioxaborolan-2-yl)benzamide (Intermediate 73)

(R)-N-(1-hydroxypropan-2-yl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate compound 86),

N-(1-hydroxypropan-2-yl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 88),

(3-hydroxypyrrolidin1-yl)(2-methyl-5-(4,4,5,5-tetramethyl1,3,2-dioxaborolan-2-yl)phenyl)methanone (intermediate 90),

N-(1-hydroxypropan2-yl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (Intermediate 92)

The following compounds were prepared in a manner analogous to compound 17 using either 1,4-dioxane or isopropyl alcohol as reaction solvent and a temperature of 100°C to 140°C, and the following boronic esters:

N-(2-hydroxyethyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (Intermediate 71),

(2-(Hydroxymethyl)pyrrolidin-1-yl)(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone (intermediate 84) ,

N,2-Dimethyl-N-(tetrahydro-2H-pyran-4-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (Intermediate 96 ),

The following compounds were prepared in a manner analogous to that of compound 17 using intermediate 65 and either 1,4-dioxane or isopropyl alcohol as reaction solvent, and a temperature of 100°C to 140°C, and the following boronic esters:

N-(2-hydroxyethyl)-2-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 53),

N-(2-hydroxyethyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (Intermediate 71),

2-chloro-N-(2-hydroxyethyl)-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 69),

(R)-(2-(hydroxymethyl)pyrrolidin-1-yl)(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone ( intermediate compound 67),

(S)-(3-hydroxypyrrolidin-1-yl)(2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone (intermediate 59 ),

(3-hydroxy-3-methylpyrrolidin-1-yl)(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone (intermediate 55 ),

(S)-(3-hydroxypyrrolidin-1-yl)(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone (intermediate 25 ),

Compound 53

2-(3-(3-((2-hydroxyethyl)sulfonyl)-4-methoxyphenyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo [1,5-a]pyrimidin-5-yl)propan-1-ol

Prepared analogously to compound 37 using 5-yl)propan-1-ol (intermediate 81) and 2-((2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl) sulfonyl)ethanol (intermediate 19) to give the title compound. LCMS (method J): retention time=0.53, [M+H] ⁺ =591.

Compound 54

5-(5-(1-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-N-(3-hydroxypropyl)-N,2-dimethylbenzamide

Prepared in a manner analogous to Compound 17 using 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidin-5-yl)propan-1-ol (intermediate 81) and N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2 -dioxaborolan-2-yl)benzamide (intermediate 36) to give the title compound. LCMS (method F): retention time=0.92, [M+H] ⁺ =582.

Compound 55

5-(5-(1-hydroxy-2-methylpropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1, 5-a]pyrimidin-3-yl)-N-(3-hydroxypropyl)-N,2-dimethylbenzamide

Prepared in a manner analogous to Compound 17 using 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidin-5-yl)-2-methylpropan-1-ol (intermediate 82) and N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1, 3,2-dioxaborolan-2-yl)benzamide (intermediate 36) to give the title compound. LCMS (method F): retention time=0.99, [M+H] ⁺ =596.

Compound 56

2-(3-(3-((2-hydroxyethyl)sulfonyl)-4-methoxysenyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo [1,5-a]pyrimidin-5-yl)-2-methylpropan-1-ol

Prepared in a manner analogous to Compound 17 using 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidin-5-yl)-2-methylpropan-1-ol (intermediate 82) and 2-((2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan- 2-yl)phenyl)sulfonyl)ethanol (intermediate 19) to give the title compound. LCMS (method F): retention time=0.95, [M+H] ⁺ =605.

Compound 57

2-chloro-5-(5-(1-hydroxyethyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-а] pyrimidin-3-yl)-N-(3-hydroxypropyl)-N-methylbenzamide, isomer 1

Prepared in a manner analogous to Compound 37 using tert-butyl(5-(1-hydroxyethyl)-3-iodo-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H -imidazol-2-yl)benzyl)carbamate, isomer 1 (intermediate 93) and 2-chloro-N-(3-hydroxypropyl)-N-methyl-5-(4,4,5,5-tetramethyl-1, 3,2-dioxaborolan-2-yl)benzamide (intermediate 23) to give the title compound. LCMS (method F): retention time=0.95, [M+H] ⁺ =588.

Compound 58

2-chloro-5-(5-(1-hydroxyethyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-а] pyrimidin-3-yl)-N-(3-hydroxypropyl)-N-methylbenzamide, isomer 2

Prepared analogously to compound 37 using tert-butyl (5-(1-hydroxyethyl)-3-iodo-2-methylpyrazolo[1,5-а]pyrimidin-7-yl) 2-yl)benzyl)carbamate, isomer 2 (intermediate 94) and 2-chloro-N-(3-hydroxypropyl)-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2 -dioxaborolan-2-yl)benzamide (intermediate 23) to give the title compound. LCMS (method F): retention time=0.95, [M+H] ⁺ =588.

Compound 59

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-N,2-dimethyl-N-(tetrahydrofuran-3-yl)benzamide

A solution of 5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidin-3-yl)-2-methylbenzoic acid (intermediate 118, 49 mg, 0.096 mmol), HATU (40.1 mg, 0.106 mmol) and DIPEA (0.034 ml, 0.192 mmol) in DMF (0.4 ml) was stirred for 20 min at room temp. N-Methyltetrahydrofuran-3-amine (0.013 ml, 0.115 mmol) was added to the reaction mixture and stirred for 1 h at room temperature, then left overnight. The reaction mixture was diluted with methanol (0.6 ml) and purified by MDAP chromatography (method A). The desired fractions were combined, concentrated, triturated with diethyl ether, then dried under high vacuum to give the title compound. LCMS (method F): retention time=1.04, [M+H] ⁺ =594.

The following compounds were prepared in a manner analogous to that of Compound 59 using the following amines:

3-Aaminopropan-1-ol,

1-(methylamino)propan-2-ol

Compound 62 and Compound 63

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-N-(2-hydroxypropyl)-N,2-dimethylbenzamide, isomer 1 (compound 62) and isomer 2 (compound 63)

Compound 61 was purified using a Chiralpak AS-H column (30 mm × 250 mm, 5 µm) eluting with acetonitrile containing 0.2% isopropylamine to give the title compounds. Isomer 1: LCMS (method F): retention time=0.98, [M+H] ⁺ =582. Chiral HPLC: retention time 6.85, 100%. Isomer 2: LCMS (method F): retention time=0.98, [M+H] ⁺ =582. Chiral HPLC: retention time 8.36, 98.5%.

Compound 64

2-((2-methoxy-5-(2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)-5-(tetrahydro-2H-pyran-4- yl)pyrazolo[1,5-a]pyrimidin-3-yl)Phenyl)sulfonyl)ethan-1-ol

2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (18 mg, 0.086 mmol), Pd XPhos G2 was added to the dried vessel. (6 mg, 7.63 µmol), Pd / C (51 mg, 0.048 mmol), tripotassium phosphate (51 mg, 0.240 mmol) and tert-butyl (5-chloro-3- (3- ((2-hydroxyethyl) sulfonyl) - 4-methoxyphenyl)-2-methylpyrazolo[1,5-а]pyrimidin-7-yl)(3-(1-methyl-1H-imidazol-2-yl)benzyl)carbamate (intermediate 102, 55 mg, 0.082 mmol ). The vessel was closed with a lid and sparged with nitrogen. Water (0.2 ml) and 1,4-dioxane (0.8 ml) were added and the mixture was heated to 100°С for 45 min. The mixture was allowed to cool to room temp. Ammonium formate (1.25 M in methanol) (0.659 ml, 0.824 mmol) was added and the mixture was stirred overnight. The mixture was heated to 40° C. for 6 hours. The vessel was purged with nitrogen and the contents filtered through celite, washed with dichloromethane (10 ml). The filtrate was separated with water (10 ml) and the separated aqueous phase was washed with dichloromethane (2×10 ml). The combined organic layers were passed through a hydrophobic sintered filter and concentrated under reduced pressure. The residue was combined with the residue from a similar reaction carried out with tert-butyl-5-chloro-3-(3-((2-hydroxyethyl)sulfonyl)-4-methoxyphenyl)-2-methylpyrazolo[1,5-a]pyrimidine-7 -yl)(3-(1-methyl-1H-imidazol-2-yl)benzyl)carbamate (intermediate 102, 20 mg, 0.03 mmol) was dissolved in methanol (1.4 ml) and hydrochloric acid (3 M in CPME) was added ) (0.35 ml, 1.05 mmol). The mixture was stirred overnight at room temp., then at 40° C. for 7 hours. The mixture was allowed to stand at room temp. over the weekend. Saturated water was added. sodium bicarbonate (1 ml) and stirred the mixture for 5 minutes. DCM (2 ml) was added and the mixture was separated. The separated aqueous phase was washed with dichloromethane (2×1 ml) and the combined organic phases were passed through a hydrophobic sintered filter and concentrated under a stream of inert gas. The residue was dissolved in DMSO:methanol (0.5 ml, 1:1) and purified by MDAP chromatography (method A). Product-containing fractions were combined, concentrated under reduced pressure, and dried in a high vacuum apparatus to give the title compound. LCMS (method F): retention time=0.95, [M+H] ⁺ =617.

Compound 65

5-(5-(1-hydroxybuten-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-N-(3-hydroxypropyl)-N,2-dimethylbenzamide

Prepared in a manner analogous to Compound 37 using 2-(3-iodo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidin-5-yl)buten-1-ol (intermediate 104) and N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2 -dioxaborolan-2-yl)benzamide (intermediate 36) to give the title compound. LCMS (method F): retention time=0.97, [M+H] ⁺ =596.

Compounds 66 and 67

5-(5-(1-hydroxybuten-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-а] pyrimidin-3-yl)-N-(3-hydroxypropyl)-N,2-dimethylbenzamide, isomer 1 (compound 66) and isomer 2 (compound 67)

Compound 65 was purified using a Chiralpak AD-H column (30 mm × 250 mm, 5 µm) eluting with 40% ethanol (containing 0.2% isopropylamine) in heptane (containing 0.2% isopropylamine) to give the title compounds. , isomer 1: LCMS (method J): retention time=0.58, [M+H] ⁺ =596. Chiral HPLC: retention time 9.41, 100%, isomer 2: LCMS (method J): retention time=0.58, [M+H] ⁺ =596. Chiral HPLC: retention time 14.57, 99.5%.

Compound 68

5-(5-(3-hydroxypentan-3-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin3-yl)-N-(3-hydroxypropyl)-N,2-dimethylbenzamide

Prepared in a manner analogous to Compound 37 using 3-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidin-5-yl)pentan-3-ol (intermediate 105) and N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2 -dioxaborolan-2-yl)benzamide (intermediate 36) to give the title compound. LCMS (method J): retention time=0.71, [M+H] ⁺ =610.

Compound 69

N-(3-hydroxypropyl)-5-(5-(1-hydroxypropyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1, 5-а]pyrimidin-3-yl)-N,2-dimethylbenzamide, isomer 1

Prepared analogously to compound 37 using 5-yl)propan-1-ol, isomer 1 (intermediate 108) and N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2 -dioxaborolan-2-yl)benzamide (intermediate 36) to give the title compound. LCMS (method J): retention time=0.65, [M+H] ⁺ =582.

Compound 70

N-(3-hydroxypropyl)-5-(5-(1-hydroxypropyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1, 5-a]pyrimidin-3-yl)-N,2-dimethylbenzamide, isomer 2

Prepared analogously to compound 37 using 1-(3-bromo-2-methyl-7-((3-(1-methyl-1 H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidine -5-yl)propan-1-ol, isomer 2 (intermediate 109) and N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3, 2-dioxaborolan-2-yl)benzamide (intermediate 36) to give the title compound. LCMS (method J): retention time=0.61, [M+H] ⁺ =582.

Compound 71

2-((5-(5-(1-hydroxyethyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-2-methoxyphenyl)sulfonyl)ethan-1-ol, isomer 1

Prepared analogously to compound 37 using 5-yl)ethan-1-ol, isomer 1 (intermediate 111) and 2-((2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl )phenyl)sulfonyl)ethanol (intermediate 19) to give the title compound. LCMS (method F): retention time=0.87, [M+H] ⁺ =577.

Compound 72

2-((5-(5-(1-hydroxyethyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-2-methoxyPhenyl)sulfonyl)ethan-1-ol, isomer 2

Prepared analogously to compound 37 using 5-yl)ethan-1-ol, isomer 2 (intermediate 112) and 2-((2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl )phenyl)sulfonyl)ethanol (intermediate 19) to give the title compound. LCMS (method F): retention time=0.86, [M+H] ⁺ =577.

Compound 73

2-Chloro-5-(5-(1-hydroxyethyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-а] pyrimidin-3-yl)-N-(3-hydroxypropyl)-N-methylbenzamide, isomer 1

Prepared analogously to compound 37 using 5-yl)ethan-1-ol, isomer 1 (intermediate 111) and 2-chloro-N-(3-hydroxypropyl)-N-methyl-5-(4,4,5,5-tetramethyl-1,3 ,2-dioxaborolan-2-yl)benzamide (intermediate 23) to give the title compound. LCMS (method F): retention time=0.93, [M+H] ⁺ =588.

Compound 74

2-Chloro-5-(5-(1-hydroxyethyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-N-(3-hydroxypropyl)-N-methylbenzamide, isomer 2

Prepared analogously to compound 37 using 5-yl)ethan-1-ol, isomer 2 (intermediate 112) and 2-chloro-N-(3-hydroxypropyl)-N-methyl-5-(4,4,5,5-tetramethyl-1,3 ,2-dioxaborolan-2-yl)benzamide (intermediate 23) to give the title compound. LCMS (method F): retention time=0.93, [M+H] ⁺ =588.

Compound 75

1-(3-(3-((2-hydroxyethyl)sulfonyl)-4-methoxyPhenyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo [1,5-а]pyrimidin-5-yl)propan-1-ol, isomer 1

Prepared analogously to compound 37 using 5-yl)propan-1-ol, isomer 1 (intermediate 108) and 2-((2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl )phenyl)sulfonyl)ethanol (intermediate 19) to give the title compound. LCMS (method F): retention time=0.91, [M+H] ⁺ =591.

Compound 76

1-(3-(3-((2-hydroxyethyl)sulfonyl)-4-methoxyphenyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo [1,5-а]pyrimidin-5-yl)propan-1-ol, isomer 2

Prepared in a manner analogous to Compound 37 using 1-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a ]pyrimidin-5-yl)propan-1-ol, isomer 2 (intermediate 109), and 2-((2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane -2-yl)phenyl)sulfonyl)ethanol (intermediate 19) to give the title compound. LCMS (method F): retention time=0.91, [M+H] ⁺ =591.

Compound 77

5-(5-(1-hydroxypropyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-а]pyrimidin-3- yl)-2-methoxy-N-methyl-N-(tetrahydro-2H-pyran-4-yl)benzamide, isomer 1

Prepared analogously to compound 37 using 5-yl)propan-1-ol, isomer 1 (intermediate 108), and 2-methoxy-N-methyl-N-(tetrahydro-2H-pyran-4-yl)-5-(4,4,5, 5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 34) to give the title compound. LCMS (method J): retention time=0.63, [M+H] ⁺ =624.

Compound 78

5-(5-(1-hydroxypropyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-а]pyrimidin-3- yl)-2-methoxy-N-methyl-N-(tetrahydro-2H-pyran-4-yl)benzamide, isomer 2

Prepared analogously to compound 37 using 5-yl)propan-1-ol, isomer 2 (intermediate 109), and 2-methoxy-N-methyl-N-(tetrahydro-2H-pyran-4-yl)-5-(4,4,5, 5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 34) to give the title compound. LCMS (method J): retention time=0.64, [M+H] ⁺ =624.

Compound 79

5-(5-(1-hydroxycyclopropyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-3- yl)-N-(3-hydroxypropyl)-N,2-dimethylbenzamide

tert-Butyl (3-bromo-5-(1-((tert-butyldimethylsilyl)oxy)cyclopropyl)-2-methylpyrazolo[1,5-a]pyrimidin-7-yl)(3-(1-methyl-1H- imidazol-2-yl)benzyl)carbamate (intermediate 115, 26 mg, 0.039 mmol), potassium phosphate (24 mg, 0.113 mmol), XPhos (2 mg, 4.20 µmol), XPhos Pd G2 (3 mg, 3.81 µmol) and N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 36, 25 mg, 0.053 mmol) were combined in 1,4-dioxane (0.4 ml) and water (0.133 ml). The mixture was heated to 60° C. in a sealed vessel for 1 hour 45 minutes. XPhos Pd G2 (3 mg, 3.81 µmol) and N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl ) benzamide (intermediate 36, 25 mg, 0.053 mmol). After 1 h, the reaction temperature was raised to 80°C. After 2 h XPhos Pd G2 (3 mg, 3.81 µmol) and N-(3-hydroxypropyl)-N,2-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane- 2-yl)benzamide (intermediate 36, 25 mg, 0.053 mmol) and heated the mixture to 100°C for 1 h. The mixture was partitioned between water (10 ml) and ethyl acetate (10 ml). The separated aqueous phase was washed with ethyl acetate (2×10 ml) and the combined organic phases were passed through a hydrophobic sintered filter and concentrated under reduced pressure. The residue was dissolved in 1,4-dioxane (1.0 ml) and then HCl (4M in 1,4-dioxane) (0.1 ml) was added. The mixture was stirred at room temperature for 4 days. Saturated aqueous sodium bicarbonate solution (3 ml) was added and the mixture was stirred for 10 minutes. Water (10 ml) and DCM (10 ml) were added and the phases were separated. The separated aqueous phase was washed with dichloromethane (2×5 ml) and the combined organic layers were passed through a hydrophobic sintered filter and concentrated under reduced pressure. The residue was purified by MDAP chromatography (method A). Fractions containing the desired product were combined and concentrated under reduced pressure to give the title compound. LCMS (method F): retention time=0.97, [M+H] ⁺ =580.

Compound 80

N-((S)-1-hydroxypropan-2-yl)-5-(5-(1-hydroxypropyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl) benzyl)amino)pyrazolo[1,5-a]pyrimidin3-yl)-N,2-dimethylbenzamide, isomer 1

Prepared analogously to compound 37 using 5-yl)propan-1-ol, isomer 1 (intermediate 108), and (S)-N-(1-hydroxypropan-2-yl)-N,2-dimethyl-5-(4,4,5, 5-tertramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 45) to give the title compound. LCMS (method J): retention time=0.62, [M+H] ⁺ =582.

Compound 81

N-((S)-1-hydroxypropan-2-yl)-5-(5-(1-hydroxypropyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl) benzyl)amino)pyrazolo[1,5-a]pyrimidin3-yl)-N,2-dimethylbenzamide, isomer 2

Prepared analogously to compound 37 using 5-yl)propan-1-ol, isomer 2 (intermediate 109), and (5)-N-(1-hydroxypropan-2-yl)-N,2-dimethyl-5-(4,4,5, 5-tertramethyl-1,3,2-dioxaborolan-2-yl)benzamide (intermediate 45) to give the title compound. LCMS (method J): retention time=0.61, [M+H] ⁺ =582.

Compound 82

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-N-(2-hydroxypropyl)-2-methoxy-N-methylbenzamide

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-2-methoxybenzoic acid (intermediate 120, 65 mg, 0.101 mmol), HATU (47 mg, 0.124 mmol) and DIPEA (0.055 ml, 0.315 mmol) were stirred in THF (1 ml) at room temperature for 5 min, after which 1-(methylamino)propan-2-ol (13 mg, 0.146 mmol) was added. After 2 h more HATU (12 mg, 0.032 mmol) was added and DIPEA (0.018 ml, 0.101 mmol) was added. After 1.5 h, the reaction mixture was purified by MDAP chromatography (method A). Product-containing fractions were combined and dried under a stream of inert gas to give the title compound. LCMS (method F): retention time=0.96, [M+H] ⁺ =598.

Compounds 83 and 84

5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a] pyrimidin-3-yl)-N-(2-hydroxypropyl)-2-methoxy-N-methylbenzamide, isomer 1 (compound 83), and isomer 2 (compound 84)

Compound 82 was purified on a Chiralpak IC column (30 mm × 250 mm, 5 µm) eluting with 30% ethanol (containing 0.2% isopropylamine) in heptane (containing 0.2% isopropylamine) to give the title compounds, isomer 1 : LCMS (method F): retention time=0.94, [M+H] ⁺ =598. Chiral HPLC: retention time 28.8, 100%. Isomer 2: LCMS (method F): retention time=0.94, [M+H] ⁺ =598. Chiral HPLC: retention time 33.68, 96.6%.

Compound 85

2-((2-methoxy-5-(2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)-5-((tetrahydro-2H-pyran-4 -yl)methyl)pyrazolo[1,5-a]pyrimidin-3-yl)phenyl)sulfonyl)ethan-1-ol

Prepared analogously to compound 9 using 3-iodo-2-methyl-N-(3-(1-methyl-1H-imidazol-2-yl)benzyl)-5-((tetrahydro-2H-pyran-4-yl)methyl )pyrazolo[1,5-a]pyrimidine-7-amine (intermediate 122) and 2-((2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2 -yl)phenyl)sulfonyl)ethanol (intermediate 19) to give the title compound. LCMS (method F): retention time=0.94, [M+H] ⁺ =631. Compound 86

2-(3-(3-((2-hydroxyethyl)sulfonyl)-4-methoxyphenyl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo [1,5-a]pyrimidin-5-yl)buten-1-ol

Prepared analogously to compound 37 using 5-yl)buten-1-ol (intermediate 104) and 2-((2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl) sulfonyl)ethanol (intermediate 19) to give the title compound. LCMS (method F): retention time=0.92, [M+H] ⁺ =605.

Compound 87

3-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-а] pyrimidin-3-yl)-N-(3-hydroxypropyl)-N-methylbenzamide

2-(3-bromo-2-methyl-7-((3-(1-methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-5-yl)propan- 2-ol (intermediate 43, 150 mg, 0.313 mmol), potassium phosphate (165 mg, 0.777 mmol), dicyclohexyl(2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl )phosphane (12 mg, 0.025 mmol), XPhos Pd G2 (22 mg, 0.028 mmol), and N-(3-hydroxypropyl)-N-methyl-3-(4,4,5,5-tetramethyl-1,3, 2-dioxaborolan-2-yl)benzamide (intermediate 123, 219 mg, 0.412 mmol) was combined in a microwave vial. 1,4-dioxane (1.8 ml) and water (0.6 ml) were added, the vessel was sealed, and the mixture was heated to 100°C in a microwave reactor for 1 h. After cooling, the organic layer of the biphasic reaction mixture was filtered through cotton wool and directly purified by MDAP chromatography (method A). The resulting residue was partitioned between saturated aqueous sodium bicarbonate (10 ml) and DCM (5 ml). The separated aqueous phase was washed with additional DCM (2×5 ml). The organics were combined, passed through a hydrophobic sintered filter, concentrated under a stream of inert gas, and dried under high vacuum at 40° C. for 1 day to give the title compound. LCMS (method F): retention time=0.94, [M+H] ⁺ =568.

Biological analyzes

a) Analysis of PI4KB activity

Inhibition of human PI4KIII-beta 13-828 (D316-330) activity in vitro was determined using the ADP-Glo Kinase assay from Promega. The inhibitors were dissolved in 100% DMSO at a concentration of 1 mM. Dilutions in 100% DMSO were obtained by 1/3 step serial dilutions. A 60 nl sample from an 11-point titration was transferred to a small volume white 384-well Greiner assay plate, achieving a final DMSO concentration of 1% in the whole plate and a highest final inhibitor concentration of 10 μM. PI4KIII-beta assay included 25 mM Hepes pH 7.5 (NaOH), 10 mM MgCl2, 0.5 mM EGTA, 0.1% Triton X-100, 2 mM TCEP, and 0.1 mg/mL BSA, 1 mM ATP, 60 μM phosphatidylinositol and 1.5 nM human PI4K-beta 13-828 (D316-330) in a total volume of 6 μl. The beginning of the analysis was initiated by adding the enzyme, covered and incubated at room temperature for 3 hours. The assays were stopped by adding 6 μl of ADP-GLO reagent containing 0.1% CHAPS to deplete unconsumed ATP. The plates were incubated at room temperature for 60 minutes, and then 12 μl of kinase detection reagent containing 0.1% CHAPS was added to convert ADP to ATP, and luciferase and luciferin were injected. After 40 minutes of incubation at room temperature, the luminescent signal was read on a BMG Labtech PHERAstar FS using the following settings; gain 3600, focus height 13.7 mm, measurement interval 1 s, settling time 0.2 s. The inhibitory effect of compounds on PI4K-beta activity was assessed by IC50 response versus high control, no compound, and low control, no enzyme, by fitting a four-parameter dose-response equation.

When tested in this assay: all compounds produced a mean pIC50 greater than or equal to 8.1; compounds 1-7, 9-11, 14, 15, 17-20, 22-29 and 31-87 gave a mean pIC50 greater than or equal to 8.4; compounds 17 and 19 gave a mean pIC50 of 8.6; compound 21 gave a mean pIC50 of 8.2; and Compound 22 gave a mean pIC50 of 8.5.

b) Retention time analysis

Caliper preparatory analysis

Compound dilutions were made in 100% DMSO by 1/3 step serial dilutions at the highest concentration of 1 mM. A 200 nL sample from a 12-point titration was transferred to a black 384-well Greiner assay plate, achieving a final concentration of 1% DMSO in the entire plate and a final inhibitor concentration of 10 μM in a final assay volume of 20 μl. A well of a 384-well assay plate was pre-incubated with 10 μl of 2X enzyme buffer with compound for 1 hour. Buffer without enzyme was used as a control with 100% inhibition. 10 μl of 2X substrate solution was added to initiate the start of the assay, so that the final concentrations were 2 mM ATP and 1 μM Bodipy-PI, and the plate was incubated at 25°C for 3 hours. The enzymatic reaction was stopped by adding 40 μl of stop buffer; the substrate (PI) and product (PIP) present in each sample were separated electrophoretically using a LabChip 3000 capillary electrophoresis instrument, CALIPER LABCHIP 3000 Drug Discovery System, and detected using a blue laser (480 nm) for excitation and CCD in the green range (520 nm) for detection (CCD2). Negative control samples (0% inhibition in the absence of inhibitor) and positive control samples (100% inhibition in the absence of enzyme) were collected in 48 replicates (4 replicates per Caliper sipper) and used to calculate the percentage inhibition in each test well. Percent inhibition (Pinh) was determined using the following equation:

Pinh=(PSR0% - PSRinh)/(PSR0% - PSR100%) × 100

where PSRinh is the ratio of product to total in the presence of inhibitor, PSR0% is the average ratio of product to total in the absence of inhibitor, and PSR100% is the average ratio of product to total in control samples with 100% inhibition; IC50 values for inhibitors are determined by fitting inhibition curves (Pinh versus inhibitor concentration) with a 4-parameter sigmoidal dose-response model using XLfit 4 (IBDS) software.

Dilution with rapid measurement of inhibitor concentration In a small Eppendorf tube, 5 µl of 2x enzyme solution (0.4 µM - 0.8 µM) was mixed with 5 µl of 2x compound solution with 40-120-fold higher IC50 values determined with 2 mM ATP as described above and incubated for 1 hour at room temperature. The start of the assay was initiated by rapidly mixing 2 μl of the pre-incubated enzyme mixture with 800 μl of the substrate mixture containing 2 mM ATP and 1 μM Bodipy-PI. 60 μl of each sample was transferred to a small volume 384-well Grainer plate; the substrate (PI) and product (PIP) present in each sample were separated electrophoretically using a LabChip 3000 capillary electrophoresis instrument, CALIPER LABCHIP 3000 Drug Discovery System, and detected using a blue laser (480 nm) for excitation and CCD in the green range (520 nm) for detection (CCD2) for approximately 8 hours (200 measurements per well). Process curves showing % conversion vs. time in the presence of the compound are compared with curves for control samples without compound. The curves were fitted with the process curve equation using the XLfit software:

where Vi is the initial rate of the enzyme, Vs is the rate of operation at equilibrium in the presence of the diluted inhibitor, and t is the time after dilution.

The observed values of Vi, Vs and Kobs depend on the nature of the inhibitor (ie fast equilibration or strong binding). Depending on the initial conditions of the analysis (the relative concentration of the compound and the enzyme), the Vi and Vs values can be preset and/or fixed. For slowly dissociating compounds > [E], the initial rate of the enzyme (after rapid dilution) is usually not fixed; the rate of operation at steady state can be fixed at the rate in the presence of residual compound (determined from the IC50 curve). For some fast equilibrating compounds tested (1/2 retention time<resolution of assay) > [E], the initial work rate can be fixed at the theoretical % activity level determined from the IC50 curves at the respective compound concentrations, and the steady state work rate can be pre-set at the control level, but not fixed. For other fast equilibrating or slowly dissociating compounds, an alternative method of pre-calculating Vi and Vs not fixed at the theoretical rates determined from IC50 curves at appropriate compound concentrations can be used, for example, in cases where the above strategies do not give satisfactory results. approximations.

In this assay, compound 17 showed an average retention time of 542 minutes; compound 19 showed an average retention time of 997 minutes; compound 21 showed an average retention time of 528 minutes; and compound 22 showed an average retention time of 1666 minutes.

c) Cytopathic effect analysis protocol (CPE)

Inhibition of PI4KB activity in vitro was determined by ATP depletion inhibition assay in HeLa Ohio cells in response to infection with human rhinovirus type A strain 16 (HRVA16). ATP levels were determined using the CellTiter-GLO reagent from Promega. Compounds that inhibit PI4KB are also potent inhibitors of human rhinoviruses and are able to protect HeLa Ohio cells from CPE induced by viral infection and replication. Protected cells remain viable after viral infection and, accordingly, their ATP levels are higher. Compounds were dissolved in 100% DMSO to a concentration of 3 mM followed by a 1:3 stepwise serial dilution in 100% DMSO. A 0.5 µl sample from a 10-point titration was transferred to a white 96-well flat-bottomed Grainer tissue culture plate (655083). A 0.5 µl sample of 100% DMSO was added to plate rows 11 and 12, resulting in a total plate DMSO concentration of 0.33% and the highest total compound concentration in the assay of 10 µM.

HeLa Ohio cells were cultured at 37°C, 5% CO2 in medium (DMEM supplemented with 10% Australian fetal bovine serum and 2 mM glutamax). Cells were passaged after reaching >80% confluency. For analysis, cells were cultured to 80-90% confluency until dissociated.

Cells were dissociated for analysis by washing in PBS and dissociating using 3 ml TrypLE Express for 5 ml at 37°C. Dissociated cells were mixed with 7 ml of medium and centrifuged at 300 g for 5 minutes. The cell pellet was resuspended in 50 ml of medium and the cells were counted using a Beckman Coulter ViCell. The cells were diluted to a density of 6.6×104 cells/ml and some of the cell volume was transferred to a new tube to be used as a 12 row control. HRVA16 stock material was added to the remaining cell suspension using an appropriate dilution of viral stock material to achieve an MOI of 1.

150 µl of cell and virus suspension was added to each well of vertical rows 1-11 using the Multidrop Combi. 150 μl of a separate cell suspension was added to each well in a 12 vertical row using a multichannel pipette. The assay plates were sealed and incubated at 33°C, 5% CO2 for two days. After 2 days, the assay plates were removed from the incubator and left to equilibrate to room temperature. 60 µl CellTiter-Glo reagent was added to all wells using the Thermo Scientific Multidrop Combi. The plates were incubated at room temperature for 20 minutes and then read on the Perkin Elmer Envision (settings: capture height 2.5, fixed measurement height (mm) 6.5, distance between plate and detector 0, measurement time (s) 0, 1, luminescence correction factor (ct2) (0%)).

The inhibitory effect of compounds on PI4KB activity was assessed by IC50 response relative to high (no virus, vertical row 12) and low (virus + no inhibitor, vertical row 11) controls by fitting a four-parameter dose-response equation.

When tested in this assay: all compounds gave a mean pIC50 equal to or greater than 7.2; compounds 1-8, 10-15 and 17-87 gave a mean pIC50 greater than or equal to 7.6; compounds 1, 3, 5, 6, 7, 10-12, 14, 15, 17-25, 29, 31, 33-37, 39-53, 55-59, 61-68, 70, 76, 80-82 , 84 and 86 gave a mean pIC50 greater than or equal to 8.4; compounds 17 and 19 gave a mean pIC50 of 8.9; compound 21 gave a mean pIC50 of 8.8; and Compound 22 gave a mean pIC50 of at least 9.1.

d) Metabolic stability analysis in human microsomes

Brief description of the protocol

Test compound (0.5 μM) was incubated with pooled liver microsomes. The test compound was incubated for a 45 minute experiment and analyzed by LC-MS/MS.

Experimental Procedure Pooled human liver microsomes were purchased from a reliable commercial supplier such as Corning Life Sciences. Microsomes (final protein concentration 0.5 mg/mL), 50 mM phosphate buffer pH7.4 and NADPH (final concentration=1 mM) were pre-incubated at 37°C until test compound was added (final substrate concentration=0.5 μM; final DMSO concentration=0.25%) to initiate the reaction. The final incubated volume was 500 µl. For each compound tested, a control incubation was included with 50 mM phosphate buffer, pH 7.4, instead of NADPH (minus NADPH) being added. Two control compounds for each species were included. All incubations are carried out individually for each test compound.

Each compound was incubated for 45 minutes and incubated samples (50 μl) were taken at time points at 0, 5, 15, 30 and 45 minutes. A control sample (minus NADPH) was taken at 45 minutes only. Reactions were stopped by adding 100 μl of acetonitrile containing an internal standard to the sample. After termination of the reaction, the samples were centrifuged at 2500 rpm for 20 minutes at 4°C to precipitate the protein.

Quantitative Analysis

After protein precipitation, the samples were analyzed by LC-MS/MS under standard conditions.

Data analysis

From a plot of the peak area ratio (compound peak area/internal standard peak area) versus time, the slope of the curve was determined. The intrinsic clearance was then calculated using the equations below and then converted to ml/min/g:

Elimination rate constant (k)=(- slope)

Half-life (t1/2) (min)=0.693+k

Intrinsic clearance (CLint) (µl/min/mg protein)=(V×0.693)+t _1/2

where V=Incubation volume, µl/mg of microsomal protein.

When tested in this assay, intrinsic clearance of compound 17 was 28.9 ml/min/g; intrinsic clearance of compound 19 was 11.6 ml/min/g; intrinsic clearance of compound 21 was 34.0 ml/min/g; and intrinsic clearance of compound 22 was 33.7 ml/min/g.

f) Spleen concentration analysis

Compounds 17, 18, 19, 22 and 23 were tested in this spleen accumulation assay. Compounds were administered intravenously to rats to determine the level of the compound in the spleen upon complete absorption of the absorbed dose (orally or through the lungs) and the introduction into the systemic circulation of the full dose.

Compounds 17, 19, 22 and 23 were formulated in 2% DMSO in kleptose (aq., 10% w/v). Compound 18 was formulated with DMSO:PEC200:water in a ratio of 5:45:50. The formulations were administered as intravenous infusions (1 mg/kg over 1 hour) to one male Wistar Han rat.

12 hours after the start of dosing, rats were sacrificed and spleens were collected. Samples of spleens weighing approximately 0.5 g were taken and homogenized in 4 ml of water. 100 µl of final homogenates were obtained by protein precipitation with 300 µl of acetonitrile containing an internal standard and analyzed (along with calibration standards prepared in the same way) using reverse phase liquid chromatography-mass spectrometry using a heated electrospray source and monitoring multiple reactions in positive ion mode. As a column for liquid chromatography, Waters Cortecs C18 with a particle size of 2.7 μm, a column size of 50×2.1 mm, at 60°C was used; (A) 0.1% formic acid (aq) and (B) 0.1% formic acid in acetonitrile were used as mobile phase, which functioned as a gradient, while operating at a flow rate of 1 ml/min.

The concentration of all compounds in the spleen was less than 30 ng/g. The concentration of compounds 17, 19, 22 and 23 in the spleen was less than 7 ng/g. In particular, the concentration of compound 17 in the spleen was 1.76 ng/g, and the concentration of compound 22 in the spleen was 5.54 ng/g.

Claims (9)

1. The compound which is (S)-N-(1-hydroxypropan-2-yl)-5-(5-(2-hydroxypropan-2-yl)-2-methyl-7-((3-(1 -methyl-1H-imidazol-2-yl)benzyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)-N,2-dimethylbenzamide (compound 22)

or a pharmaceutically acceptable salt thereof. 3. A compound according to claim 1 or a pharmaceutically acceptable salt thereof for use in the treatment of a rhinovirus infection. 4. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, for use in the treatment of a disorder caused or exacerbated by a rhinovirus infection. 5. A compound according to claim 1 or a pharmaceutically acceptable salt thereof for use according to claim 4, characterized in that the disorder is chronic obstructive pulmonary disease (COPD). 6. A method for treating a rhinovirus infection, comprising administering a compound according to claim 1 or a pharmaceutically acceptable salt thereof to a patient in need thereof. 7. A method of treating a disorder caused or aggravated by a rhinovirus infection, comprising administering a compound of claim 1 or a pharmaceutically acceptable salt thereof to a patient in need thereof. 8. Pharmaceutical composition for the treatment of a rhinovirus infection or a disorder caused or aggravated by a rhinovirus infection, containing an effective amount of a compound according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.