Heterocyclic derivatives as janus kinase inhibitors

Abstract

This invention relates to compounds of general formula (I) cyclic carbamates and derivatives of ureabenzimorpholine and benzothiomorpholine that inhibit JAK family non-receptor tyrosine protein kinases (JAK1, JAK2, JAK3, and TYK2); methods for preparing such compounds; pharmaceutical compositions containing them; and their therapeutic uses. The compounds of this invention can be used to treat diseases or conditions associated with dysregulation of JAK family non-receptor kinases; particularly for the treatment of various inflammatory diseases, including asthma, COPD, and other respiratory diseases.

Description

Technical Field

[0001] This invention relates to chemical compounds that can be used as JAK inhibitors, such as oxadiazolidine and oxadiazolidine benzomorpholine and benzothiomorpholine derivatives of JAK 1, which can be used to treat a variety of inflammatory diseases, including asthma, COPD and other respiratory diseases. Background Technology

[0002] The JAK family consists of non-receptor tyrosine protein kinases and has four major members: JAK1, JAK2, JAK3, and TYK2. More than 50 cytokines and growth factors bind non-covalently to type I and type II receptors of JAK kinases in various combinations. Ligand-triggered signaling involves tyrosine phosphorylation of the receptor by JAK and recruitment of one or more STAT proteins. Tyrosine-phosphorylated STATs dimerize and are then transported across the nuclear membrane to the nucleus to regulate specific genes. JAKs possess seven homologous domains (JAK homologous domains, JH). Starting from the carboxyl terminus, JH1 is the first JH, called the kinase domain, and consists of approximately 250 amino acid residues. JH1 encodes the kinase protein that constitutes the kinase domain of the phosphorylated substrate; JH2 is the pseudokinase domain that regulates the activity of the kinase domain. JAK3 is expressed in the bone marrow and lymphatic system, as well as in endothelial cells and vascular smooth muscle cells; other members are expressed in almost all tissues (Hu X et al., Signal Transduct Target Ther. 2021, 26; 6(1):402). Many cellular processes are downstream of JAK / STAT signaling: hematopoiesis, immune homeostasis, tissue repair, inflammation, apoptosis, and adipogenesis. Different biological responses are regulated by specific pairings of JAK isotypes. The JAK1 / JAK3 combination mediates IL-2, -4, -7, -9, -15, and -21 signaling, which are associated with lymphocyte growth / maturation, T cell / NK cell differentiation / homeostasis, B cell class switching, and other inflammatory processes. The JAK1 / TYK2-JAK1 / JAK2 combination regulates signals associated with innate immune responses, such as IL-6 and type I interferon, which are involved in naive T cell differentiation, T cell homeostasis, granulocyte production, and other inflammatory processes. (Howell MD et al., Front. Immunol. 2019, 10, 2342). JAK2 is often associated with itself (JAK2 / JAK2) to control the signaling of various cytokines and growth factors, such as IL-3, IL-5, granulocyte-macrophage colony-stimulating factor (GM-CSF), erythropoietin (EPO) and thrombopoietin (TPO) (Hodge et al., Clin Exp Rheumatol 2016; 34(2):318-28).

[0003] Genetically modified mouse models and human diseases have demonstrated the importance of the JAK / STAT pathway in immune adaptation. In particular, overexpression or mutation of certain JAK isotypes and aberrant JAK / STAT signaling drive malignancies in hematopoietic and lymphoid tissues, as well as inflammatory conditions. Currently, several JAK inhibitors approved by the Food and Drug Administration (FDA) and / or the European Union are in clinical use. Two small molecules (ruxolitinib and fedratinib) are used for hematologic disorders such as myelofibrosis and polycythemia vera; six JAK inhibitors (tofacitinib, baricitinib, ruxololitinib, filgotinib, upadicitinib, and delgocitinib, in Japan) are used for immune-mediated disorders such as rheumatoid arthritis, polyarticular juvenile idiopathic arthritis, atopic dermatitis, ulcerative colitis, and acute graft-versus-host disease. Furthermore, some of these drugs, along with others, are currently undergoing Phase II and Phase III clinical trials to cover indications ranging from autoimmune diseases (lupus, vitiligo, etc.), inflammatory bowel disease to non-Hodgkin's lymphoma and COVID-19 (Hu X. et al., Sig Transduct Target Ther 2021, 6:402).

[0004] Small molecules targeting JAK / STAT represent an attractive option for treating fibrotic conditions. Indeed, inflammatory cytokines (IL-4, IL-3, IL-6, IL-11, IL-31, etc.) and growth factors (FGF, VEGF, etc.) involved in the fibrotic process activate the JAK / STAT pathway. Ruxolitinib, tested in a bleomycin-induced fibrosis mouse model, improved fibrotic lesions in the lungs and reduced levels of molecular markers of fibrosis (Zhang, Y et al., Ann. Rheum. Dis. 2017, 76, 1467-1475), while tofacitinib acted as a prophylactic agent in experimental cutaneous and pulmonary fibrosis (Wang, W et al., Scleroderma Relat. Disord. 2020, 5, 40-50). Several case reports have been investigated in patients. Six case reports confirmed the efficacy and safety of tofacitinib combined with nintedanib in managing aggressive interstitial lung disease with poor prognosis (Conca, W et al., Front. Pharmacol. 2020, 11, 5857619). Baricitinib has been shown to be a safe immunomodulator that reduces the levels of biomarkers of pulmonary fibrosis and inflammation in patients with RA, including a subgroup with interstitial lung disease (D'Alessandro M et al., Int. Immunopharmacol. 2020, 86, 106748).

[0005] Several JAK inhibitors are currently undergoing clinical trials for COVID-19: tofacitinib, baricitinib, and ruxolitinib. Baricitinib and ruxolitinib have been associated with a reduced risk of death. They have reduced the use of invasive mechanical ventilation and have a marginal effect on intensive care unit admissions and the incidence of acute respiratory distress syndrome (ARDS). (Wijaya, I. et al. Clin. Epidemiol. Glob. Health 2021, 11, 100755). Ruxolitinib has also been tested in COVID-19 patients, improving clinical symptoms and chest computed tomography images (Cao Y. et al. J. Allergy Clin. Immunol. 2020 146, 137-146).

[0006] Asthma can be included among a wide range of immune-mediated diseases, and its pathogenesis is characterized by the important role of JAK / STAT signaling. Asthma is a chronic inflammatory disease of the airways, resulting from a complex interplay between immune responses, genetic susceptibility, and nonspecific external stimuli such as cold, allergens, and exercise, leading to airway hyperresponsiveness, remodeling, and ultimately airflow limitation. Severe asthma affects 5% to 15% of the adult asthma population (300 million people worldwide) and represents a public health problem associated with increased mortality, increased hospitalizations, a significant symptom burden, healthcare costs, and missed work and school (Steve NG et al., J Allergy Clin Immunol 2021; 148:953-63). Severe asthma represents a subset of difficult-to-treat asthma and occurs in patients whose disease remains uncontrolled despite the use of high-dose inhaled corticosteroids (ICS) in combination with long-acting beta-agonists or other controller agents. To date, four types of biologics have been licensed for use in severe asthma: omalizumab (anti-immunoglobulin E), mepolizumab and reslizumab (anti-interleukin [IL]-5), benralizumab (anti-IL-5 receptor α), and dupilumab (anti-IL-4 receptor α). Despite their efficacy, many patients continue to experience exacerbations or uncontrolled disease, indicating a need for more novel therapies (Israel E, Reddel HK. N Engl J Med 2017; 377:965-76).

[0007] Recent advancements in the understanding of asthma pathobiology have led to a shift from phenotypic classification systems to the introduction of the concept of “endotypes.” The latter classifies asthma based on pathophysiological mechanisms and clinical biomarkers relevant to a given patient (Wenzel SE et al., Am J Respir Crit Care Med 2021; 203:809-21). Two main endotypes exist in asthma: type 2 and non-type 2. The type 2 pathway is defined by the activation of cytokines derived from TH2 cells and group 2 innate lymphocytes (ILC2); these include IL-4, IL-5, and IL-13, which induce airway inflammation by activating eosinophils, B cells, airway epithelial cells, and other cell types. Biomarkers of type 2 asthma include hematologic / sputum eosinophilia and elevated fractional levels of exhaled nitric oxide (FENO) and IgE. Type 2-low pathway is characterized by the absence of type 2-high cytokines and biomarkers, and it exhibits increased or oligogranulocytic neutrophil levels in the airways, with normal levels of airway neutrophils and eosinophils. Type 2-low asthma is not well understood and may encompass a variety of different endotypes. Potential mediators and / or biomarkers of T2-low endotype under investigation include IL-6, IL-17A / F, IL-23, type I interferon, CXCL10, TNF, alarm factors (TSLP, IL-25, IL-33), IL-1β, IL-8, IFN-γ (Hinks TSC et al., ERJ 2021, 57(1) 2000528).

[0008] Almost all of the aforementioned mediators of T2 and T2-low endotypes activate the JAK / STAT pathway, which is the fundamental principle behind the potential use of JAK inhibitors in both endotypes of severe asthma. The simultaneous targeting of several cytokines by JAK inhibitors may offer advantages over biologics (for unresponsive patients) and standard therapy (for patients still uncontrolled), given their administration on top of infusions (ICS).

[0009] Although JAK inhibitors have a strong theoretical basis in asthma, safety concerns may arise due to the administration of systemic depressants, or may limit their use in certain asthma subjects, such as children. Given that asthma is a lung-limiting disease, the inhaled route of administration of JAK inhibitors can offer advantages in therapeutic efficacy while limiting systemic exposure and associated side effects. To date, several companies are developing inhaled JAK inhibitors for the treatment of asthma. Astrazeneca's pipeline includes AZD-0449 (completed Phase I clinical trial) and AZD-4604 (currently in Phase I clinical trial); Theravance Biopharma is initiating a new preclinical program for the inhaled JAK inhibitor TD-8236; and Kinaset / Vectura is developing VR588 (currently in Phase I clinical trial) as an inhaled compound. Numerous preclinical studies funded by these companies have demonstrated the efficacy of JAK inhibitors in modulating asthma. In the preclinical phase of drug development, the orally administered JAK1 / 3 inhibitor R256 (now known as AZD0449) showed efficacy in reducing airway resistance, BAL eosinophilia, and mucus production, and also effectively reduced TH2 cytokine responses when administered during sensitization (Ashino S et al., J Allergy Clin Immunol 2014; 133:1162-74). iJak-381, administered as a dry powder from Genentech, reduced BAL eosinophilia, CCL11, airway resistance, and Muc5AC in mice challenged with OVA. Furthermore, it reduced BAL eosinophilia, neutrophilia, CCL11, and CXCL1 in a mouse model of chronic exposure to AAH allergens (Dengler HS et al., Sci Transl Med 2018; 10:eaao2151). In addition, formulations of oral JAK inhibitors such as tofacitinib for aerosol administration reduced eosinophil counts in a mouse model of house dust mite asthma (Younis US et al., AAPS PharmSci-Tech 2019; 20:167).

[0010] Another respiratory disease that can benefit from lung-limiting JAK suppression is chronic obstructive pulmonary disease (COPD), an inflammatory disease of the lungs most commonly caused by exposure to cigarette smoke, characterized by largely irreversible and progressive airflow limitation. Although inflammatory cytokines are drivers of chronic airway inflammation, and some of them trigger JAK / STAT activation (IL-6, IFN-γ, IL-2, etc.), the role of this pathway in the pathogenesis of COPD is poorly characterized. Phosphorylated STAT4+ cells were found to be increased in COPD compared to healthy non-smokers (Di Stefano A et al., Eur Respir J. July 2004; 24(1):78-85). In another study, lung biopsies from COPD patients showed higher counts of phosphorylated STAT3+ and phosphorylated STAT1+ cells than those from non-smokers, although it was impossible to reproduce previous data on phosphorylated STAT4 molecules (Yew-Booth L et al., Eur Respir J 2015; 46(3):843-5). These data may also suggest the therapeutic potential of JAK inhibitors in COPD.

[0011] Although administered by inhalation, safety concerns may arise due to the reaching of drug levels throughout the system after inhalation of JAKi. In addition to its suitability for inhalation, JAKi should preferably possess additional properties that further limit systemic exposure after inhalation.

[0012] There remains a strong need for JAK inhibitors, particularly inhaled JAK inhibitors, which have the potential to provide compounds with improved safety.

[0013] WO2022 / 194781A1 discloses a structurally similar compound as a JAK inhibitor, which differs from this application at least in terms of the pattern of substituents.

[0014] Given the number of pathological reactions mediated by JAK enzymes, there is a continued need for JAK enzyme inhibitors that can be used to treat many conditions, especially respiratory diseases.

[0015] Therefore, the discovery of novel, safe, and effective JAK inhibitors suitable for local application to the lungs to treat asthma and respiratory diseases remains an important need. Summary of the Invention

[0016] Therefore, one object of the present invention is to provide compounds of formula (I) benzomorpholine and benzothiomorpholine.

[0017]

[0018] R1, R2, and R3 are as defined in the detailed description of this invention; or pharmaceutically acceptable salts thereof, which can be used as JAK kinase inhibitors.

[0019] Another object of the present invention is to provide a pharmaceutical composition comprising such a compound, a method of treating respiratory diseases using such a compound, and methods and intermediates for preparing such a compound.

[0020] In one aspect, the present invention provides a compound of formula (I) for use as a medicament. In another aspect, the present invention provides the use of the compounds of the present invention in the preparation of medicaments.

[0021] In another aspect, the present invention provides the use of the compounds of the present invention for the preparation of medicaments for treating any disease related to the JAK enzyme mechanism.

[0022] In another aspect, the present invention provides a method for preventing and / or treating any disease related to the JAK enzyme mechanism as defined above, the method comprising administering a therapeutically effective amount of the compound of the present invention to a patient requiring such treatment.

[0023] In one specific aspect, the compounds of the present invention may be used alone or in combination with other active ingredients and may be administered for the prevention and / or treatment of lung diseases, including asthma, chronic obstructive pulmonary disease (COPD), cystic fibrosis (CF), interstitial lung disease and idiopathic pulmonary fibrosis (IPF), acute lung injury and acute respiratory distress syndrome (ARDS). Detailed Implementation

[0024] definition

[0025] The term “pharmaceutically acceptable salt” refers to a derivative of a compound of formula (I), wherein the parent compound is appropriately modified by converting any free acid or basic group (if present) into the corresponding addition salt using any base or acid that is conventionally expected to be pharmaceutically acceptable.

[0026] Therefore, suitable examples of the salt may include inorganic or organic acid addition salts of basic residues such as amino groups, and inorganic or organic base addition salts of acid residues such as carboxyl groups.

[0027] The inorganic base cations suitable for preparing the salts of the present invention include ions of alkali metals or alkaline earth metals such as potassium, sodium, calcium, or magnesium. Salts containing, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, camphorsulfonic acid, acetic acid, oxalic acid, maleic acid, fumaric acid, succinic acid, and citric acid are obtained by reacting the main alkali-acting compound with an inorganic or organic acid to form a salt.

[0028] Many organic compounds can form complexes with solvents in which they react or from which they precipitate or crystallize. These complexes are called "solvents," which is another object of the present invention. Polymorphs and crystalline forms of compounds of formula (I) or pharmaceutically acceptable salts or solvates thereof are another object of the present invention.

[0029] The term "halogen," "halogenated," or "halogen atom" includes fluorine, chlorine, bromine, and iodine atoms; it refers to fluorine, chlorine, bromine, and iodine as substituents.

[0030] The term "(C1-C6)alkyl" refers to a straight-chain or branched alkyl group having 1 to 6 carbon atoms. Specific alkyl groups include, for example, methyl, ethyl, n-propyl, isopropyl, tert-butyl, 3-methylbutyl, etc.

[0031] The term "(C1-C6)alkyl" refers to a straight-chain or branched alkyl group having 1 to 6 carbon atoms. Specific alkyl groups include, for example, methyl, ethyl, n-propyl, isopropyl, tert-butyl, 3-methylbutyl, etc.

[0032] The term "(C1-C6)haloalkyl" refers to the "(C1-C6)alkyl" group as defined above, in which one or more hydrogen atoms are replaced by one or more halogen atoms that may be the same as or different from each other. Examples include halogenated, polyhalogenated, and fully halogenated alkyl groups, in which all hydrogen atoms are replaced by halogen atoms, such as trifluoromethyl or difluoromethyl groups.

[0033] By analogy, the term "(C1-C x )hydroxyalkyl" or "(C1-C x "(C1-C1)aminoalkyl" refers to "(C1-C1)aminoalkyl" as defined above. x The term "(C1-C6)hydroxyalkyl" refers to a hydroxyalkyl or aminoalkyl group in which one or more hydrogen atoms are replaced by one or more hydroxyl (OH) or amino groups, and where x is an integer of up to 10. Therefore, "(C1-C6)hydroxyalkyl" or "(C1-C6)aminoalkyl" refers to the hydroxyalkyl or aminoalkyl group in which the number of carbon atoms is in the range of 1 to 6.

[0034] The definition of aminoalkyl encompasses alkyl groups substituted with one or more amino groups (-NR4R5) (i.e., "(C1-C6)alkyl" groups). Examples of aminoalkyl groups are mono-aminoalkyl groups, such as R4R5N-(C1-C6)alkyl or -(CH2). m NR4R5, where R4 and R5 and m are as defined in the detailed description of the present invention.

[0035] Regarding the substituents R4 and R5 as defined above, further explanation is provided below: when R4 and R5 together with the nitrogen atom to which they are attached form a 5- to 6-membered heterocyclic group, at least one additional ring carbon atom in the heterocyclic group may be replaced by at least one heteroatom or heterogroup (e.g., N, NH, S, or O) or may be substituented with an oxonium substituent (=O). The heterocyclic group may also optionally be substituted at a usable site in the ring, i.e., on a carbon atom, or on a heteroatom or heterogroup that can be substituted. Therefore, examples of such heterocyclic groups are 1-pyrrolidinyl, 1-piperidinyl, 1-piperazinyl, 4-morpholinyl, piperazine-4-yl-2-one, and 4-methylpiperazin-1-yl.

[0036] The term "(C3-C)" 10 "(C3-C6)cycloalkyl" refers to a saturated cyclic hydrocarbon group containing the indicated number of carbon atoms in the ring. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl, as well as polycyclic systems such as adamantyl.

[0037] The term "aryl" refers to a monocyclic, bicyclic, or tricyclic carbocyclic system having 6 to 20, preferably 6 to 15, ring atoms, wherein at least one ring is aromatic. Examples of suitable aryl ring systems include, for example, phenyl or naphthyl, indenyl, or dihydroindenyl groups.

[0038] The term "heteroaryl" refers to a monocyclic, bicyclic, or tricyclic system having 5 to 20, preferably 5 to 15, ring atoms, wherein at least one ring is aromatic and at least one ring atom is a heteroatom (e.g., N, NH, S, or O). Examples of suitable heteroaryl ring systems include, for example, thiophene, pyrrole, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, pyridinyl, pyrazinyl, pyridazinyl, triazinyl, furanylpurine, indole, isoyindole, indazole, etc.

[0039] The derived expression "(C3-C 10 "(C3-C6) heterocyclic alkyl" also refers to a saturated or partially unsaturated monocyclic alkyl, bicyclic alkyl, or tricyclic alkyl group with the indicated number of carbon atoms, wherein at least one ring atom is a heteroatom (e.g., N, NH, S, or O) or may additionally have an oxo substituent (=O) (e.g., C(=O), S(=O)2).

[0040] The group may be optionally substituted, wherein the term “optionally substituted” means substituted or unsubstituted. When the term “one or more” refers to any atom or group that is a substituent of a group of a compound of formula (I), it is intended that there are 1 to 3, preferably 1 to 2, more preferably 1 such substituent that may replace hydrogen on such a group or variable.

[0041] Substitution includes bridging systems, spirodisubstituted, and substitution on two adjacent atoms, in both cases thus forming additional 5- to 6-membered heterocycles. Examples of (C3-C6) heterocyclic alkyl groups are represented by the following: oxobutyryl, tetrahydrofuranyl, pyrrolidinyl, imidazoalkyl, thiazoalkyl, piperazinyl, piperidinyl, morpholinyl, thiomorpholinyl, 9-methyl-3,9-diazaspiro[5.5]undecane-3-yl, and (3aR,6aS)-5-methyl-octahydropyrrolo[3,4-c]pyrrolidin-2-yl.

[0042] The term "aryl (C1-C6) alkyl" refers to an aryl ring attached to a straight-chain or branched alkyl group, wherein the number of carbon atoms in the ring ranges from 1 to 6, such as phenylmethyl (i.e., benzyl), phenylethyl, or phenylpropyl.

[0043] Similarly, the term "heteroaryl (C1-C6)alkyl" refers to a heteroaryl ring attached to a straight-chain or branched alkyl group, wherein the number of constituent carbon atoms is in the range of 1 to 6, such as furanylmethyl.

[0044] The term "alkylyl" refers to an HC(O)- or alkyl carbonyl group (e.g., (C1-C6)alkylC(O)-), where the group "alkyl" has the meaning as defined above. Examples include formyl, acetyl, propionyl, and butyryl.

[0045] The term "(C1-C)" 10 )alkoxy" or "(C1-C 10 "(C1-C6)alkoxy", "(C1-C6)alkoxy", or "(C1-C6)alkoxy" refer to straight-chain or branched hydrocarbons of the indicated number of carbon atoms connected to the rest of the molecule via oxygen bridges. "(C1-C6)alkylthio" refers to the aforementioned hydrocarbons connected via sulfur bridges.

[0046] The derived terms "(C1-C6)haloalkoxy" or "(C1-C6)haloalkoxy" refer to a haloalkyl group as defined above, linked by an oxygen bridge. Examples of (C1-C6)haloalkoxy groups are difluoromethoxy and trifluoromethoxy.

[0047] Similarly, the derived expressions “(C3-C6)heterocyclic alkyl-(C1-C6)alkyl” and “(C3-C6)cycloalkyl-(C1-C6)alkyl” refer to heterocyclic alkyl and cycloalkyl groups as defined above, connected to the rest of the molecule by an alkyl group with the indicated number of carbon atoms, such as piperidin-4-yl-methyl and cyclohexylethyl.

[0048] The derived expression “(C1-C6)alkoxy(C1-C6)alkyl” refers to an alkoxy group as defined above, such as methoxymethyl, connected to the rest of the molecule by an alkyl group of the indicated carbon number.

[0049] Similarly, "(C1-C6)haloalkoxy(C1-C6)alkyl" refers to a "(C1-C6)haloalkoxy" group as defined above, which is attached to the rest of the molecule by an alkyl group of the indicated number of carbon atoms, such as difluoromethoxypropyl.

[0050] Similarly, "(C1-C6)alkoxycarbonyl" refers to an alkoxy group as defined above that is attached to the rest of the molecule via a carbonyl group.

[0051] "(C1-C6)alkylthiocarbonyl-" refers to the aforementioned alkylthio group that is attached to the rest of the molecule via a carbonyl group (C=O).

[0052] "(C1-C6)alkoxycarbonyl-(C1-C6)alkyl" refers to an alkoxy group as defined above, which is connected to the rest of the molecule by a carbonyl group further connected to an alkyl group of the indicated number of carbon atoms, such as methoxycarbonylmethyl.

[0053] "(C1-C6)alkoxycarbonyl-(C1-C6)alkylthio" therefore refers to the connecting group such as methoxycarbonylmethylthio.

[0054] Other derived expressions will be self-evident in their meaning.

[0055] For example, "halogenated-((C1-C6)alkyl(C3-C8)heterocyclic alkyl)" refers to the linked group such as 4-fluoro-1-methylpyrrolidine-3-yl.

[0056] The oxygen subunit is represented by (O), as an alternative to other common representations such as (=O). Therefore, carbonyl groups are preferably represented herein as -C(O)-, as an alternative to other common representations such as -CO-, -(CO)-, or -C(=O)-. Generally, groups enclosed in parentheses are side groups not included in the chain, and parentheses are used to help disambiguate linear chemical formulas when deemed useful; for example, sulfonyl-SO2- can also be represented as -S(O)2- to dispel ambiguity, for example, regarding sulfinyl-S(O)O-.

[0057] When labeled with a number, the statement (value) "p is zero" or "p is 0" means that the substituent or group with the label p (e.g., Ip) is not present, that is, there are no substituents other than H when needed. Similarly, when the label is attached to a bridging divalent group (e.g., (CH2)m), the statement "m is zero each time it appears..." or "m is 0" means that the bridging group is not present, that is, it is a bond.

[0058] Keys pointing to wavy or curved lines, such as the keys used in the structural formulas in this article. It describes the bond that serves as the connection point between a part or substituent and the core or main chain structure.

[0059] The term "bond" used to define a substituent refers to the situation where two functional groups connected by a substituent are directly linked to each other without any other atoms in between.

[0060] A dash ("-") not between two letters or symbols is intended to indicate the connection point of a substituent.

[0061] Whenever a basic amino or quaternary ammonium group is present in a compound of formula (I), a physiologically acceptable anion may be present, selected from chloride, bromide, iodide, trifluoroacetate, formate, sulfate, phosphate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, tartrate, oxalate, succinate, benzoate, p-toluenesulfonate, dihydroxynaphthylate, and naphthalenedisulfonate. Similarly, in the presence of an acidic group such as a COOH group, a corresponding physiologically acceptable cation salt may also be present, including alkali metal or alkaline earth metal ions.

[0062] Compounds of formula (I) can exist as optical stereoisomers when they contain one or more stereoisomer source centers.

[0063] When the compounds of the present invention have at least one stereoisomer source center, they can exist as enantiomers accordingly. When the compounds of the present invention have two or more stereoisomer source centers, they can also exist as diastereomers. It should be understood that all such single enantiomers, diastereomers, and mixtures thereof in any proportion are covered within the scope of the present invention. The absolute configuration (R) or (S) of the carbon with the stereoisomer source center is specified based on the priority of the group according to the Cahn-Ingold-Prelog nomenclature rule.

[0064] When reported near the chemical name of a compound, “single stereoisomer,” “single diastereomer,” or “single enantiomer” indicates that the isomer has been isolated as a single diastereomer or enantiomer (e.g., by chiral chromatography), but the absolute configuration at the center of the relevant stereoisomer source is not determined / specified.

[0065] The hindered rotation isomers are generated by hindered rotation around a single bond, where the spatial strain barrier of the rotation is high enough to allow the separation of conformational isomers (Bringmann G et al., Angew. Chem. Int. Ed. 44 (34), 5384-5427, 2005. doi:10.1002 / anie.200462661).

[0066] Oki defined a resisted isomer as a conformational isomer that interconverts with a half-life of more than 1,000 seconds at a given temperature (Oki M, Topics in Stereochemistry 14, 1-82, 1983).

[0067] Unlike other chiral compounds, hindered transisomers can reach thermal equilibrium in many cases, whereas in other forms of chiral isomerization they are usually only chemically possible.

[0068] The separation of trans-restricted isomers is possible through chiral resolution methods such as selective crystallization. In trans-enantioselective or trans-restriction-selective synthesis, the formation of one trans-restricted isomer comes at the expense of another. Trans-restriction-selective synthesis can be carried out using chiral auxiliaries such as Corey Bakshi Shibata (CBS) catalysts (asymmetric catalysts derived from proline) or via thermodynamic equilibrium-based pathways (where the isomerization reaction favors one trans-restricted isomer over another).

[0069] The racemic forms of compounds of formula (I), as well as individual sterically isomers (which are substantially free of their corresponding enantiomers) and mixtures of sterically isomer-rich sterically isomers, are included within the scope of this invention.

[0070] The present invention also relates to corresponding deuterated derivatives of compounds of formula (I). In the context of the present invention, a deuterated derivative means that at least one position occupied by a hydrogen atom is occupied by a amount of deuterium higher than its natural abundance. Preferably, the percentage of deuterium at this position is at least 90%, more preferably at least 95%, and even more preferably 99%.

[0071] All preferred groups or embodiments of the compounds of formula (Io) described above and below can be combined with each other, and necessary modifications also apply.

[0072] The term "IC" 50 "The half maximum inhibitory concentration" refers to the half maximum inhibitory concentration, which is a measure of the efficacy of a substance in inhibiting a specific biological or biochemical function.

[0073] The term "pIC" 50 "IC" refers to IC50 expressed as molar concentration. 50 The negative logarithm of the value.

[0074] As described above, the present invention provides compounds of general formula (I) that act as JAK inhibitors, and methods for preparing such compounds comprising pharmaceutical compositions, either alone or in combination with one or more pharmaceutically acceptable carriers, mixed with one or more pharmaceutically acceptable carriers.

[0075] Therefore, in one aspect, the present invention relates to compounds of general formula (I):

[0076]

[0077] in

[0078] R1 is selected from the following heteroaryl groups.

[0079] ;

[0080] R2 is selected from

[0081] and

[0082] in

[0083] K is selected from O and S;

[0084] R3 is a monocyclic urea or carbamate group selected from the following formula J.

[0085]

[0086] R7 is selected from H, (C1-C6)alkyl, (C1-C6)hydroxyalkyl, and -(CH2). m NR4R5, (C1-C6)alkoxycarbonyl (CH2) m ,

[0087] Where m is independently 0 or an integer from 1 to 4 each time it appears;

[0088] R4 and R5 may be the same or different, and are selected independently.

[0089] -H、

[0090] (C1-C6)alkyl,

[0091] (C1-C6)alkoxy-(C1-C6)alkyl;

[0092] Its single enantiomers, diastereomers, and mixtures,

[0093] Or its pharmaceutically acceptable salts or solvates.

[0094] All listed groups in each of the variable portions R1, R2, R3, R4, R5, R7, J1, J2 of the compounds of the present invention must be intended as selections and may be combined with each other in embodiments included within the scope of the present invention.

[0095] In a preferred embodiment, the present invention relates to compounds of formula (I) as Jak inhibitors, wherein

[0096] R1 is pyrazolo[1,5-a]pyrimidin-3-yl, R2 is

[0097]

[0098] Where K is O;

[0099] Represented by general formula (Ib)

[0100]

[0101] in

[0102] R3 is J2; and

[0103] R7 is selected from H, (C1-C6) alkyl, and -(CH2). m NR4R5,

[0104] And its pharmaceutically acceptable salts and solvates.

[0105] In another preferred embodiment, the present invention relates to compounds of formula (I) as Jak inhibitors, wherein

[0106] R1 is (3-oxomylidene-3,4-dihydropyrazin-2-yl)amino.

[0107] R2 is

[0108]

[0109] in

[0110] K is 0;

[0111] Represented by the general formula (Ic)

[0112]

[0113] in

[0114] R3 is J2; and

[0115] R7 is selected from H, (C1-C6) alkyl, and -(CH2). m NR4R5,

[0116] Or its pharmaceutically acceptable salts or solvates.

[0117] In another preferred embodiment, the present invention relates to at least one of the compounds listed in Table 1 below, and pharmaceutically acceptable salts thereof.

[0118] Table 1 - List of preferred compounds

[0119]

[0120]

[0121]

[0122] The compounds of the present invention exhibit high biochemical potency against JAK targets (JAK1, JAK2, JAK3 and Tyk2) and high potency in representative cellular functional assays (e.g., inhibition of pSTAT6 in BEAS cells stimulated with IL-13).

[0123] Lung retention is a complex interaction between solubility, permeability and lung protein binding. The preferred compounds of the present invention have favorable physicochemical properties that can be translated into good inhalation characteristics.

[0124] The compounds of the present invention, including all those listed above, can be prepared from readily available starting materials using the general methods and procedures described in the Experimental Section below, or by using slightly modified methods readily available to those skilled in the art. Although specific embodiments of the invention may be shown or described herein, those skilled in the art will recognize that all embodiments or aspects of the invention can be prepared using the methods described herein or by using other known methods, reagents, and starting materials. Other method conditions may also be used unless otherwise stated, when typical or preferred method conditions (i.e., reaction temperature, time, molar ratio of reactants, solvent, pressure, etc.) are given. While optimal reaction conditions can vary depending on the specific reactants or solvents used, such conditions can be readily determined by those skilled in the art through conventional optimization methods. The preparation methods described below and reported in the following embodiments should not be considered as limiting the scope of synthetic methods that can be used to prepare the compounds of the present invention.

[0125] In some cases, steps are required to mask or protect sensitive or reactive parts, and according to general principles of chemistry (Protective group in organic syntheses, 3rd edition, TWGreene, PGMWuts), commonly known protecting groups (PGs) can be used.

[0126] For clarity, the compounds of formula (I) reported here again include all the compounds listed in Table 1 above, which can generally be prepared according to the procedures shown in the following scheme. When specific details or steps differ from the general scheme, they have been described in detail in specific examples and / or other schemes.

[0127]

[0128] Compounds of formula (I) may contain one or more stereoisomeric source centers. Enantiomerically pure compounds can be prepared by commonly known reactions, such as those described below, using enantiomerically pure starting materials and intermediates. These intermediates are commercially available or readily produced by those skilled in the art from commercial sources.

[0129] In another method, enantiomerically pure compounds can be prepared by means of chiral chromatography from the corresponding racemic mixtures and / or partially racemic mixtures.

[0130] Compound (I) can be prepared according to Scheme 1. Compound II is an intermediate that can be deprotected by the protecting groups of PG1 and / or PG2 to convert it into a compound of Formula I. Obviously, in the absence of PG1 and PG2, any general method described for preparing intermediate II will provide a compound of general formula (I).

[0131] The suitable PG1 protecting group of the secondary NH of the 6-methoxy-3,4-dihydro-2H-benzo[b][1,4]oxazine-7-yl moiety included in the r2 of intermediate II (and intermediate IV) can be a carbamate-type protecting group, such as Boc (tert-butoxycarbonyl). The Boc-protected intermediate II can be readily removed by treating it with a strong organic or inorganic acid under acidic conditions. For example, the Boc group can be cleaved by treating the intermediate with pure or mixed trifluoroacetic acid, typically overnight at room temperature, either directly or in a mixture with organic solvents such as DCM, DCE, THF, etc.

[0132] Intermediate II can be directly introduced into R1 (or R1 appropriately protected with PG2, named r1) through metal / palladium-catalyzed cross-coupling reactions such as Suzuki coupling, Stille coupling, Buchwald-Hartwig et al. (Strategic application of named reactions in organic synthesis, edited by L. Kurti and B. Czako, 2005), and obtained through the reaction of intermediate IV and intermediate III.

[0133] For example, when R1 is pyrazolo[1,5-a]pyrimidin-3-yl, a suitable palladium-catalyzed cross-coupling for introducing R1 is Suzuki coupling. Suzuki coupling can be achieved by using a Pd catalyst such as tetrakis(triphenylphosphine)palladium(0), PdCl2(dppf)2, or a ligand-palladium ring precatalyst such as XPhos-Pd-G3. The reaction is carried out in the presence of [(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate] in an organic solvent such as 1,4-dioxane, THF, 1,2-dimethoxyethane, 2-propanol or DMF, with or without water, in the presence of an inorganic base such as an alkali metal carbonate (e.g., Cs2CO3 or K2CO3) or an inorganic phosphate (e.g., K3PO4), under heating (typically 50-100 °C) for several hours (typically 1 to 3 h), causing intermediate IV to react with the corresponding boric acid or pinacol ester of borate (intermediate III, wherein R1 is pyrazolo[1,5-a]pyrimidin-3-yl, A is dihydroxyboryl or 4,4,5,5-tetramethyl-1,3,2-dioxaborane). Boric acid and pinacol esters of borate are generally commercially available, or can be readily prepared by those skilled in the art from commercially available reagents.

[0134] When R1 is (3-oxoylidene-3,4-dihydropyrazin-2-yl)amino, the suitable palladium-catalyzed cross-coupling for introducing R1 is the Buchwald-Hartwig coupling. For synthetic convenience, the carbonyl group of the (3-oxoylidene-3,4-dihydropyrazin-2-yl)amino group needs to be masked with PG2, for example, by using an alkoxy group such as a methoxy group. Intermediates IV and III (where r1 is 3-methoxypyrazin-2-amino and A is H) can be synthesized in suitable ligand palladium ring systems such as XPhos-Pd-G3 (2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate) or RuPhos-Pd-G3. (2-Dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate) or a generally suitable Pd source (e.g., Pd2(dba)3 or Pd(OAc)2) is reacted with a suitable biphenylphosphine ligand type (RuPhos, X-Phos or similar) in the presence of a strong organic base such as sodium tert-butoxide or an inorganic base such as Cs2CO3 in an organic solvent such as 1,4-dioxane, THF or toluene at a high temperature (typically 80-120 °C) for several hours (typically 1-5 h).

[0135] When PG2 is present, the removal of the methoxy group can be carried out by treating intermediate II with TMS-Cl (trimethylchlorosilane) and sodium iodide in acetonitrile at 60-100 °C for 1 to 5 h. Under these reaction conditions, PG1 can also be removed simultaneously without additional treatment.

[0136]

[0137] Intermediate IV can be prepared according to Scheme 2. Intermediate IV can be prepared by an N-arylation of intermediate V with halide intermediate r2-Lg via a copper-catalyzed Ullmann-type reaction. The Ullmann reaction between NH heteroaryl and aryl / heteroaryl halides (bromines or iodides) can be carried out in the presence of a suitable copper (I) catalyst / co-catalyst such as CuI, Cu2O or CuTC (copper thiopheneformate), without ligands or with suitable ligands such as N,N-dimethylglycine, proline, phenanthroline, dimethylcyclohexane-1,2-diamine (DMCHA), in the presence of an inorganic base such as K2CO3 or Cs2CO3, by heating (typically 90-150°C) overnight or longer in a polar organic solvent such as DMSO, DMF or DMA.

[0138] Intermediate V can be prepared from intermediate VIa in a two-step process involving: 1) CN coupling of VIa with R3-H and 2) deprotection of PG3. The first step of the reaction of VIa with R3-H can be carried out under Ullmann conditions by heating (typically 60-100 °C) in an organic solvent such as DMSO, in the presence of a copper (I) catalyst / co-catalyst such as CuI, Cu2O, or CuTC (copper thiophenecarboxylate), without ligands or with suitable ligands such as proline, 3,4,7,8-tetramethyl-1,10-phenanthroline, N,N-dimethylglycine, dimethylcyclohexane-1,2-diamine (DMCHA), and an inorganic base such as K2CO3, K3PO4, or Cs2CO3 with heterocyclic iodides and heterocyclic amines. When PG3 is THP (tetrahydropyranyl), the deprotection of PG3 can be carried out by acidic treatment with an organic acid such as TFA at room temperature for several hours (typically 12-24 hours) in the presence of a scavenger such as a silyl hydride such as triethylsilane.

[0139]

[0140] In various methods, intermediate IV, where r2 / R2 is 6-methoxy-2H-benzo[b][1,4]thiazine-3(4H)-one-7-yl, can be interconverted via functional groups, such as reducing a lactam to an amine to an intermediate of formula IV, where r2 / R2 is 6-methoxy-3,4-dihydro-2H-benzo[b][1,4]thiazine-7-yl. The reduction of a lactam can be carried out by reacting it with a suitable reducing agent, such as borane, in a suitable organic solvent, such as THF, at a temperature from 0°C to room temperature.

[0141] In another method, when R3 is J1, intermediate IV (named IVa) can be prepared in a three-step process starting from intermediate VIIb according to scheme 3. This three-step process includes: 1) carbamylation of the amino derivative VIIb with isocyanate VIIIa, 2) cyclization to form a cyclic urea, and 3) alkylation of the secondary nitrogen with R7-X. The carbamylation reaction can be carried out by reacting the amine derivative and the corresponding isocyanate in an organic solvent such as DCM or THF in the presence of an organic base such as DIPEA or TEA, typically overnight at room temperature. Subsequently, step 2 can be carried out by treating the carbamylated derivative of step 1 in an organic solvent such as THF or dioxane with a strong base such as potassium tert-butoxide, sodium ethoxide, or an alkali metal hydride, typically overnight at room temperature. In step 3, the alkylation of NH and R7-X of the cyclic urea can be carried out by reacting the cyclic urea and a suitable halide (typically X=Br) in an organic solvent such as THF or dioxane in the presence of a strong base such as potassium tert-butoxide, sodium ethoxide or an alkali metal hydride, typically overnight at room temperature.

[0142]

[0143] In another method, when R3 is J2 and R7 is H, intermediate IV (named IVb) can be prepared in a two-step process starting from intermediate VIIb according to scheme 3. This two-step process includes: 1) carbamylation of chloroformate VIIIb with amino derivative VIIb, followed by 2) cyclization to form an oxazolidinone. The carbamylation reaction can be carried out by reacting the amine derivative and a suitable chloroformate in an organic solvent such as DCM or THF in the presence of an organic base such as pyridine or DIPEA, typically at temperatures ranging from 0°C to room temperature for several hours (typically 1-3 h). Subsequently, step 2 can be carried out by treating the carbamylated derivative of step 1 in an organic solvent such as DMF or dioxane with an organic base such as morpholine or piperidine, and heating at a high temperature (typically 70-90°C) for several hours (typically 1-3 h).

[0144] In different methods, when R3 is J2, intermediate IV (named IVc) can be prepared from intermediate VIId according to scheme 3. The two-step, one-pot reaction of ethylene oxide IX with intermediate VIId can be carried out first by treating intermediate VIId for a short time (typically 1 h) in an organic solvent such as THF or dioxane at a low temperature (typically 0 °C) and in the presence of a strong base such as LiHDMS or LDA (lithium diisopropylamino), followed by reaction with intermediate IX at a higher temperature (typically 90–120 °C) for several hours (typically 12–24 h) to obtain intermediate IVc'. In some cases, the one-pot method produces an intermediate amino alcohol, which can be readily converted to the corresponding oxazolidinone by treatment overnight in toluene with diethyl carbonate at a high temperature (typically at reflux temperature).

[0145] When R7 is an aminoalkyl derivative such as -(CH2) m In NR4R5, intermediate IVc can be derived from a precursor with r7 as appropriate protection, such as -(CH2), according to scheme 4. m The intermediate IVc′ (see Scheme 3) (named IVc″) of N-PG4 is prepared, wherein PG4 is a divalent orthogonal protecting group such as phthalimide. Intermediate VIc″ is used to prepare intermediate IVc (where R7 is -(CH2)). m The conversion of NR4R5 can be carried out in a two-step process, which involves deprotection of PG4 followed by insertion of R4 / R5 via reductive amination and / or alkylation. Deprotection of PG4 can be performed by treating the corresponding VIc″ at room temperature for several hours (typically 1 to 3 hours) in an alcohol solvent such as EtOH or iPrOH in the presence of hydrazine (typically hydrazine hydrate) to provide the corresponding primary amine Vic″ (where r7 is -(CH2)). m NH2). IVc (where R7 is -(CH2)). m The preparation of NR4R5 (where R4=R5=Me) can be carried out by reducing the amination intermediate VIc'' with formaldehyde in methanol or ethanol in the presence of acetic acid and a reducing agent such as NaB(CN)H3 or Na(OAc)3H. The preparation of IVc (where R7 is -(CH2)) m NR4R5) where R4 is Me but R5 is a different group, can be achieved by first monoalkylating IVc''' with a suitable alkylating agent such as R5-Br, followed by reductive amination. Alkylation can be carried out in the presence of an organic base such as DIPEA or TEA in an organic solvent such as DMF or dioxane by heating (typically about 60°C) to react IVc''' with the halide R5-Br for several hours (usually overnight), yielding the intermediate IVc (where R7 is -(CH2)). mNHR5) can be converted into intermediate IVc (where R7 is -(CH2)) by reductive amination following the same procedure described above. m NR4R5), where R4 is Me and R5 is a different group.

[0146]

[0147] The preparation of intermediates VIIb, VIIc, and VIId used in Scheme 3 is reported in Scheme 5. Intermediates VIb (and VIc) can be converted to intermediates VIIb (and VIc) under Ullmann conditions via a CN coupling reaction with r2-Lg (where Lg is Br or I), as previously described for the conversion of VIa to V in Scheme 2. Intermediate VIId can be prepared from intermediate VIIc by curtious degradation in a one-pot two-step process, which includes: firstly, reacting an acid with a suitable coupling agent such as propanephosphonic anhydride and a suitable azide source such as TMS-N3 in the presence of an organic base such as TEA or DIPEA in a suitable organic solvent such as 2-Me-THF, THF, or dioxane at reflux temperature for 1 h or longer to convert acid VIIc to the corresponding isocyanate; in the second step, the formed isocyanate can be reacted with the corresponding alcohol such as benzyl alcohol in a one-pot reaction at reflux temperature for several hours (typically 12-24 h) to give the corresponding carbamate derivative VIId.

[0148]

[0149] The above scheme can provide at least one non-limiting synthetic route for preparing Examples 1 to 14.

[0150] As described in detail herein, the compounds of the present invention are inhibitors of kinase activity, particularly JAK kinase activity, for the treatment of JAK-dependent diseases.

[0151] In one aspect, the present invention provides compounds according to the invention, namely compounds of formula (I) or pharmaceutical compositions thereof, which are used as medicines, preferably for the prevention and / or treatment of respiratory diseases, particularly lung diseases.

[0152] In another aspect, the present invention provides the use of compound (I) or a pharmaceutically acceptable salt thereof in the preparation of medicaments for treating conditions related to the JAK mechanism, particularly for treating conditions such as respiratory diseases and lung diseases.

[0153] In particular, the present invention provides compounds of formula (I) for the prevention and / or treatment of lung diseases selected from asthma, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), acute lung injury, and acute respiratory distress syndrome (ARDS).

[0154] Furthermore, the present invention provides a method for preventing and / or treating conditions related to the JAK mechanism, the method comprising administering a therapeutically effective amount of the compound of the present invention to a patient requiring such treatment.

[0155] In particular, the present invention provides methods for prevention and / or treatment of respiratory diseases selected from asthma, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), acute lung injury, and acute respiratory distress syndrome (ARDS).

[0156] Preferably, the compound of the present invention is used for the prevention of the aforementioned conditions.

[0157] Also preferred is the use of the compounds of the present invention for treating the aforementioned conditions.

[0158] Generally speaking, compounds that act as JAK inhibitors can be used to treat many conditions related to the JAK enzyme mechanism.

[0159] In one embodiment, the conditions that can be treated by the compounds of the present invention are selected from asthma, chronic obstructive pulmonary disease (COPD), and interstitial lung diseases such as idiopathic pulmonary fibrosis (IPF), acute lung injury, and acute respiratory distress syndrome (ARDS).

[0160] In another implementation, the condition is selected from asthma and chronic obstructive pulmonary disease (COPD).

[0161] The treatment methods of the present invention comprise administering to a patient in need an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. As used herein, an "effective amount" of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or other pharmaceutically active agent, means an amount sufficient to treat the patient's condition but low enough to avoid serious side effects, and can still be routinely determined by those skilled in the art. A compound of formula (I) or a pharmaceutically acceptable salt thereof may be administered once or according to a dosing regimen in which multiple doses are administered at different time intervals over a given time period. Typical daily doses may vary depending on the particular route of administration chosen.

[0162] The present invention also provides pharmaceutical compositions of compounds of formula (I) mixed with one or more pharmaceutically acceptable carriers or excipients, such as those described in Remington's Pharmaceutical Sciences Handbook, 17th edition, MackPub., NY, USA.

[0163] The present invention also relates to the use of the compounds of the present invention and pharmaceutical compositions thereof for use in various routes of administration.

[0164] The compounds of the present invention and their pharmaceutical compositions may be administered according to the patient’s needs, for example, orally, nasally, parenterally (subcutaneously, intravenously, intramuscularly, intrasternally, and by infusion), by inhalation, rectal, vaginal, topical, local, transdermal, and ocular administration.

[0165] Various solid oral dosage forms can be used to administer the compounds of the present invention, including solid forms such as tablets, capsule-shaped tablets, capsules, pouches, granules, lozenges, and bulk powders. The compounds of the present invention can be administered alone or in combination with various pharmaceutically acceptable carriers, diluents (such as sucrose, mannitol, lactose, and starch), and known excipients, including suspending agents, solubilizers, buffers, binders, disintegrants, preservatives, colorants, flavoring agents, lubricants, etc. Timed-release capsules, tablets, and gels are also advantageous.

[0166] Various liquid oral dosage forms can also be used to administer the compounds of the present invention, including aqueous and non-aqueous solutions, emulsions, suspensions, syrups, and elixirs. Such dosage forms may also contain suitable known inert diluents such as water and suitable known excipients such as preservatives, wetting agents, sweeteners, flavoring agents, etc., as well as agents for emulsifying and / or suspending the compounds of the present invention. The compounds of the present invention can be formulated into injectable compositions as isotonic sterile solutions, for example, for intravenous injection. Other formulations are also possible.

[0167] Suppositories of the compounds of the present invention for rectal administration can be prepared by mixing the compounds with suitable excipients such as cocoa butter, salicylates / esters and polyethylene glycol.

[0168] Preparations for vaginal application may be in the form of creams, gels, pastes, foams or sprays, and in addition to the active ingredient, may also contain a suitable carrier.

[0169] For topical application, the pharmaceutical composition may be in the form of creams, ointments, liniments, lotions, emulsions, suspensions, gels, solutions, pastes, powders, sprays, and drops suitable for application to the skin, eyes, ears, or nose. Topical application may also include transdermal application, such as via transdermal patches.

[0170] For the treatment of respiratory diseases, as described above, the compounds according to the present invention can also preferably be administered by inhalation.

[0171] Some preferred compounds of the present invention exhibit properties suitable for administration via inhalation.

[0172] Optimizing medicaments for inhalation delivery requires certain characteristics that allow the compound to maintain a sufficient local concentration (pulmonary retention) upon administration to the lungs to exert its pharmacological effect for the desired duration. For the swallowed portion, drug absorption in the GI tract is minimal and typically occurs at irrelevant levels in unwanted compartments (i.e., plasma). For this purpose, one or more characteristics of the compound, such as, but not limited to, membrane permeability, dissolution rate, and basicity, are optimized to enhance its binding to phospholipid-rich lung tissue or lysosomal capture. In some embodiments, the compounds of the present invention exhibit one or more of the aforementioned characteristics within the desired range for inhaled compounds.

[0173] Inhalable formulations include inhalable powders, metered aerosols containing propellants, or inhalable formulations without propellants, and can be administered by a suitable inhalation device, which may be selected from dry powder inhalers, pressurized metered inhalers, or nebulizers.

[0174] For administration as dry powder, single-dose or multi-dose inhalers known from the prior art can be used. In this case, the powder can be filled in gelatin, plastic, or other capsules, cartridges, or blister packs or storage containers.

[0175] Diluents or carriers, such as lactose or any other additives suitable for improving the breathable fraction, may be added to the powdered compounds of the present invention.

[0176] Inhalation aerosols containing propellant gases such as hydrofluorocarbons may contain the compounds of the present invention in solution or dispersion form. Propellant-driven formulations may also contain other components such as cosolvents, stabilizers, and optional other excipients.

[0177] Propellant-free inhalable formulations containing the compounds of the present invention may be in the form of solutions or suspensions in aqueous, alcoholic, or hydroalcoholic media, and may be delivered by jet or ultrasonic nebulizers known in the art or by soft mist nebulizers, such as Respimat®, a registered trademark of Boehringer Ingelheim Pharmaceuticals (Wachtel, H., Kattenbeck, S., Dunne, S. et al., Pulm Ther (2017) 3:19).

[0178] Regardless of the route of administration, the compounds of the present invention can be administered as a single active agent or in combination with other pharmaceutical active ingredients (i.e., as a co-therapeutic agent administered in fixed doses or in combination therapy of separately formulated active ingredients).

[0179] The compounds of the present invention can be administered as the sole active agent or in combination with other pharmaceutical active ingredients, including those currently used to treat respiratory disorders and known to those skilled in the art, such as β2-agonists, antimuscarinic drugs, corticosteroids, mitogen-activated protein kinase (P38 MAP kinase) inhibitors, PI3K inhibitors (phosphoinositol 3-kinase), nuclear factor κ-B kinase subunit β inhibitors (IKK2), Rho kinase inhibitors (ROCKi), human neutrophil elastase (HNE) inhibitors, phosphodiesterase 4 (PDE4) inhibitors, leukotriene modifiers, nonsteroidal anti-inflammatory drugs (NSAIDs), and mucus modifiers.

[0180] The present invention also relates to kit products comprising a pharmaceutical composition of a single compound of the present invention or a compound of the present invention in combination with or mixed with one or more pharmaceutically acceptable carriers and / or excipients, and a device that may be a single-dose or multi-dose dry powder inhaler, a metered-dose inhaler, or a nebulizer.

[0181] The dosage of the compounds of the present invention depends on a variety of factors, including the specific disease to be treated, the severity of symptoms, the route of administration, the frequency of dose intervals, the specific compound used, the efficacy of the compound, its toxicological properties, and its pharmacokinetic properties.

[0182] Pharmaceutical compositions comprising the compounds of the present invention suitable for inhalation administration are in various inhalable forms, such as inhalable powders (DPIs), metered-dose inhalers (PMDIs) containing propellants, or inhalable formulations without propellants (e.g., UDVs).

[0183] The present invention also relates to devices comprising pharmaceutical compositions containing compounds of the present invention, which may be single-dose or multi-dose dry powder inhalers, metered-dose inhalers, and nebulizers, particularly soft fog nebulizers.

[0184] The present invention is illustrated in more detail in the following embodiments.

[0185] The features of the invention will become apparent in the following description of exemplary embodiments, which are given to illustrate the invention and not to limit it.

[0186] Preparation and Examples of Intermediates

[0187] General Experimental Details

[0188] The chemical names of compounds are generated using Structure To Name Enterprise 10.0 Cambridge software or the latest version.

[0189] Purification by rapid chromatography refers to purification using a Biotage SP1 or Interchim puriFlash purification system or equivalent MPLC with a pre-packed polypropylene column containing a stationary phase (column). If Si column purification is used, this refers to an Interchim pre-packed polypropylene column containing unbound activated silica with spherical particles having an average size of 15 μm; or an Isolute® pre-packed polypropylene column containing unbound activated silica with irregular particles having an average size of 50 μm. Fractions containing the desired product (identified by TLC and / or LCMS analysis) are combined and concentrated under vacuum. Purification by reversed-phase chromatography refers to purification using a Biotage Isolera Four purification system equipped with a Biotage Dalton 2000 mass detector, at Sfar C 18 Purification on a pre-packed polypropylene column. When using an SCX-2 column, "SCX-2 column" refers to a Bond Elut® pre-packed polypropylene column containing a strong cation exchange adsorbent, uncapped propylsulfonic acid functionalized silica. In the case of purification using preparative HPLC-MDAP (MDAP: Mass-Directed Automated Purification), fractions containing the desired product are combined, and the solvent is removed by evaporation or lyophilization.

[0190] NMR method

[0191] NMR spectra were obtained using standard Bruker pulse sequences on a Bruker Avance III 600 (5 mm RT inverting probe), Bruker DRX 500, Bruker Avance AV 400 (5 mm RT directional probe), or Bruker DPX 300 spectrometer. Alternatively, NMR spectra were recorded using a Varian MR-400 spectrometer operating at 400 MHz or a Varian Unity Inova 400 spectrometer with a 5 mm inverting detection triple resonance probe operating at 400 MHz. DMSO - d 6 or CDCl 3 is used as the solvent, and tetramethylsilane is used as the internal standard, except in the latter case where the solvent residue peak is used. All experiments are recorded at 298 K unless otherwise described. Chemical shifts are given relative to the internal standard tetramethylsilane or the solvent residue peak. Coupling constants (J values) are given in Hertz (Hz), and multiplicity is reported using the following abbreviations: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broad peak, nd = not determined.

[0192] LCMS Method 1

[0193] Acquity UPLC coupled to SQD mass spectrometer; column: Acquity UPLC BEH C 18 (50mm x 2.1mm i.d., 1.7μm packing diameter), mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile.

[0194] gradient

[0195]

[0196] Column temperature: 40℃; UV detection: from 210 nm to 350 nm; MS conditions: ionization mode: alternating scan electrospray ionization (ESI) + / ES - ), Scan range: 100 to 1000 AMU.

[0197] LCMS Method 2

[0198] Acquity UPLC coupled to SQD mass spectrometer; column: Acquity UPLC BEH C 18 (50 mm x 2.1 mm i.d., 1.7 μm packing diameter), mobile phase A: 10 mM ammonium bicarbonate aqueous solution (adjusted to pH 10 with ammonia), mobile phase B: acetonitrile;

[0199] gradient

[0200]

[0201] Column temperature: 40℃; UV detection: from 210 nm to 350 nm; MS conditions: ionization mode: alternating scan electrospray ionization (ESI) + / ES - ), Scan range: 100 to 1000 AMU.

[0202] LCMS Method 3

[0203] AGILENT LC 1260 Infinity with SFC and Agilent 6540 UHD Accurate-Mass Q-TOF LC / MS; Column: Acquity UPLC BEH C 18 (100mm x 2.1mm id, 1.7μm packing diameter), mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile.

[0204] gradient

[0205]

[0206] Column temperature: 40℃; UV detection: from 210 nm to 350 nm; MS conditions: ionization mode: alternating scan electrospray ionization (ESI) + / ES - ), Scan range: 100 to 1500 AMU.

[0207] Abbreviations used:

[0208] Boc2O = di-tert-butyl dicarbonate; aq. = water (solution); DCC = dicyclohexylcarbodiimide; DCM = dichloromethane; DIPEA = N,N -Diisopropylethylamine; DMAP = 4-Dimethylaminopyridine; DMCHDA = trans - N,N′ -Dimethylcyclohexane-1,2-diamine; DMF = N,N -Dimethylformamide; DMSO = dimethyl sulfoxide; EtOAc = ethyl acetate; LCMS = liquid chromatography-mass spectrometry; 1 H-NMR = proton nuclear magnetic resonance; RM = reaction mixture; Rt = residence time; RT = room temperature; sat. = saturation; TEA = triethylamine; TFA = trifluoroacetic acid; THF = tetrahydrofuran; Xphos-Pd-G3-(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate.

[0209] In subsequent processes, some starting materials are identified by "intermediate" or "example" designations and noted in the step number. This is provided merely to assist skilled chemists.

[0210] "Similar" or "resembling" procedures mean that such procedures may involve minor variations, such as reaction temperature, reagent / solvent volume, reaction time, post-treatment conditions, or chromatographic purification conditions.

[0211] The stereochemistry of the compounds in the examples (where specified) has been specified to assume that the absolute configuration at the center of the split stereoisomer source of the starting material remains unchanged under any subsequent reaction conditions.

[0212] Unless otherwise stated, when the absolute configuration (R) or (S) is reported in the compound name, ee% must be considered equal to or greater than 90%.

[0213] Preparation of intermediates

[0214] Intermediate 1

[0215] Step 1

[0216]

[0217] 7-Bromo-6-methoxy-3,4-dihydro-2 H- Benzo[ b [1,4]Oxazine (intermediate 1-1)

[0218] 6-methoxy-3,4-dihydro- 2H A solution of 1,4-benzoxazine (3.0 g, 18.20 mmol) in EtOAc (30.0 mL) was cooled to 0 °C. 1,3-Dibromo-5,5-dimethylimidazolidine-2,4-dione (2.6 g, 9.08 mmol) was added in portions over 15 minutes. RM was stirred at 0 °C for an additional 30 min and quenched with an aqueous solution of K₂CO₃ (10% w / w; 60 mL). The organic layer was separated, washed with sat. aq. NaCl, and concentrated under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0–30% EtOAc in cyclohexane to provide the title product (3.5 g).

[0219] LCMS (Method 1): Rt = 0.97 min, ES + m / z 243.9 / 245.9 [M+H] + .

[0220] Step 2

[0221]

[0222] 7-Bromo-6-methoxy-2,3-dihydro-4 H- Benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 1)

[0223] THF (15 mL) was added to a mixture of intermediate 1-1 (1.4 g, 5.74 mmol), DMAP (840.9 mg, 6.88 mmol), and Boc2O (2.80 g, 13.19 mmol), and RM was stirred overnight at RT. RM was then partitioned between EtOAc (50 mL) and water (30 mL). The organic layer was washed with 2 M aq. citric acid (2 x 20 mL), sat. aq. NaCl (20 mL), and evaporated under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0–10% EtOAc in cyclohexane to provide the title compound (1.26 g).

[0224] LCMS (Method 1): Rt = 1.27 min

[0225] 1 H-NMR (300 MHz, CDCl 3) δ: 7.56 (brs, 1H), 7.05 (s, 1H); 4.14-4.18 (m, 2H), 3.82 (s, 3H), 3.78-3-82 (m, 2H), 1.53 (s, 9H).

[0226] Intermediate 2

[0227] Step 1

[0228]

[0229] 6-Methoxy-2,3-dihydro-4H-benzo[b][1,4]oxazine-4-carboxylic acid tert-butyl ester (intermediate 2-1)

[0230] A solution of 7-methoxy-1,2,3,4-tetrahydroquinoline (500 mg, 3.06 mmol), DMAP (449 mg, 3.68 mmol), and Boc₂O (1.54 g, 7.05 mmol) in THF (10 mL) was stirred overnight at RT. Another equivalent of Boc₂O was added and stirring continued at RT. RM was partitioned between EtOAc (50 mL) and water (30 mL). The organic layer was washed with aq. 2 M citric acid (2 x 15 mL), sat. aq. NaCl (20 mL), and the solvent was evaporated under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0–10% EtOAc in cyclohexane to provide the title product (228 mg).

[0231] LCMS (Method 5): Rt = 2.5 min

[0232] 1 H-NMR (300 MHz, DMSO - d 6 ) δ: 7.47 - 7.38 (m, 1H), 6.77 (d, J = 8.9 Hz,1H), 6.57 (dd, J = 8.9, 3.0 Hz, 1H), 4.18 - 4.09 (m, 2H), 3.80 - 3.72 (m,2H), 3.68 (s, 3H), 1.50 (s, 9H).

[0233] Step 2

[0234]

[0235] 7-Iodo-6-methoxy-2,3-dihydro-4H-benzo[b][1,4]oxazine-4-carboxylic acid tert-butyl ester (intermediate 2)

[0236] Intermediate 2-1 (5.9 g, 22.1 mmol) was dissolved in DMF (60 mL), then N-iodohydroxysuccinimide (12.7 g, 111 mmol) was added and RM was stirred overnight at 40 °C. RM was quenched in cold water and extracted with EtOAc. The combined organic layers were evaporated to dryness and the residue was purified by chromatography on silica gel by a gradient elution of 1:1 to 3:2 DCM-hexane to provide the title compound (7.38 g).

[0237] LCMS (Method 1): Rt = 2.8 min

[0238] 1 H-NMR (300 MHz, DMSO - d 6 ) δ: 7.50 (s, 1H), 7.24 (s, 1H), 4.14 (dd, J =5.3, 3.8 Hz, 2H), 3.80 - 3.75 (m, 2H), 3.74 (s, 3H), 1.51 (s, 9H).

[0239] Intermediate 3

[0240] Step 1

[0241]

[0242] 1-Bromo-5-fluoro-2-methoxy-4-nitrobenzene (Intermediate 3-1)

[0243] A mixture of 2-bromo-4-fluoro-5-nitrophenol (2.5 g, 11 mmol), K₂CO₃ (2.20 g, 16 mmol), and iodomethane (923 µL, 15 mmol) in DMF (9.3 mL) was stirred at RT for 1.5 h. RM was poured into water (40–50 mL), and the precipitate was filtered, washed with water, and dried to provide the title product (2.58 g).

[0244] LCMS (Method 1): Rt = 1.13 min

[0245] 1 H-NMR (500 MHz, DMSO-d 6) δ: 8.06 (d, J=10.5 Hz, 1H), 7.76 (d, J=6.5Hz, 1H), 3.95 (s, 3H).

[0246] Step 2

[0247]

[0248] 2-((5-bromo-4-methoxy-2-nitrophenyl)thio)methyl acetate (intermediate 3-2)

[0249] A solution of intermediate 3-1 (2.1 g, 8.2 mmol) and DIPEA (1.43 mL, 8.2 mmol) in acetonitrile (40 mL) cooled to 0 °C was added dropwise over 1 h to methyl thioglycolate (736 µL, 8.2 mmol) in acetonitrile (28 mL). RM was stirred at 0 °C for 2 h, and then the solvent was evaporated under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0–100% EtOAc / cyclohexane (3:7)cyclohexane to provide the title product (2.31 g).

[0250] LCMS (Method 1): Rt = 1.13 min, ES + m / z 336.1 / 338.1 [M+H] + .

[0251] Step 3

[0252]

[0253] 7-Bromo-6-methoxy-2H-benzo[ b [1,4]Thiazine-3(4 H )-Ketone (Intermediate 3)

[0254] Aq. NH4Cl (734 mg, 14 mmol, 15 mL) and iron powder (2.3 g, 41 mmol) were added to a warm solution of intermediate 3-2 (1.4 g, 3.4 mmol) in ethanol (28 mL) at 80 °C, and the mixture was stirred at 85 °C for 36 h. After cooling to RT, the mixture was diluted with water, filtered to remove undissolved solids, and the filtrate was extracted with EtOAc (3x). The combined organic matter was washed with aq. NaCl (2x) and evaporated under vacuum. The residue was ground with methanol to provide the title product (574 mg).

[0255] LCMS (Method 1): Rt = 0.98 min, ES + m / z 273.9 / 276.0 [M+H] + .

[0256] Intermediate 4a

[0257]

[0258] 5-((dimethylamino)methyl)oxazolidin-2-one (intermediate 4a)

[0259] A vial containing 1.0 g (7.4 mmol) of 5-(chloromethyl)oxazolidin-2-one and 13.0 mL (26.0 mmol) of dimethylamine (2.0 M in methanol, 13.0 mL) was heated at 150 °C under microwave irradiation for 30 min. After cooling to RT, RM was concentrated under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0–100% DCM:MeOH:NH4OH (90:15:1.5) in DCM. The title product (399 mg) was obtained in solid form.

[0260] 1 H-NMR (500 MHz, DMSO-d 6 ) δ: 7.44 (brs, 1H), 4.61-4.67 (m, 1H), 3.50 (t, J=8.6 Hz, 1H), 3.13 (t, J=9.0 Hz, 1H), 2.94 (dd, J=13.0, 6.5 Hz, 1H), 2.43 (dd, J=13.0, 5.5 Hz, 1H), 2.18 (s, 6H).

[0261] Intermediate 4b

[0262]

[0263] ( S )-5-((dimethylamino)methyl)oxazolidin-2-one (intermediate 4b)

[0264] The title compound was prepared starting from (S) 5-(chloromethyl)oxazolidin-2-one in a manner similar to that of intermediate 4a.

[0265] 1 H-NMR (300 MHz, DMSO-d 6 ) δ: 6.75 (bs, 1H); 4.70-4.58 (m, 1H); 3.58 (t,J=8.9 Hz, 1H); 3.23 (t, J=7.9 Hz, 1H); 2.60-2.40 (m, 2H); 2.22 (s, 6H).

[0266] Intermediate 5

[0267] Step 1

[0268]

[0269] 7-(3-amino-6-chloro-1) H- Pyrazolo[4,3] -c ]pyridin-1-yl)-6-methoxy-2,3-dihydro-4 H- benzo[a] [ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 5-1)

[0270] Intermediate 1 (408 mg, 1.19 mmol), 6-chloro- 1H -pyrazolo[4,3- c A degassed mixture of pyridine-3-amine (200 mg, 1.19 mmol), Cs₂CO₃ (980 mg, 3.01 mmol), DMCHDA (93.5 µL, 0.59 mmol), and CuI (113 mg, 0.59 mmol) in DMSO (4.9 mL) was stirred overnight at 110 °C under argon. After cooling to RT, RM was quenched with sat. aq. NaHCO₃ (50 mL) and extracted with EtOAc (5 x 50 mL). The combined organic layers were washed with sat. aq. NaCl, dried over Na₂SO₄, and the solvent was removed under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0–50% DCM / MeOH / NH₄OH (90:5:0.5) in DCM to provide the title product (282 mg).

[0271] LCMS (Method 1): Rt = 1.15 min, ES + m / z 431.9 / 433.9 [M+H] + .

[0272] Step 2

[0273]

[0274] 7-(6-chloro-3-(((2-chloroethoxy)carbonyl)amino)-1 H- Pyrazolo[4,3] -c] pyridin-1-yl)-6-methoxy 2,3-dihydro-4-yl H- Benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 5-2)

[0275] Intermediate 5-1 (66 mg, 0.15 mmol) and pyridine (16 µL, 0.20 mmol) in a pre-cooled (0 °C) solution of DCM (0.96 mL) were supplemented with 2-chloroethyl chloroformate (17 µL, 0.17 mmol), and RM was stirred in an ice bath for 30 min and at RT for 1 h. RM was partitioned between DCM (20 mL) and sat aq. NaHCO3, and the aqueous layer was further extracted with DCM (3 × 20 mL). The combined organic layers were washed with sat. aq. NaHCO3 and sat. aq. NaCl, dried over Na2SO4, and the solvent was removed under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0–100% DCM / MeOH / NH4OH (90:5:0.5) in DCM. The isolated material was stirred overnight in DCM / MeOH / NH4OH (90:5:0.5), and then the solvent was evaporated under vacuum to provide the title product (94 mg), which was used for subsequent synthetic steps without further purification.

[0276] LCMS (Method 1): Rt = 1.34 min, ES + m / z 538.1 / 540.1 / 542.2 [M+H] + .

[0277] Step 3

[0278]

[0279] 7-(6-chloro-3-(2-oxomylidene-3-yl)-1 H- Pyrazolo[4,3] -c] pyridin-1-yl)-6-methoxy- 2,3-Dihydro-4 H- Benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 5)

[0280] A solution of intermediate 5-2 (94.0 mg, 0.13 mmol) and morpholine (11.7 µL, 0.14 mmol) in DMF (1.3 mL) was stirred at RT for 1 h, then at 80 °C for 1 h. After cooling to RT, the RM was diluted with sat. aq. NaHCO3 and extracted with EtOAc (3 × 20 mL). The combined organics were washed with sat. aq. NaCl, dried over Na2SO4, and the solvent was removed under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0–50% EtOAc in cyclohexane to provide the title product (40 mg).

[0281] LCMS (Method 2): Rt = 1.28 min, ES + m / z 502.2 / 504.2 [M+H] + .

[0282] Intermediate 6a

[0283] Step 1

[0284]

[0285] 6-Chloro-3-iodo-1-(tetrahydro-2-) H- pyran-2-yl)-1 H- Pyrazolo[4,3] -c] Pyridine (intermediate 6a-1)

[0286] Dihydropyran (9.79 mL, 107 mmol) and methanesulfonic acid (464 μL, 7.16 mmol) were added to 6-chloro-3-iodine-1-dihydropyran in DCM (100 mL) and THF (50 mL). H -Pyrazolo[4,3-c]pyridine (10 g, 35.8 mmol). RM was stirred at 40 °C for 4 h, then stirred overnight at RT. RM was evaporated to dryness and the residue was purified by rapid chromatography on a Si column by elution with 0-40% EtOAc in cyclohexane to provide the title product (7.5 g).

[0287] LCMS (Method 2): Rt = 1.22 min, ES + m / z 364.0 / 366.0 [M+H] + .

[0288] Step 2

[0289]

[0290] 3-(6-chloro-1-(tetrahydro-2-) H- pyran-2-yl)-1 H- Pyrazolo[4,3] -c] pyridin-3-yl)-5-((dimethylamino) (Methyl)oxazolidin-2-one (intermediate 6a-2)

[0291] A degassed mixture of intermediate 6a-1 (1.0 g, 2.75 mmol), intermediate 4a (793.0 mg, 5.50 mmol), K₂CO₃ (2.28 g, 16.5 mmol), and CuI (210.0 mg, 1.10 mmol) in DMSO (10.0 mL) was stirred at 90 °C for 16 h. The solvent was removed under vacuum at 60 °C, and the residue was partitioned between EtOAc (10 mL) and water (10 mL). The aqueous phase was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with water (2 x 10 mL) and sat. aq. NaCl (10 mL), dried over MgSO₄, and the solvent was removed under vacuum. The residue was purified by rapid chromatography on a Si column by elution with DCM / MeOH (20:1) to provide the title product (857 mg).

[0292] LCMS (Method 1): Rt = 0.65 min, ES + m / z 380.2 / 382.2 [M+H] + .

[0293] Step 3

[0294]

[0295] 3-(6-chloro-1) H- Pyrazolo[4,3] -c] Pyridin-3-yl)-5-((dimethylamino)methyl)oxazolidin-2-one (middle) Interstitial 6a-3)

[0296] A solution of intermediate 6a-2 (855 mg, 2.14 mmol) in DCM (10 mL) was treated with TFA (2.52 mL, 32.9 mmol) and Et3SiH (1.03 mL, 6.42 mmol) at RT for 16 h. A second portion of TFA (1.26 mL, 16.45 mmol) and Et3SiH (341.7 μL, 2.14 mmol) was added, and RM was stirred at RT for another 16 h. RM was pre-purified on an SCX column to provide crude material, which was then subjected to rapid chromatography on a Si column using 0–80% DCM / MeOH / NH4OH (90:9:1.5) to provide the title product (430 mg).

[0297] LCMS (Method 2): Rt = 0.62 min, ES + m / z 296.1 / 298.1 [M+H] + .

[0298] Step 4

[0299]

[0300] 7-(6-chloro-3-(5-((dimethylamino)methyl)-2-oxomylidene-3-yl)-1 H- Pyrazolo[4,3] -c] pyridin-1-yl)-6-methoxy-2,3-dihydro-4 H- Benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 6a)

[0301] Intermediate 6a-3 (260 mg, 0.84 mmol), intermediate 2 (330.0 mg, 0.84 mmol), K2CO3 (350.0 mg, 2.53 mmol), N,NA degassed solution of dimethylglycine (131.0 mg, 1.27 mmol) and CuI (80.4 mg, 0.42 mmol) in DMSO (8 mL) was stirred at 110 °C for 24 h. The solvent was removed under vacuum at 40 °C. The residue was purified by rapid chromatography on a Si column by elution with 0–100% DCM / MeOH (20:1) in DCM to provide the title product (294.5 mg).

[0302] LCMS (Method 2): Rt = 1.30 min, ES + m / z 559.2 / 561.2 [M+H] + .

[0303] Intermediate 6b

[0304] Step 1

[0305]

[0306] (5 R )-3-(6-chloro-1-(tetrahydro-2-) H- pyran-2-yl)-1 H- Pyrazolo[4,3] -c] pyridin-3-yl)-5-((di) (Methylamino)methyl)oxazolidin-2-one (intermediate 6b-1)

[0307] The title product is prepared starting from intermediates 6a-1 and 4b in a manner similar to that of intermediate 6a-2.

[0308] LCMS (Method 2): Rt = 1.00 min, ES + m / z 380.2 / 382.2 [M+H] + .

[0309] Step 2

[0310]

[0311] ( R )-3-(6-chloro-1 H- Pyrazolo[4,3] -c] pyridin-3-yl)-5-((dimethylamino)methyl)oxazolidine-2- Ketone (intermediate 6b-2)

[0312] The title product is prepared starting from intermediate 6b-1 in a manner similar to that of intermediate 6a-3.

[0313] LCMS (Method 2): Rt = 0.56 min, ES + m / z 296.1 / 298.1 [M+H] + .

[0314] Step 3

[0315]

[0316] ( R )-7-(6-chloro-3-(5-((dimethylamino)methyl)-2-oxomylidene-3-yl)-1 H- Pyrazolo[4, 3 -c] pyridin-1-yl)-6-methoxy-2,3-dihydro-4 H- Benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 6b)

[0317] The title product is prepared starting from intermediates 6b-2 and 2 in a manner similar to step 4 of intermediate 6a.

[0318] LCMS (Method 2): Rt = 1.30 min, ES + m / z 559.2 / 561.2 [M+H] + .

[0319] Intermediate 7

[0320]

[0321] 7-(6-chloro-3-(5-(((methylamino)methyl)-2-oxomylidene-3-yl)-1 H- Pyrazolo[4,3] -c] pyrene (Pyridine-1-yl)-6-methoxy-2,3-dihydro-4 H- Benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 7)

[0322] The intermediate 6a (30 mg, 0.05 mmol), sodium acetate (12.1 mg, 0.15 mmol), and iodine (20.0 mg, 0.08 mmol) in MeOH (1 mL) were stirred and irradiated with a mercury lamp (28 W) for 16 h. RM was partitioned between sat.aq. Na2S2O3 (10 mL) and DCM (5 mL), and the aqueous layer was further extracted with DCM (5 x 10 mL). The combined organic layers were passed through a phase separator column and evaporated to dryness. The residue was purified by rapid chromatography on a Si column by elution with 0-100% DCM / MeOH:NH4OH (90:9:1.5) in DCM to provide the title product (17 mg).

[0323] LCMS (Method 2): Rt = 1.24 min, ES + m / z 545.2 / 547.2 [M+H] + .

[0324] Intermediate 8a

[0325] Step 1

[0326]

[0327] 1-(6-chloro-1-(tetrahydro-2-) H -pyran-2-yl)-1 H -pyrazolo[4,3- c ]pyridin-3-yl)-3-methylimidazolium Alkyl-2-one (intermediate 8a-1)

[0328] A degassed mixture of intermediate 6a-1 (332 mg, 0.91 mmol), 1-methylimidazolidine-2-one (274 mg, 2.74 mmol), 3,4,7,8-tetramethyl-1,10-phenanthroline (54 mg, 0.23 mmol), CuI (35 mg, 0.18 mmol), and K3PO4 (581 mg, 2.73 mmol) in DMSO (3.5 mL) was stirred at 110 °C for 2.5 h. Another RM was prepared in a similar manner, starting with 270 mg of intermediate 6a-1. After cooling to RT, the combined reaction mixture was diluted with water and extracted with DCM. The organic layer was washed with sat. aq. NaCl, dried over Na2SO4, and the solvent was removed under vacuum. Residual DMSO was removed on a GeneVac. The residue was purified by rapid chromatography on a Si column by elution with 0–40% EtOAc in DCM to provide the title product (420 mg).

[0329] LCMS (Method 2): Rt = 1.01 min, ES + m / z 336.2 / 338.2 [M+H] + .

[0330] Step 2

[0331]

[0332] 1-(6-chloro-1H-pyrazolo[4,3-c]pyridin-3-yl)-3-methylimidazolidine-2-one (intermediate 8a-2)

[0333] The title product is prepared starting from intermediate 8a-1 in a manner similar to that of intermediate 6a-3.

[0334] LCMS (Method 2): Rt = 0.63 min, ES + m / z 252.1 / 254.1 [M+H] + .

[0335] Step 3

[0336]

[0337] 7-(6-chloro-3-(3-methyl-2-oxomylidene-1-yl)-1 H -pyrazolo[4,3-c]pyridin-1-yl)-6- Methoxy-2,3-dihydro-4 H -benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 8a)

[0338] The title product is prepared starting from intermediates 8a-2 and 2 in a manner similar to step 4 of intermediate 6a-.

[0339] LCMS (Method 2): Rt = 1.30 min, ES+ m / z 515.2 / 517.2 [M+H] + .

[0340] Intermediate 8b

[0341]

[0342] 7-(6-chloro-3-(3-methyl-2-oxomylidene-1-yl)-1 H -pyrazolo[4,3-c]pyridin-1-yl)-6- Methoxy-2 H -benzo[ b [1,4]Thiazine-3(4 H )-Ketone (Intermediate 8b)

[0343] Starting with intermediates 8a-2 and 3, to... Step 4 of intermediate 6a The title product was prepared in a similar manner.

[0344] LCMS (Method 2): Rt = 0.90 min, ES + m / z 445.1 / 447.0 [M+H] +

[0345] intermediate 8c

[0346]

[0347] 1-(6-chloro-1-(6-methoxy-3,4-dihydro-2-) H -benzo[ b [1,4]thiazine-7-yl)-1 H -pyrazolo[4,3- c ]pyridin-3-yl)-3-methylimidazolidine-2-one (intermediate 8c)

[0348] BH3·THF (1.00 M, 230 µL, 0.23 mmol in THF) was added dropwise to an ice-bath-cooled suspension of intermediate 8b (46.0 mg, 92 μmol, 89%) in dry THF (1.4 mL) under Ar conditions. RM was stirred at RT for 2 h. RM was cooled again in an ice bath and quenched with sat. aq. NaHCO3. RM was extracted with DCM (2 x). The combined organic layers were washed with water, sat. aq. NaCl, dried over Na2SO4, and the solvent was removed under vacuum. The residue was purified twice by rapid chromatography on a Si column, first by elution with an increased amount of MeOH in DCM and second by elution with DCM / EtOAc (1:1) to provide the title product (27 mg).

[0349] LCMS (Method 1): Rt = 1.04 min, ES + m / z 431.3 / 433.0 [M+H] +

[0350] Intermediate 9

[0351] Step 1

[0352]

[0353] 1-(4-(tert-Butoxycarbonyl)-6-methoxy-3,4-dihydro-2 H- Benzo[ b [1,4]oxazine-7-yl)-6-chloro- 1 H- Pyrazolo[4,3] -c] Pyridine-3-carboxylic acid (intermediate 9-1)

[0354] A degassed mixture of 6-chloro-1H-pyrazolo[4,3-c]pyridine-3-carboxylic acid (9.4 g, 47.5 mmol), intermediate 2 (18.6 g, 47.5 mmol), Cs₂CO₃ (54 g, 166.3 mmol), and thiophene-2-carbonyloxycopper (9 g, 47.5 mmol) in DMSO (100 mL) was stirred at 110 °C for 24 h under argon. RM was quenched in water and extracted with DCM. The combined organic layers were washed with aq. 10% w / w citric acid and aq. sat. NaCl and evaporated to dryness. The residue was purified by chromatography on silica gel by elution with DCM to DCM (1% v / v MeOH + 1% AcOH v / v). The resulting substance was ground in diethyl ether to provide the title compound (3.14 g).

[0355] LCMS (Method 1): Rt = 1.22 min, ES + m / z 461.2 / 463.2 [M+H] + .

[0356] Step 2

[0357]

[0358] 7-(3-(((benzyloxy)carbonyl)amino)-6-chloro-1 H- Pyrazolo[4,3] -c] pyridin-1-yl)-6-methoxy-2, 3-Dihydro-4 H- Benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 9-2)

[0359] Intermediate 9-1 (1.0 g, 2.2 mmol), propanephosphonic anhydride (50% solution in EtOAc, 2.53 mL, 4.3 mmol), TMS-N3 (576 µL, 4.3 mmol), and TEA (907 µL, 6.5 mmol) were refluxed in 2-Me THF (40 mL) for 1 h. Benzyl alcohol (1.12 mL, 11 mmol) was added, and RM was refluxed for another 48 h. After cooling to RT, RM was diluted with EtOAc and sat. aq. NaHCO3. The aqueous layer was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with sat. aq. NaCl, dried over Na2SO4, and concentrated under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0–25% EtOAc in cyclohexane to provide the title product (952 mg).

[0360] LCMS (Method 1): Rt = 1.39 min, ES + m / z 566.2 / 568.2 [M+H] + .

[0361] Step 3

[0362]

[0363] 7-(6-chloro-3-(5-(2-(1,3-dioxylideneisoindoline-2-yl)ethyl)-2-oxylideneoxazolidine-3- (base)-1 H- Pyrazolo[4,3] -c] pyridin-1-yl)-6-methoxy-2,3-dihydro-4 H- Benzo[ b [1,4]oxazine-4-carboxylic acid tert-oxazine Butyl ester (intermediate 9-3)

[0364] LiHMDS (1.3 M in THF, 576.0 µL, 0.75 mmol) was added to a 0°C cooled solution of intermediate 9-2 (212.0 mg, 0.375 mmol) in THF (4 mL) and the solution was stirred at 0°C for 1 h. 2-[2-(ethylene oxide-2-yl)ethyl]isoindoline-1,3-dione (163.0 mg, 0.75 mmol) was added and the solution was stirred at 110°C for 16 h. After cooling to RT, the solution was carefully diluted with cold water, DCM, and sat. aq. NaHCO3. The layers were separated, and the aqueous layer was diluted with DCM / i- PrOH (1:1, 4 × 15 mL) was extracted separately. The combined organic layers were passed through a phase separator column and the solvent was removed under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0-100% EtOAc in DCM to provide the title product (159 mg).

[0365] LCMS (Method 2): Rt = 1.40 min, ES + m / z 675.3 / 677.3 [M+H] + .

[0366] Step 4

[0367]

[0368] 7-(3-(5-(2-aminoethyl)-2-oxomylidene-3-yl)-6-chloro-1 H- Pyrazolo[4,3] -c] Pyridine- 1-yl)-6-methoxy-2,3-dihydro-4 H- Benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 9-4)

[0369] Intermediate 9-3 (133 mg, 0.20 mmol) was treated with hydrazine hydrate (248 µL, 5.12 mmol) in EtOH (2.0 mL) and stirred at 25 °C for 2 h. The precipitate was filtered and the filtrate was partitioned between EtOAc (30 mL) and water (10 mL). The organic layer was washed with sat. aq. NaCl (10 mL) and the solvent was removed under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0-100% DCM / MeOH / NH4OH (90:9:1.5) in DCM to provide the title product (44 mg).

[0370] LCMS (Method 1): Rt = 1.22 min, ES + m / z 545.2 / 547.2 [M+H] + .

[0371] Step 5

[0372]

[0373] 7-(6-chloro-3-(5-(2-(dimethylamino)ethyl)-2-oxomylidene-3-yl)-1 H- Pyrazolo[4,3] - c] pyridin-1-yl)-6-methoxy-2,3-dihydro-4 H- Benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 9)

[0374] At RT, NaB(CN)H3 (16.1 mg, 0.26 mmol) was added to a mixture of intermediate 9-4 (35.0 mg, 0.06 mmol), 4 Å molecular sieve (powder, 50 mg), formaldehyde (37% in water, 10 μL, 0.14 mmol), and acetic acid (5.5 μL, 0.10 mmol) in methanol (1 mL), and RT was stirred for 1 h. RT was quenched with water (5 mL) and washed with EtOAc (2 x 10 mL). The pH of the aqueous layer was adjusted to 11 with aq. 1.0 M NaOH and extracted with EtOAc (3 x 10 mL). The combined organic layers from the last extraction were passed through a phase separator column and the solvent was removed under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0-100% DCM / MeOH / NH4OH (90:9:0.5) in DCM to provide the title product (33 mg).

[0375] LCMS (Method 2): Rt = 1.33 min, ES + m / z 573.3 / 575.3 [M+H] + .

[0376] Intermediate 10

[0377] Step 1

[0378]

[0379] ( S )-7-(6-chloro-3-(5-(((1,3-dioxylideneisoindoline-2-yl)methyl)-2-oxylideneoxazolidine-3- (base)-1 H -pyrazolo[4,3- c ]pyridin-1-yl)-6-methoxy-2,3-dihydro-4 H -benzo[ b [1,4]oxazine-4-carboxylic acid tert-oxazine Butyl ester (intermediate 10-1)

[0380] Starting from intermediates 9-2 and 2-[[(2S)-epoxyethylene-2-yl]methyl]isoindoline-1,3-dione, to... middle Interstitial 9-3 The title product was prepared in a similar manner.

[0381] LCMS (Method 1): Rt = 1.35 min, ES + m / z 661.2 / 663.2 [M+H] + .

[0382] Step 2

[0383]

[0384] ( S )-7-(3-(5-(aminomethyl)-2-oxadiazolidin-3-yl)-6-chloro-1 H -pyrazolo[4,3- c ]pyrene (Pyridine-1-yl)-6-methoxy-2,3-dihydro-4 H -benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 10-2)

[0385] Starting with intermediate 10-1, to... Intermediate 9-4 The title product was prepared in a similar manner.

[0386] LCMS (Method 1): Rt = 0.94 min, ES + m / z 531.2 / 533.2 [M+H] + .

[0387] Step 3

[0388]

[0389] ( S )-7-(6-chloro-3-(5-((((2-methoxyethyl)amino)methyl)-2-oxomylidene-3-yl)-1 H - Pyrazolo[4,3- c ]pyridin-1-yl)-6-methoxy-2,3-dihydro-4 H -Benzo[b][1,4]oxazine-4-tert-butyl carboxylate (middle) Interstitial 10-3)

[0390] A solution of intermediate 10⁻² (66 mg, 0.12 mmol), 1-bromo-2-methoxy-ethane (17 mg, 0.12 mmol), and DIPEA (86 µL, 0.49 mmol) in DMF (1.6 mL) was stirred overnight at 60 °C. Another equivalent of 1-bromo-2-methoxy-ethane was added, and the solution was stirred at 60 °C for another 48 h. After cooling to RT, the solution was partitioned between EtOAc and water. The organic layer was washed with sat. aq. NaCl and dried over Na₂SO₄. The organic layer was evaporated under vacuum, and the residue was purified by rapid chromatography on a Si column by elution with 0–50% DCM / MeOH / NH₄OH (90:9:0.5) in DCM to provide the title product (45.5 mg).

[0391] LCMS (Method 1): Rt = 1.22 min, ES + m / z 589.2 / 591.2 [M+H] + .

[0392] Step 4

[0393]

[0394] ( S )-7-(6-chloro-3-(5-(((2-methoxyethyl)(methyl)amino)methyl)-2-oxadiazolidine-3- (base)-1 H -pyrazolo[4,3-c]pyridin-1-yl)-6-methoxy-2,3-dihydro-4 H -benzo[ b [1,4]oxazine-4-carboxylic acid tert-oxazine Butyl ester (intermediate 10)

[0395] NaCNBH3 (19.2 mg, 0.31 mmol) was added to a mixture of intermediate 10⁻³ (45.0 mg, 76.4 μmol), formaldehyde (37.0% in water, 12.4 µL, 0.17 mmol), acetic acid (6.56 µL, 0.12 mmol), and molecular sieve (4 Å, dry powder, 50 mg) in MeOH (1.2 mL), and the mixture was stirred at 25 °C for 1 h. The reaction was quenched with water (10 mL) and extracted with EtOAc (3 x 15 mL). The combined organic layers were dried over Na₂SO₄ and the solvent was removed under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0–40% DCM / MeOH / NH₄OH (90:9:0.5) in DCM to provide the title product (29 mg).

[0396] LCMS (Method 1): Rt = 0.99 min, ES + m / z 603.3 / 605.2 [M+H] + .

[0397] Intermediate 11

[0398] Step 1

[0399]

[0400] ( S )-7-(6-chloro-3-((2,3-dihydroxypropyl)amino)-1 H- Pyrazolo[4,3] -c] pyridin-1-yl)-6-methyl oxy-2,3-dihydro-4 H- Benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 11-1)

[0401] The solution of intermediate 9-2 (85.0 mg, 0.15 mmol) in THF (1.7 mL) was cooled to 0 °C and treated with NaH (60.0% suspension in mineral oil, 18.0 mg, 0.45 mmol). RM was stirred at RT for 60 min, then cooled to 0 °C, followed by the addition of butyric acid [(2 S [(2S)-ethylene oxide-2-yl]methyl butyrate (42.9 µL, 0.30 mmol). RM was stirred at RT for 48 h. A second equivalent of [(2S)-ethylene oxide-2-yl]methyl butyrate (42.9 µL, 0.30 mmol) was added and RM was stirred for another 24 h. RM was quenched with cold water and partitioned between DCM and NaHCO3. The aqueous layer was further extracted with DCM (3 × 15 mL). The combined organic layers were dried over Na2SO4 and the solvent was removed under vacuum. The residue was redissolved in MeOH, a small amount of K2CO3 was added, and the mixture was stirred at RT overnight. The crude product was purified by rapid chromatography on a Si column by elution with 0–100% DCM / MeOH (9:1) in DCM to provide the title product (22 mg).

[0402] LCMS (Method 2): Rt = 1.05 min, ES + m / z 506.0 / 507.9 [M+H] + .

[0403] Step 2

[0404]

[0405] ( S )-7-(6-chloro-3-(5-(hydroxymethyl)-2-oxomylidene-3-yl)-1 H- Pyrazolo[4,3] -c] pyrene (Pyridine-1-yl)-6-methoxy-2,3-dihydro-4 H- Benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 11)

[0406] Intermediate 11-1 (25.0 mg, 49 µmol), KO tA mixture of Bu (5.5 mg, 49 µmol) and diethyl carbonate (12 µL, 99 µmol) in toluene (1.7 mL) was refluxed overnight. The solvent was removed under vacuum and the residue was eluted on a Si column by rapid chromatography with 0–80% DCM / MeOH (10:0.4) in DCM to provide the title product (10 mg).

[0407] LCMS (Method 2): Rt = 1.16 min, ES + m / z 532.0 / 533.9 [M+H] + .

[0408] Intermediate 12

[0409] Step 1

[0410]

[0411] 7-(6-chloro-3-(3-(2-chloroethyl)ureido)-1 H -pyrazolo[4,3- c ]pyridin-1-yl)-6-methoxy-2,3- Dihydro-4 H -benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 12-1)

[0412] Under argon atmosphere, a mixture of intermediate 5-1 (200 mg, 0.46 mmol), 1-chloro-2-isocyanate-ethane (158 µL, 1.85 mmol), and DIPEA (323 µL, 1.85 mmol) in dry DCM (5 mL) was stirred overnight at RT. RM was quenched with sat. aq. NaHCO3 and extracted with DCM. The combined organic layers were washed with water, sat. aq. NaCl, dried over Na2SO4, and the solvent was evaporated under vacuum to provide the title product (336 mg), which was used for subsequent synthetic steps without further purification.

[0413] LCMS (Method 1): Rt = 1.26 min, ES + m / z 537.1 / 539.1 / 541.1 [M+H] + .

[0414] Step 2

[0415]

[0416] 7-(6-chloro-3-(2-oxomylidene-1-yl)-1 H -pyrazolo[4,3- c ]pyridin-1-yl)-6-methoxy- 2,3-Dihydro-4 H -benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 12-2)

[0417] Intermediate 12-1 (326 mg, 0.51 mmol) and KO tThe mixture of Bu (68.3 mg, 0.61 mmol) in dry THF (5 mL) was stirred overnight at RT under argon. The reaction was quenched with water and extracted with DCM. The combined organic layers were washed with sat. aq. NaCl and dried over Na2SO4. The solvent was evaporated under vacuum and the residue was purified by rapid chromatography on a Si column by elution with DCM / EtOAc (1:1) to provide the title product (105 mg).

[0418] LCMS (Method 1): Rt = 1.20 min, ES + m / z 501.1 / 503.1 [M+H] + .

[0419] Step 3

[0420]

[0421] 7-(6-chloro-3-(3-(2-(dimethylamino)ethyl)-2-oxomylidene-1-yl)-1 H -pyrazolo[4,3- c ]pyridin-1-yl)-6-methoxy-2,3-dihydro-4 H -benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate 12)

[0422] Intermediate 12-2 (40.0 mg, 0.08 mmol), 2-bromo-N,N-dimethyl-ethylamine hydrobromide (22.3 mg, 0.10 mmol), and KO t A solution of Bu (21.5 mg, 0.19 mmol) in dry THF (2 mL) was stirred overnight at RT under nitrogen. RM was partitioned between DCM and water, and the aqueous layer was separately extracted with DCM. The combined organic layers were washed with sat. aq. NaCl, dried over Na2SO4, and the solvent was evaporated under vacuum. The residue was purified by rapid chromatography on a Si column by elution with DCM:MeOH (20.1 to 10:1) to provide the title product (34.5 mg).

[0423] LCMS (Method 1): Rt = 0.94 min, ES + m / z 572.3.1 / 574.2 [M+H] + .

[0424] Preparation of Examples

[0425] Example 1

[0426]

[0427] 6-Methoxy-7-(3-(3-methyl-2-oxomylidene-1-yl)-6-(pyrazolo[1,5-a]pyrimidine-3- (base)-1 H -pyrazolo[4,3- c ]pyridin-1-yl)-2 H -benzo[ b [1,4]Thiazine-3(4 H )-Ketone (Example 1)

[0428] A degassed mixture of intermediate 8b (9.0 mg, 20 µmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborpentane-2-yl)pyrazolo[1,5-a]pyrimidine (6.5 mg, 26 µmol), XPhos Pd G3 (2.6 mg, 3 µmol), and K3PO4 (8.6 mg, 40 µmol) in THF / water (3:1, 1.6 mL) was stirred at 70 °C for 1 h 45 min. After cooling to RT, RM was partitioned between DCM and water. The aqueous phase was further extracted with DCM (x2). The combined organic layers were dried over Na2SO4 and the solvent was removed under vacuum. The residue was purified by rapid chromatography on a Si column by elution with DCM / MeOH (20:1 to 10:1) to provide the title product (7 mg).

[0429] LCMS (Method 3), Rt = 4.94 min, ES + m / z 514.2 [M+H] +

[0430] 1 H-NMR (500 MHz, DMSO-d 6 ) δ: 9.63-9.69 (m, 1H), 9.18-9.25 (m, 1H), 8.87 (s, 1H), 8.69-8.75 (m, 1H), 8.05 (s, 1H), 7.15 (dd, J=7.0, 4.3 Hz, 1H), 6.95 (s, 1H), 6.56 (br s, 1H), 6.42 (s, 1H), 3.93 (br t, J=7.9 Hz, 2H), 3.64(s, 3H), 3.55-3.62 (m, 4H), 2.99 (br dd, J=5.3, 3.8 Hz, 2H), 2.87 (s, 3H).

[0431] Example 2

[0432]

[0433] 1-(1-(6-methoxy-3,4-dihydro-2) H -benzo[ b [1,4]thiazine-7-yl)-6-(pyrazolo[1,5-a]pyrimidine (Pyridine-3-yl)-1 H -pyrazolo[4,3- c ]Pyridin-3-yl)-3-methylimidazolidine-2-one (Example 2)

[0434] The title compound was prepared starting from intermediate 8c in a manner similar to that of Example 1.

[0435] LCMS (Method 3), Rt = 4.33 min, ES + m / z 528.2 [M+H] +

[0436] 1 H-NMR (500 MHz, DMSO-d 6 ) δ: 10.70-10.77 (m, 1H), 9.70 (s, 1H), 9.22(d, J=7.0 Hz, 1H), 8.88 (s, 1H), 8.71 (dd, J=4.0,1.5 Hz, 1H), 8.12 (s, 1H),7.48 (s, 1H), 7.16 (dd, J=6.9, 4.1 Hz, 1H), 6.96 (s, 1H), 3.96 (t, J=7.8 Hz,2H), 3.77 (s, 3H), 3.59 (t, J=7.9 Hz, 2H), 3.54 (s, 2H), 2.88 (s, 3H).

[0437] Example 3

[0438] Step 1

[0439]

[0440] 6-Methoxy-7-(3-(2-oxomylidene-3-yl)-6-(pyrazolo[1,5-) a ]pyrimidin-3-yl)-1 H -pyr Azo[4,3-] c ]pyridin-1-yl)-2,3-dihydro-4 H -benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate example) 3-1)

[0441] Starting with intermediate 5, the title compound was prepared in a manner similar to that of Example 1.

[0442] LCMS (Method 1): Rt = 1.14 min, ES+ m / z 585.1 [M+H] + .

[0443] Step 2

[0444]

[0445] 3-(1-(6-methoxy-3,4-dihydro-2) H -benzo[ b [1,4]oxazine-7-yl)-6-(pyrazolo[1,5-] a ]pyrethrum (Pyridine-3-yl)-1 H -pyrazolo[4,3- c ]Pyridin-3-yl)oxazolidin-2-one (Example 3)

[0446] The solution of intermediate Example 3-1 (33 mg, 56 µmol) and TFA (0.52 mL, 7.0 mmol) in DCM (3.8 mL) was stirred overnight at RT. RM was quenched with sat. aq. NaHCO3 and the aqueous layer was further extracted with DCM (3 × 20 mL). The combined organic layers were washed with sat. aq. NaHCO3 and sat. aq. NaCl, dried over Na2SO4, and the solvent was removed under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0-100% DCM / MeOH / NH4OH (90:5:0.5) in DCM to provide the title product (12.7 mg).

[0447] LCMS (Method 3), Rt = 3.42 min, ES + m / z 485.2 [M+H] +

[0448] 1 H-NMR (600 MHz, DMSO-d 6 ) δ: 9.55 (s, 1H), 9.22 (d, J=7.0 Hz, 1H), 8.89 (s, 1H), 8.72 (d, J=4.0, 1H), 8.10 (s, 1H), 7.16 (dd, J=7.0, 4.0 Hz, 1H), 6.77 (s, 1H), 6.48 (s, 1H), 6.31 (bs, 1H), 4.61 (t, J=7.9 Hz, 2H), 4.20(t, J=7.9 Hz, 2H), 4.13 (t, J=4.0 Hz, 2H), 3.64 (s, 3H), 3.38 (bs, 2H).

[0449] Examples 4 to 9

[0450] The following examples were prepared from the specified starting materials using a two-step method similar to that of Example 3.

[0451] Step 1

[0452]

[0453]

[0454] Step 2

[0455]

[0456]

[0457]

[0458]

[0459] Example 10

[0460] Step 1

[0461]

[0462] 7-(6-chloro-3-(5-(methoxycarbonyl)-2-oxomylidene-3-yl)-1 H -pyrazolo[4,3- c ]pyridine- 1-yl)-6-methoxy-2,3-dihydro-4 H -benzo[ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate Example 10-1)

[0463] The title product was prepared from intermediate 9-2 and methyl ethylene oxide-2-carboxylate in a manner similar to that of intermediate 9-3.

[0464] LCMS (Method 1): Rt = 1.28 min, ES + m / z 560.2 / 561.9 [M+H] + .

[0465] Step 2

[0466]

[0467] 3-(1-(4-(tert-butoxycarbonyl)-6-methoxy-3,4-dihydro-2 H -benzo[ b [1,4]oxazine-7-yl)-6- (pyrazolo[1,5- a ]pyrimidin-3-yl)-1 H -pyrazolo[4,3- c ]pyridin-3-yl)-2-oxomylidene-5-carboxylic acid (middle) Intermediate Example 10-2)

[0468] Starting with intermediate example 10-1, in order to... Example 1 The title product was prepared in a similar manner.

[0469] LCMS (Method 1): Rt = 0.88 min, ES + m / z 629.2 [M+H] + .

[0470] Step 3

[0471]

[0472] 7-(3-(5-(ethoxycarbonyl)-2-oxylidene-3-yl)-6-(pyrazolo[1,5-) a ]pyrimidine-3-yl)- 1 H -pyrazolo[4,3-c]pyridin-1-yl)-6-methoxy-2,3-dihydro-4 H -benzo[ b [1,4]oxazine-4-carboxylic acid tert-butyl ester (Intermediate Examples 10-3)

[0473] The mixture of intermediate Example 10-2 (30 mg, 0.05 mmol), iodoethane (11 mg, 0.07 mmol), and K₂CO₃ (6.6 mg, 0.05 mmol) in DMF (1.0 mL) was stirred at RT for 3 h. RM was diluted with water and extracted with EtOAc (4 × 15 mL). The combined organic layers were washed with sat. aq. NaCl, dried over Na₂SO₄, and the solvent was evaporated under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0–90% DCM / MeOH / NH₄OH (90:9:0.5) in DCM to provide the desired product (6.3 mg).

[0474] LCMS (Method 1): Rt = 1.05 min, ES + m / z 657.2 [M+H] + .

[0475] Step 4

[0476]

[0477] 3-(1-(6-methoxy-3,4-dihydro-2) H -benzo[ b [1,4]oxazine-7-yl)-6-(pyrazolo[1,5-] a ]pyrethrum (Pyridine-3-yl)-1 H -pyrazolo[4,3- c ethyl pyridin-3-yl)-2-oxomylidene-5-carboxylate (Example) 10)

[0478] Starting with intermediate examples 10-3, to be consistent with Step 2 of Example 3 The title product was prepared in a similar manner.

[0479] LCMS (Method 3), Rt = 4.91 min, ES + m / z 557.2 [M+H] +

[0480] 1 H-NMR (600 MHz, DMSO-d 6) δ: 9.55 (d, J=1.1 Hz, 1H), 9.23 (dd, J=7.0,1.7 Hz, 1H), 8.89 (s, 1H), 8.73 (dd, J=4.0, 1.7 Hz, 1H), 8.12 (d, J=1.1 Hz,1H), 7.17 (dd, J=7.0, 4.0 Hz, 1H), 6.79 (s, 1H), 6.49 (s, 1H), 6.33-6.31 (m,1H), 5.47 (dd, J=9.7, 5.3 Hz, 1H), 4.51 (t, J=9.7 Hz, 1H), 4.30-4-23 (m, 3H),4.14 (t, J=4.3 Hz, 2H), 3.64 (s, 3H), 3.40-3-37 (m, 2H, overlapping with HDO), 1.27 (t, J=7.0 Hz, 3H).

[0481] Examples 11 and 12

[0482]

[0483] ( S )-5-((dimethylamino)methyl)-3-(1-(6-methoxy-3,4-dihydro-2 H- Benzo[ b [1,4]Oxazine- 7-yl)-6-(pyrazolo[1,5) -a ]pyrimidin-3-yl)-1 H- Pyrazolo[4,3] -c ]pyridin-3-yl)oxazolidin-2-one (Example) 11) and ( R )-5-((dimethylamino)methyl)-3-(1-(6-methoxy-3,4-dihydro-2 H- Benzo[ b [1,4]Oxazine-7- )-6-(pyrazolo[1,5-) -a ]pyrimidin-3-yl)-1 H- Pyrazolo[4,3] -c ]pyridin-3-yl)oxazolidin-2-one (Example) 12)

[0484] Examples 11 and 12 were obtained by chiral separation from the (S)-rich partially racemic mixture of Example 4 (126 mg) using chiral chromatography. The samples were dissolved in acetonitrile and purified on a Lux C4 column (21.5 mm x 250 mm, 5 μm) under RT, isocratic conditions (MeOH), a flow rate of 21 mL / min, a detector at 220 nm, and an injection volume of 2000 μL to provide Examples 11 (55 mg) and 12 (12 mg).

[0485] Chiral purity analysis method

[0486] LuxC2 column (4.6 mm x 150 mm, 5 μm) was used at ambient temperature, isocratic conditions (MeOH), flow rate 1 mL / min, detector in 210–400 nm, and injection volume 1 μL.

[0487] Example 11

[0488] LCMS (Method 3), Rt = 3.40 min, ES + m / z542.2 [M+H] +

[0489] Chiral chromatography (chiral purity analysis method): Rt = 13.01 min, ee = 100.0%

[0490] 1 H-NMR (500 MHz, DMSO-d 6 ) δ: 9.57 (s, 1H), 9.23 (dd, J=6.9, 1.4 Hz,1H), 8.89 (s, 1H), 8.72 (dd, J=4.0, 1.5 Hz, 1H), 8.10 (s, 1H), 7.16 (dd, J=6.9, 4.1 Hz, 1H), 6.77 (s, 1H), 6.47 (s, 1H), 6.31 (s, 1H), 4.98 (quin, J=7.1Hz, 1H), 4.25 (t, J=9.0 Hz, 1H), 4.13 (t, J=4.1 Hz, 2H), 3.90 (dd, J=9.1,7.1Hz, 1H), 3.64 (s, 3H), 3.35-3.40 (m, 2H), 2.69 (dd, J=13.3,6.2 Hz, 1H), 2.66 (dd, J=13.3,5.0 Hz, 1H), 2.26 (s, 6H).

[0491] Example 12

[0492] LCMS (Method 3), Rt = 3.41 min, ES + m / z 542.2 [M+H] +

[0493] Chiral chromatography (chiral purity analysis method): Rt = 19.99 min, ee = 99.2%

[0494] 1 H-NMR (600 MHz, DMSO-d 6) δ: 9.57 (s, 1H), 9.23 (dd, J=6.9, 1.4 Hz,1H), 8.89 (s, 1H), 8.72 (dd, J=4.0, 1.5 Hz, 1H), 8.10 (s, 1H), 7.16 (dd, J=6.9, 4.1 Hz, 1H), 6.77 (s, 1H), 6.47 (s, 1H), 6.31 (s, 1H), 4.98 (quin, J=7.1Hz, 1H), 4.25 (t, J=9.0 Hz, 1H), 4.13 (t, J=4.1 Hz, 2H), 3.90 (dd, J=9.1,7.1Hz, 1H), 3.64 (s, 3H), 3.35-3.40 (m, 2H), 2.69 (dd, J=13.3,6.2 Hz, 1H), 2.66 (dd, J=13.3,5.0 Hz, 1H), 2.26 (s, 6H).

[0495] Chirality specification

[0496] The synthesis of the (S)-rich semi-racemic mixture of Example 4 was carried out from the (R)-rich semi-racemic mixture of 5-(chloromethyl)oxazolidin-2-one (ee% = 60%) according to the procedure of Example 4. Assuming that no racemization occurred during the conversion from semi-racemic 5-(chloromethyl)oxazolidin-2-one to semi-racemic intermediate 4a, and from semi-racemic intermediate 4a to semi-racemic Example 4, the major separation peak during chiral separation was designated as the product derived from (R)-5-(chloromethyl)oxazolidin-2-one (Example 11).

[0497] Example 13

[0498] Step 1

[0499]

[0500] ( R )-7-(3-(5-((dimethylamino)methyl)-2-oxadiazolidin-3-yl)-6-((3-methoxypyrazine- 2-yl)amino)-1 H- Pyrazolo[4,3] -c ]pyridin-1-yl)-6-methoxy-2,3-dihydro-4 H- Benzo[ b [1,4]Oxazine- 4-Tert-butyl carbamate (intermediate, Example 13-1)

[0501] The degassed mixture of intermediate 6b (300 mg, 0.54 mmol), 2-amino-3-methoxypyrazine (87 mg, 0.70 mmol), sodium tert-butoxide (77 mg, 0.80 mmol), and RuPhos Pd G3 (67 mg, 80 μmol) in dioxane (11 mL) was stirred at 85 °C for 1.5 h. After cooling to RT, the solvent was removed under vacuum, and the residue was purified by rapid chromatography on a Si column by elution with 0–50% DCM / MeOH / NH4OH (90:15:1.5) in DCM to provide the title product (220 mg).

[0502] LCMS (Method 2), Rt = 1.32, ES + m / z 648.4 [M+H] +

[0503] Step 2

[0504]

[0505] ( R )-5-((dimethylamino)methyl)-3-(1-(6-methoxy-3,4-dihydro-2 H- Benzo[ b [1,4]Oxazine- 7-yl)-6-((3-oxoylide-3,4-dihydropyrazin-2-yl)amino)-1 H- Pyrazolo[4,3] -c ]pyridin-3-yl)oxazolidine- 2-Ketone (Example 13)

[0506] The suspension of intermediate Example 13-1 (220 mg, 0.30 mmol) in MeCN (5.44 mL) was treated with NaI (137 mg, 0.92 mmol) and TMS-Cl (116 µL, 0.92 mmol). The solvent was stirred at 85 °C for 4 h. After cooling to RT, the solvent was removed under vacuum. The residue was purified by rapid chromatography on a Si column by elution with 0-100% DCM / MeOH / NH4OH (90:15:1.5) in DCM to provide the title product (150 mg).

[0507] LCMS (Method 3), Rt = 3.18 min, ES + m / z 534.1 [M+H] +

[0508] 1 H-NMR (600 MHz, DMSO - d 6) δ: 12.20 (br.s, 1H), 9.29 (d, J=1.0 Hz, 1H), 8.73 (s, 1H), 8.00 (d, J=1.0 Hz, 1H), 6.93 (d, J=4.4 Hz, 1H), 6.89 (d, J=4.4Hz, 1H), 6.72 (s, 1H), 6.45 (s, 1H), 6.28 (br.s., 1H), 4.99-4.93 (m, 1H), 4.21 (t, J=9.0 Hz, 1H), 4.12 (t, J=4.3 Hz, 2H), 3.87 (dd, J=9.0, 7.1 Hz, 1H), 3.63(s, 3H), 3.38–3.32 (m, 2H, overlapping with HDO), 2.69–2.62 (m, 2H), 2.25 (s, 6H).

[0509] Example 14

[0510] Step 1

[0511]

[0512] ( S )-6-methoxy-7-(3-(5-(((2-methoxyethyl)(methyl)amino)methyl)-2-oxadiazolidine- 3-yl)-6-((3-methoxypyrazin-2-yl)amino)-1 H -pyrazolo[4,3- c ]pyridin-1-yl)-2,3-dihydro-4 H -Benzo [ b [1,4]Oxazine-4-carboxylic acid tert-butyl ester (intermediate Example 14-1)

[0513] Starting with intermediate 10, the title compound was prepared in a manner similar to step 1 of Example 13.

[0514] LCMS (Method 2), Rt = 1.35, ES + m / z 692.3 [M+H] +

[0515] Step 2

[0516]

[0517] ( S )-3-(1-(6-methoxy-3,4-dihydro-2 H -benzo[ b [1,4]Oxazine-7-yl)-6-((3-oxo-3, 4-Dihydropyrazine-2-yl)amino)-1 H -pyrazolo[4,3- c ]pyridin-3-yl)-5-(((2-methoxyethyl)(methyl)amino (Methyl)oxazolidin-2-one (Example 14)

[0518] Starting with intermediate Example 14-1, the title compound was prepared in a manner similar to step 2 of Example 13.

[0519] LCMS (Method 3), Rt = 3.39 min, ES + m / z 578.1 [M+H] +

[0520] 1 H-NMR (500 MHz, DMSO - d 6 ) δ: 12.18 (br.s, 1H), 9.30 (d, J=0.8 Hz, 1H), 8.74 (s, 1H), 8.00 (d, J=0.8 Hz, 1H), 6.93 (d, J=4.4 Hz, 1H), 6.87 (d, J=4.4Hz, 1H), 6.72 (s, 1H), 6.45 (s, 1H), 6.28 (br.s, 1H), 4.97-4.91 (m, 1H), 4.19(t, J=9.2 Hz, 1H), 4.12 (t, J=4.1 Hz, 2H), 3.89 (dd, J= 9.2, 7.0 Hz, 1H),3.63 (s, 3H), 3.42 (t, J=5.8 Hz, 2H), 3.35 (m, 2H), 3.21 (s, 3H), 2.79 (d, J=5.6 Hz, 2H), 2.61 (t, J=5.8 Hz, 2H), 2.31 (s, 3H).

[0521] Pharmacological activities of compounds (1-14) of the present invention

[0522] Biochemical titers of JAK1, JAK2, JAK3 and Tyk2

[0523] Measurement principle

[0524] The aim of this study was to evaluate the ability of compounds to inhibit the activity of all four JAK isoforms in a cell-free environment. JAK 1, JAK 2, JAK 3, and TYK2 were determined using time-resolved fluorescence resonance energy transfer (TR-FRET) technology. This technique involves measuring the interaction between two labeled binding partners by measuring energy transfer from the excited donor to the acceptor dye and the light emission of the acceptor dye. The LANCE Ultra kinase assay was used. In the presence of JAK 1, JAK 2, JAK 3, and TYK2 kinases and ATP (equivalent to Km), the ULight peptide substrate (LANCE Ulight-JAK-1 (Tyr1023) peptide, Perkin Elmer, TRF0121) was phosphorylated. It was then captured by an Eu-anti-phospho-substrate antibody (LANCEEu-W1024 antiphosphotyrosine (PT66), Perkin Elmer, AD0069), which brought the Eu-chelate donor and the ULight acceptor dye into close proximity. Upon excitation at 320 nm, the Eu-chelate transfers its energy to the ULight dye, resulting in fluorescence emission at 665 nm.

[0525] Compound testing

[0526] A series of dilutions of the compounds in pure DMSO were prepared from a 10 mM DMSO stock solution. Eleven consecutive 5-fold dilutions (20 µM – 2 pM) of the compounds were tested in 384-well plates, starting from a maximum concentration of 20 µM. 200 nL of the compound was transferred from the master plate to the test plate using a Mosquito (TTP Labtech) apparatus. Assays were performed in 384-well Perkin Elmer test plates with a 20 μL assay volume (kinase reaction) and a total volume of 40 μL (stop reagent and antibody assay). For JAK 1, JAK 2, JAK 3, and TYK2, 30 / 50 / 20 / 10 nM peptide and 20 / 0.7 / 0.2 / 12 µM ATP were added to 10 μL of substrate solution (peptide + ATP). 10 μL of enzyme solution was added to the kinase reaction at the following concentrations: 0.15 / 0.083 / 0.025 / 0.144 ng / µL of JAK 1, JAK 2, JAK 3, and TYK2, respectively. After shaking and incubation at room temperature (rt) for 1.5 h, 20 μL of stop (10 μL EDTA) and the assay mixture (10 μL europium-anti-phosphorylation antibody, final: 0.5 nM) were added. Readings were taken on an EnVision 2104 reader (Perkin Elmer) after 1 h of incubation.

[0527] IC50 data, curves, and QC analysis were calculated using Excel and GraphPadPrism software v9. In short, individual concentration-response curves were generated by plotting the logarithm (X) of the test concentration of the test compound against the corresponding percentage of inhibition (Y) using a least-squares (ordinary) fit. The best-fit IC50 value was calculated using the Log(inhibitor) versus the normalized response-variable slope equation, where Y = 100 / (1 + 10^(LogIC50 - X)). Hill slope). The QC standard parameters (Z', S:B, R², Hill slope) for each IC50 curve were checked. IC50 data, curves, and QC analysis were calculated using Excel and GraphPadPrism software. QC standard parameters: Z' ≥ 0.5, Hill slope range 0.5 to 5, S:B > 2.

[0528] The compounds according to the invention exhibit pIC50 values ​​greater than 6 in terms of their inhibitory activity against all JAK isoforms, which corresponds to an inhibitory concentration of ≤ 1 µM. Most compounds preferably exhibit pIC50 values ​​greater than 7.7, and even more preferably greater than 8.7, in terms of their inhibitory activity against JAK1; which corresponds to an inhibitory concentration of ≤20 nM, and even more preferably ≤2 nM.

[0529] The data for compounds 1-12 are reported in the table below.

[0530]

[0531] Based on the following classification criteria, the compounds are classified in the table above according to their potency against the inhibitory activities of JAK1, JAK2, JAK3, and TYK2 isoforms:

[0532] + + +:pIC 50 ≥ 8.7

[0533] ++:8.7 > pIC 50 ≥ 7.7

[0534] +:pIC 50 < 7.7

[0535] Inhibition of pSTAT6 induced by IL-13 in BEAS

[0536] BEAS-2B human cell line (100,000 cells / well) was seeded and incubated at 37°C, 5% CO2, and 95% humidity for 48 h. The compound was added and incubated for 30 min, followed by IL-13 as a trigger. After 30 min incubation, cells were lysed and pSTAT6 was measured using a Fastscan phospho-stat6 (Tyr641) sandwich ELISA kit (cell signaling). Inhibitors were detected in duplicate at 11 consecutive 5-fold dilutions (10 µM – 40 pM) starting at 10 μM. IC50 data, curves, and QC analysis were calculated using Excel and GraphPadPrism software. QC criteria: Z' ≥ 0.35, Hill slope range 0.5 to 5, S:B > 2.

[0537] The compounds according to the invention exhibit measurable values ​​above 5.3 in terms of pIC50 (BEAS).

[0538]

[0539] Based on the following classification criteria, the compounds are classified in the table above according to their potency in terms of functional activity in relation to BEAS:

[0540] §§§§:pIC 50 ≥ 8.3

[0541] §§§:8.3 > pIC 50 ≥ 7.3

[0542] §§:7.3 > pIC 50 ≥ 6.3

[0543] §:pIC 50 < 6.3

[0544] If a numerical limit or range is stated in this document, the endpoints are included. Furthermore, when not explicitly stated, all values ​​and subranges within the numerical limit or range are included.

[0545] As used in this article, the words “a” and “a kind” have the meaning of “one / kind or multiple / kinds”.

[0546] Obviously, many variations and modifications of the invention are possible based on the above teachings. Therefore, it should be understood that the invention can be practiced in ways other than those specifically described herein, within the scope of the appended claims.

Claims

1. Compounds of formula (I) in R1 is selected from the following heteroaryl groups. ； R2 is selected from and in K is selected from O and S; R3 is a monocyclic urea or carbamate group selected from the following formula J. R7 is selected from H, (C1-C6)alkyl, (C1-C6)hydroxyalkyl, and -(CH2). m NR4R5, (C1-C6)alkoxycarbonyl (CH2) m , in, m is independently 0 or an integer from 1 to 4 each time it appears; R4 and R5 may be the same or different, and are independently selected from -H, (C1-C6)alkyl, (C1-C6)alkoxy-(C1-C6)alkyl; Its single enantiomers, diastereomers, and mixtures, Or its pharmaceutically acceptable salts or solvates.

2. The compound of formula (I) according to claim 1, wherein the compound is represented by general formula (Ib): in R3 is J2; and R7 is selected from H, (C1-C6) alkyl, and -(CH2). m NR4R5, Its single enantiomers, diastereomers, and mixtures, And its pharmaceutically acceptable salts and solvates.

3. The compound of formula (I) according to claim 1, wherein the compound is represented by general formula (Ic): in R3 is J2; and R7 is selected from H, (C1-C6) alkyl, and -(CH2). m NR4R5, Its single enantiomers, diastereomers and mixtures, or its pharmaceutically acceptable salts or solvates.

4. The compound according to claim 1, wherein the compound is selected from the following list: 6-Methoxy-7-(3-(3-methyl-2-oxomimidazolidine-1-yl)-6-(pyrazolo[1,5-a]pyrimidin-3-yl)-1H-pyrazolo[4,3-c]pyridin-1-yl)-2H-benzo[b][1,4]thiazin-3(4H)-one; 1-(1-(6-methoxy-3,4-dihydro-2H-benzo[b][1,4]thiazin-7-yl)-6-(pyrazolo[1,5-a]pyrimidin-3-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)-3-methylimidazolidine-2-one; 3-(1-(6-methoxy-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-6-(pyrazolo[1,5-a]pyrimidin-3-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)oxazolidin-2-one; 5-((dimethylamino)methyl)-3-(1-(6-methoxy-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-6-(pyrazolo[1,5-a]pyrimidin-3-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)oxazolidin-2-one; 3-(1-(6-methoxy-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-6-(pyrazolo[1,5-a]pyrimidin-3-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)-5-((methylamino)methyl)oxazolidin-2-one; 5-(2-(dimethylamino)ethyl)-3-(1-(6-methoxy-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-6-(pyrazolo[1,5-a]pyrimidin-3-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)oxazolidin-2-one; (S)-5-(hydroxymethyl)-3-(1-(6-methoxy-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-6-(pyrazolo[1,5-a]pyrimidin-3-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)oxazolidin-2-one; 1-(1-(6-methoxy-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-6-(pyrazolo[1,5-a]pyrimidin-3-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)-3-methylimidazolidine-2-one; 1-(2-(dimethylamino)ethyl)-3-(1-(6-methoxy-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-6-(pyrazolo[1,5-a]pyrimidin-3-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)imidazolidine-2-one; 3-(1-(6-methoxy-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-6-(pyrazolo[1,5-a]pyrimidin-3-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)-2-oxomylidene-5-carboxylic acid ethyl ester; (S)-5-((dimethylamino)methyl)-3-(1-(6-methoxy-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-6-(pyrazolo[1,5-a]pyrimidin-3-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)oxazolidin-2-one; (R)-5-((dimethylamino)methyl)-3-(1-(6-methoxy-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-6-(pyrazolo[1,5-a]pyrimidin-3-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)oxazolidin-2-one; (R)-5-((dimethylamino)methyl)-3-(1-(6-methoxy-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-6-((3-oxoylide-3,4-dihydropyrazin-2-yl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)oxazolidin-2-one; (S)-3-(1-(6-methoxy-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-6-((3-oxoylide-3,4-dihydropyrazin-2-yl)amino)-1H-pyrazolo[4,3-c]pyridin-3-yl)-5-(((2-methoxyethyl)(methyl)amino)methyl)oxazolidin-2-one; Its single enantiomers, diastereomers and mixtures, or pharmaceutically acceptable salts and solvates thereof.

5. A pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof as defined in any one of claims 1 to 4, mixed with one or more pharmaceutically acceptable carriers or excipients.

6. The pharmaceutical composition of claim 5, which is suitable for administration by inhalation and is selected from inhalable powders, metered aerosols containing propellants, or inhalable formulations without propellants.

7. An apparatus comprising the pharmaceutical composition of claim 6, which may be a single-dose or multi-dose dry powder inhaler, a metered-dose inhaler, or a soft-nebulizer.

8. The compound or pharmaceutical composition according to any one of claims 1 to 5, used as a medicine.

9. The compound or pharmaceutical composition of claim 8 for the stated use, for the prevention and / or treatment of lung diseases selected from asthma, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), acute lung injury, and acute respiratory distress syndrome (ARDS).

10. A combination of a compound as defined in any one of claims 1 to 4 with one or more active ingredients, said active ingredient being selected from the classes currently used to treat respiratory disorders and classes known to those skilled in the art, such as β2-agonists, antimuscarinic drugs, corticosteroids, mitogen-activated kinase (P38 MAP kinase) inhibitors, PI3K inhibitors (phosphoinositol 3-kinase), nuclear factor κ-B kinase β subunit inhibitors (IKK2), Rho kinase inhibitors (ROCKi), human neutrophil elastase (HNE) inhibitors, phosphodiesterase 4 (PDE4) inhibitors, leukotriene modifiers, nonsteroidal anti-inflammatory drugs (NSAIDs), and mucus modifiers.

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