RU2833004C2 - Heterocyclic pyrazole derivatives as type iii receptor tyrosikinase inhibitors - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula (II), (III), (IV) or (V), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. In the above formulas, the values of the variables are as follows: G is selected from a group consisting of an optionally substituted C6-aryl, optionally substituted 5–6 membered heteroaryl having one or more heteroatoms selected from N and S, optionally substituted 5–8 membered heterocyclyl having one or more heteroatoms selected from O and N, C2-C10 alkynyl and a direct bond; L is selected from a group consisting of –CH=, -CHR⁴-, -(CH₂)_q-, -NR⁵-, -O-, -O(CH₂)_q-, -C(O)-, -C(O)(CH₂)_q-, -SO₂- and direct communication, in which each q is individually and independently integer from 1 to 4; Z is selected from the group consisting of hydrogen, deuterium, halogen, C1-C4 alkyl, C1-C4 alkylamine, C1-C4 dialkylamine and optionally substituted 4–7 membered heterocyclyl or a heterocyclic spiro compound; R¹ is selected from a group consisting of hydrogen and deuterium; R² is selected from the group consisting of hydrogen, deuterium, halogen and C1-C6 alkyl; R³ is selected from the group consisting of hydrogen, deuterium and C1-C6 alkyl; R⁴ is selected from the group consisting of deuterium and C1-C6 alkyl; and R⁵ is selected from the group consisting of hydrogen and C1-C6 alkyl. Also disclosed is a pharmaceutical composition containing said compound, and a method of treating using said compound.

EFFECT: disclosed compounds inhibit RTK class III kinases, such as kinases FLT3, PDGFR, and can be used to treat diseases or disorders mediated by such kinases, particularly cancer.

10 cl, 7 tbl, 6 ex

Description

Field of technology to which the invention relates

The present invention relates to new chemical compounds and to methods of using them in therapy and preparations. In particular, the invention relates to certain substituted heterocyclic pyrazole compounds and to methods of using them in inhibiting, regulating and/or modulating type III receptor tyrosine kinases such as FLT3, PDGFR, c-KIT and/or CSF-1R and their associated signal transduction pathways.

State of the art

Protein kinases (PKs) play important roles in cellular signal transduction pathways that regulate various cellular functions such as differentiation, proliferation, migration, survival, and apoptosis. These enzymes catalyze the transfer of a phosphate group from ATP to a tyrosine, serine, or threonine residue on a protein substrate. Kinase phosphorylation and phosphatase dephosphorylation are involved in countless cellular processes that respond to various intracellular or extracellular signals, regulation of cellular functions, and activation or deactivation of cellular operations. Receptor tyrosine kinases (RTKs) are a subfamily of protein kinases, and a class of RTKs known as the receptor tyrosine kinase type III family, which includes FLT3, PDGFR, c-KIT, and CSF-1R, has been associated with various proliferative and inflammatory diseases.

Small molecule inhibitors of receptor tyrosine kinase type III provide a rational approach to new therapies for the treatment of autoimmune diseases and, in particular, to blocking chronic tissue destruction mediated by the innate immune system. Moreover, inhibition of receptor tyrosine kinase type III also provides a rational approach to new therapies for the treatment of cancer, especially for the treatment of cancer invasiveness, cancer angiogenesis or vasculogenesis, cancer metastasis, and cancer immunotolerance.

FLT3 (Fms-like tyrosine kinase 3, also called Flk2) and its ligand (FL) are one of the regulators of normal hematopoiesis, and activating mutation or overexpression of FLT3 is common in AML. More than a dozen known FLT3 inhibitors are under development, some of which have shown promising clinical effects and have received approval for the treatment of FLT3 mutant-positive AML. The FLT3 receptor is also expressed on a significant proportion of dendritic cell precursors, and stimulation of the FLT3 receptor induces proliferation and differentiation of these precursors into dendritic cells (DCs). Because dendritic cells are the primary initiators of T-cell immune responses, including autoreactive immune responses, FLT3 inhibition provides a mechanism to downregulate DC-mediated inflammatory or autoimmune responses. One study shows that the FLT3 inhibitor CEP-701 is effective in reducing myelin loss in experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis. High levels of FLT3 ligand are found in the serum of patients with Langerhans cell histiocytosis and systemic lupus erythematosus. This observation further implicates activation of the FLT3 pathway in the dysregulation of dendritic cell precursors in these autoimmune diseases.

PDGFR (platelet-derived growth factor receptor) and its ligand (PDGF) regulate cell proliferation, survival, differentiation and migration of cells, primarily of mesenchymal origin. Abnormalities in the PDGF pathway, including overexpression or enhancement of PDGF receptors (PDGFRs), amplification of hotspot mutations or activating chromosomal translocations, have been observed in a wide range of pathological conditions such as cancer, fibrosis, neurological conditions and atherosclerosis, making them potential targets for disease treatment.

c-KIT (also known as stem cell growth factor receptor, SCFR) and its ligand (SCF) induce proliferation, differentiation, or migration of cells within the hematopoietic, hematogenous, and melanogenic lineages at various stages of development. The presence of c-KIT mutations is a key diagnostic marker for gastrointestinal stromal tumor (GIST). Gleevec (imatinib mesylate or STI571), the first FDA-approved receptor tyrosine kinase (RTK) inhibitor, initially approved for the treatment of c-Abl-mediated chronic myeloid leukemia, received FDA approval for c-KIT-mediated GIST in 2002 and validated the molecular approach of c-KIT inhibition for the treatment of GIST. Gain-of-function mutations in c-KIT are also associated with mast cell/myeloid leukemia and seminomas/dysgerminomas. c-KIT mutations have also been identified in some melanomas and have been identified as a potential therapeutic target for melanoma.

CSF1R (also known as macrophage colony-stimulating factor receptor (M-CSFR) or Fms) is the receptor for macrophage colony-stimulating factor (M-CSF or CSF-1). CSF-1/CSF-1R is the primary growth factor that regulates the survival, proliferation, and differentiation of cells of the mononuclear phagocytic lineage. Numerous studies have demonstrated that CSF-1/CSF-1R plays a role in tumor tissues. Overexpression or activation of CSF-1R and/or its ligand has been detected in a variety of cancers, and elevated M-CSF levels are associated with poor prognosis in some cancers. M-CSF is one of several cytokines involved in the selection of tumor-associated macrophages (TAMs), which promote tumor angiogenesis and tumor progression to metastasis. Activation of CSF-1R also leads to proliferation and differentiation of osteoclast precursors, thereby mediating the process of bone resorption. Therefore, inhibition of CSF-1R provides treatment for cancer, especially invasion, angiogenesis, cancer metastasis, cancer immunotolerance, and bone metastasis. Due to its role in osteoclast biology, CSF-1R is also an important therapeutic target for osteoporosis, inflammatory arthritis, and other inflammatory bone erosions.

Although various tyrosine kinase inhibitors have been shown to be useful therapeutic agents, there remains a need for type III kinase inhibitors.

Disclosure of invention

The present invention relates to novel inhibitors that inhibit kinases, especially class III RTKs, such as FLT3, PDGFRα, c-KIT and/or CSF-1R kinases, for the treatment of diseases or disorders mediated by such kinases. The inhibitors of the present invention also find use in the treatment of other mammalian diseases, including human diseases, mediated by FLT3, PDGFR, c-KIT or CSF-1R kinases. Such diseases include, but are not limited to, cancer, autoimmune diseases and bone resorptive diseases.

Embodiments of the invention are based on the unexpected findings that certain heterocyclic pyrazole compounds can inhibit the activity of members of the type III receptor tyrosine kinase family (e.g., FLT3, PDGFR, c-KIT, and CSF-1R). The compounds are useful in therapeutic methods, pharmaceutical compositions, and methods for modulating the activity of FLT3, PDGFR, c-KIT, and/or CSF-1R kinases, including wild-type and/or mutated forms of FLT3, PDGFR, c-KIT, and/or CSF-1R kinases. These properties enable the use of these heterocyclic pyrazole moieties in the treatment of diseases and/or conditions associated with protein tyrosine kinase.

According to embodiments of the present invention, the compound may have the general Formula I:

Formula (I)

or a stereoisomer, tautomer, pharmaceutically acceptable salt, proteolysis-targeted chimera (PROTAC) linker thereof, in which

X is selected from the group consisting of CR ² and NR ³ ;

Y is selected from the group consisting of CR ² and NR ³ ;

G is selected from the group consisting of optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, alkynyl, and direct bond;

L is selected from the group consisting of -CH=, ^-CHR4- , -( _CH2 ) _q- , ^-NR5- , -O-, -O( _CH2 ) _q- , -C(O)-, -C(O)( _CH2 ) _q- , -C(O)NH-, _-SO2- , and a direct bond, wherein q is individually or independently an integer from 1 to 4;

Z is selected from the group consisting of hydrogen, C1-C4 alkyl, C1-C4 alkylamine, C1-C4 dialkylamine, and optionally substituted heterocyclyl and heterocyclic spiro compound;

R ¹ is selected from the group consisting of hydrogen, deuterium, halogen, hydroxyl, amino, cyano, trifluoromethyl, trifluoromethoxy, nitro, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino and C1-C6 dialkylamino;

R ² is selected from the group consisting of hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C5-C6 cycloalkenyl, C1-C6 alkylamino, C1-C6 dialkylamino, C3-C6 cycloalkylamino, C1-C6 alkoxy, C3-C6 cycloalkoxy, aryl, 3-6-membered heterocyclyl, and 5-6-membered heteroaryl, wherein the alkylamino, dialkylamino, alkoxy, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkylamino, cycloalkoxy, heterocyclyl, aryl and heteroaryl are optionally substituted with halogen, amino, hydroxyl, cyano, nitro, acrylic, acyloxy, C1-C4 alkyl, C1-C4 hydroxyl alkyl, C1-C4 alkoxy alkyl, C1-C4 acyloxy alkyl, C1-C4 alkoxy, C1-C4 dialkylamino, C3-C6 cycloalkyl, 3-6-membered heterocyclyl, aryl and 5-6-membered heteroaryl;

R ³ is selected from the group consisting of hydrogen, deuterium, C1-C6 alkyl, C1-C6 alkyl carbonyl, C2-C6 alkenyl carbonyl, C1-C6 alkoxy carbonyl, amino carbonyl, C1-C6 alkylamino carbonyl and C1-C6 dialkylamino carbonyl; and

R ⁴ is selected from the group consisting of hydrogen, deuterium and C1-C6 alkyl; and

R ⁵ is selected from the group consisting of hydrogen and C ₁ -C ₆ alkyl.

In another aspect, embodiments of the invention provide pharmaceutical compositions, each comprising a compound of Formula I, a stereoisomer, tautomer, solvate, prodrug, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, or excipient.

In another aspect, embodiments of the invention provide methods of inhibiting type III receptor tyrosine kinases, such as FLT3, PDGFR, c-KIT and/or CSF-1R kinases in a mammal. The method of the invention comprises: administering to said mammal in need thereof a therapeutically effective amount of a compound of Formula I.

In another aspect of the invention, embodiments of the invention provide a proteolysis-targeted chimera binder (PROTAC) for the degradation of type III receptor tyrosine kinases, such as FLT3, PDGFR, c-KIT and/or CSF-1R kinases in mammals via the ubiquitin pathway. The proteolysis-targeted chimera (PROTAC), a type of dibasic molecule, can remove unwanted proteins induced by intracellular proteolytic selectivity. The PROTAC consists of two protein-binding portions, one for an E3 ubiquitin ligase and one for target proteins (the binder described herein). By binding the two proteins, the PROTAC introduces the target protein to the E3 ligase, resulting in the tagging (e.g., ubiquitination) of the target protein for subsequent degradation by the proteasome. Many molecules are known to bind various E3 ubiquitin ligases, such as nutlin, methyl-bestatin, VHL ligand, thalidomide, pomalidomide and lenalidomide, as described in WO 2019/140003 A1, the disclosure of which is incorporated by reference. Any of these E3 ligase binders can be combined with a compound of the invention. A linker can be used between the two linking moieties to provide spacing and ease of implementation of the coupling reactions. Some examples of linkers can be found in WO 2019/140003. In addition, Burslem et al. were inspired by the utility of receptor tyrosine kinases for the degradation of target proteins via proteolysis-targeted chimeras (PROTACs) at low nanomolar concentrations. They used a known FLT3 inhibitor, quizartinib, as a recruitment element and installed von Hippel-Lindau ligand (VHL) via an optimized linker to develop a FLT3 PROTAC. This compound efficiently induced FLT3-ITD protein degradation in MOLM-14 cells and MV4-11 cells at low nanomolar concentrations (Journal of the American Chemical Society 2018; 140: 16428-16432).

In another aspect, embodiments of the invention provide methods for treating a disease or disorder selected from fibrosis, a bone-related disease, cancer, an autoimmune disorder, an inflammatory disease, and a cardiovascular disease in a mammal. The method of the invention comprises: administering to said mammal in need thereof a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof. A method for treating a disease or disorder mediated by FLT3 kinases and/or PDGFR kinases or mediated by a mutant kinase of FLT3 kinases and/or PDGFR kinases is provided, comprising: administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition comprising the above compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable diluent or carrier, or a protein targeting module.

In another aspect, embodiments of the invention provide for the use of a compound of Formula I in the manufacture of a medicament for the treatment of a disease or disorder selected from fibrosis, a bone-related disease, cancer, an autoimmune disorder, an inflammatory disease, and a cardiovascular disease in a mammal.

In another aspect, embodiments of the invention provide for the use of a compound of Formula I in the treatment of a disease or disorder selected from fibrosis, a bone-related disease, cancer, an autoimmune disorder, an inflammatory disease, and a cardiovascular disease in a mammal.

In another aspect, embodiments of the invention provide intermediates for the preparation of compounds of Formula I. In one embodiment, certain compounds of Formula I can be used as intermediates for the preparation of other compounds of Formula I.

In another aspect, embodiments of the invention provide processes for preparing, methods for separating, and methods for purifying the compounds described herein.

Other aspects and advantages of the invention will become apparent from the following description and the appended claims.

Implementation of the invention

Definition

Reference will now be made in detail to certain embodiments, examples of which are illustrated in the appended structures and claims. Although the enumerated embodiments will be described, it is to be understood that they are not intended to limit the invention to these embodiments. On the contrary, the invention is intended to cover all alternatives, modifications and equivalents that may be included within the scope of the present invention, as defined in the claims. One skilled in the art would recognize many methods and materials similar or equivalent to the methods and materials described herein that could be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described herein. To the extent that one or more of the incorporated literature and similar materials differs from or conflicts with this application, including, without limitation, defined terms, use of terms, described techniques or the like, this application shall control.

The term “alkyl” means a straight or branched monovalent saturated hydrocarbon containing, unless otherwise stated, 1-20 carbon atoms. Numerical ranges in this specification are intended to include any number(s) within the specified range, as if the individual numbers were separately disclosed. For example, an alkyl group of 1-20 carbons would include C1, C2, ...C20, as well as C1-C20, C1-C15, C1-C10, C1-C6, C1-C4, etc. Examples of alkyls include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, and t-butyl.

The term "alkenyl" means a straight or branched monovalent hydrocarbon containing 2-20 hydrocarbon atoms (e.g., C2-C10) and one or more double bonds. Examples of alkenyls include, but are not limited to, ethenyl, propenyl, allyl, and 1,4-butadienyl.

The term "alkynyl" means a straight or branched monovalent hydrocarbon containing 2-20 hydrocarbon atoms (e.g., C2-C10) and one or more triple bonds. Examples of alkynyls include, but are not limited to, ethynyl, 1-propynyl, 1- and 2-butynyl, and 1-methyl-2-butynyl.

The term "aloxy" means an -O-alkyl radical in which the alkyl portion is as defined above. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, iso-butoxy, sec-butoxy, and tert-butoxy.

The term "acyloxy" means an -O-C(O)-R radical in which R may be H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl or heteroaryl.

The term "amino" means NH ₂ . The term "alkylamino" means an -N(R)-alkyl radical in which R may be H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl or heteroaryl.

The term "cycloalkyl" means a monovalent saturated hydrocarbon ring system having from 3 to 30 carbon atoms (e.g., C3-C6 or C3-C12). Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and adamantanyl.

The term "cycloalkenyl" means a monovalent non-aromatic hydrocarbon ring system having from 3 to 30 carbon atoms (e.g., C3-C6 or C3-C12) and one or more double bonds. Examples include cyclopentenyl, cyclohexenyl, and cycloheptenyl.

The term "heterocycloalkyl" means an aromatic 5-8 membered monocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ring system having one or more heteroatoms (such as O, N, S, or Se). Examples of heterocycloalkyl groups include, but are not limited to, piperazinyl, pyrrolidinyl, piperidinyl, dioxanyl, morpholinyl, and tetrahydrofuranyl.

The term "heterocycloalkenyl" means an aromatic 5- to 8-membered monocyclic, 8- to 12-membered bicyclic, or 11- to 14-membered tricyclic ring system having one or more heteroatoms (such as O, N, S, or Se) and one or more double bonds.

The term "aryl" means a monovalent 6-carbon monocyclic, 10-carbon bicyclic, or 14-carbon tricyclic aromatic ring system. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, and anthracenyl.

The term "aryloxyl" means -O-aryl. The term "arylamino" means -N(R)-aryl, wherein R can be H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, or heteroaryl. The term "heteroaryl" refers to an aromatic 5-8-membered monocyclic, 8-12-membered bicyclic, or 11-14-membered tricyclic ring system having one or more heteroatoms (such as O, N, S, or Se). Examples of heteroaryl groups include, but are not limited to, pyridyl, furyl, imidazolyl, benzimidazolyl, pyrimidinyl, thienyl, quinolinyl, indolyl, thiazolyl, pyrrolyl, isoquinolinyl, purinyl, oxazolyl, pyrazolyl, and carbazolyl. In all these terms, the "aryl" portion is as defined above.

The term "halogen" means F, Cl, Br or I. Preferably, it is F, Cl or Br.

The above-described alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, amino, alkylamino, arylamino, alkoxy, aryloxy, aryl and heteroaryl moieties may be substituted or unsubstituted moieties. Possible substituents of amino, alkylamino, arylamino, alkoxy, aryloxy, cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl and heteroaryl include, without limitation, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C20 cycloalkyl, C3-C20 cycloalkenyl, C1-C20 heterocycloalkyl, C1-C20 heterocycloalkenyl, C1-C10 alkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, amino, C1-C10 alkylamino, arylamino, hydroxy, halo, oxo (O=), thioxo (S=), thio, C1-C10 alkylthio, arylthio, C1-C10 alkylsulfonyl, arylsulfonyl, acylamino, aminoacyl, aminothioacyl, amidino, mercapto, amido, thioureido, thiocyanato, sulfonamido, guanidine, ureido, cyano, nitro, acyl, thioacyl, acyloxy, carbamido, carbamyl (-C(O)NH ₂ ), carboxyl (-COOH) and carboxylic ether. On the other hand, possible substitutes for alkyl, alkenyl or alkynyl include all of the above except C1-C10 alkyl. Cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl and heteroaryl can also be fused with each other.

Provided herein are compounds and pharmaceutical compositions thereof that are useful in the treatment or prevention of diseases, conditions and/or disorders modulated/mediated by (or associated with) type III receptor tyrosine kinases such as FLT3, PDGFR, c-KIT and/or CSF-1R kinases.

In some embodiments of the invention, compounds of formula I are provided:

Formula (1)

or a stereoisomer, tautomer, pharmaceutically acceptable salt, proteolysis-targeted chimera (PROTAC) linker thereof, in which

X is selected from the group consisting of CR ² and NR ³ ;

Y is selected from the group consisting of CR ² and NR ³ ;

G is selected from the group consisting of optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, alkynyl, and direct bond;

L is selected from the group consisting of ^-CHR4- , -( _CH2 ) _q- , ^-NR5- , -O-, -O( _CH2 ) _q- , -C(O)-, -C(O)( _CH2 ) _q- , -C(O)NH-, _-SO2- , and a direct bond, wherein each q is individually and independently an integer from 1 to 4;

Z is selected from the group consisting of hydrogen, C1-C4 alkyl, C1-C4 alkylamine, C1-C4 dialkylamine, optionally substituted four-, five- and six-membered cyclic amine, and optionally substituted heterocyclyl and heterocyclic spiro compound;;

R ¹ is selected from the group consisting of hydrogen, deuterium, halogen, hydroxyl, amino, cyano, trifluoromethyl, trifluoromethoxy, nitro, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino and C1-C6 dialkylamino;

R ² is selected from the group consisting of hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C5-C6 cycloalkenyl, C1-C6 alkylamino, C1-C6 dialkylamino, C3-C6 cycloalkylamino, C1-C6 alkoxy, C3-C6 cycloalkoxy, aryl, 3-6-membered heterocyclyl, and 5-6-membered heteroaryl, wherein the alkylamino, dialkylamino, alkoxy, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkylamino, cycloalkoxy, heterocyclyl, aryl and heteroaryl are optionally substituted with halogen, amino, hydroxyl, cyano, nitro, acrylic, acyloxy, C1-C4 alkyl, C1-C4 hydroxyl alkyl, C1-C4 alkoxy alkyl, C1-C4 acyloxy alkyl, C1-C4 alkoxy, C1-C4 dialkylamino, C3-C6 cycloalkyl, 3-6-membered heterocyclyl, aryl and 5-6-membered heteroaryl;

R ³ is selected from the group consisting of hydrogen, deuterium, C1-C6 alkyl, C1-C6 alkylcarbonyl, C2-C6 alkenylcarbonyl, C1-C6 alkoxycarbonyl, aminocarbonyl, C1-C6 alkylaminocarbonyl and C1-C6 dialkylaminocarbonyl;

R ⁴ is selected from the group consisting of hydrogen, deuterium, C1-C6 alkyl; and

R ⁵ is selected from the group consisting of hydrogen and C ₁ -C ₆ alkyl.

One skilled in the art should understand that in the above formula (I) all possible combinations or permutations of the various substituents are within the scope of the invention. These compounds can be prepared using readily available materials/reagents and known chemical reactions. Based on general knowledge in the art and the teaching of this disclosure, one skilled in the art should be able to prepare and use these compounds without undue experimentation.

The following reaction schemes, Scheme 1 through Scheme 6, provide typical procedures that can be used to prepare compounds of Formula (I). However, one skilled in the art will understand that these examples are provided for illustrative purposes only and that modifications or variations are possible without departing from the scope of the invention. The heterocyclic pyrazole compound synthesized according to embodiments of the invention can be purified by any known methods, such as flash column chromatography, high performance liquid chromatography, crystallization, or any other suitable methods.

Synthesis Path I

Scheme 1

Boric acid or boronate (1.2 equiv.) was added to a solution of 2-Bromo-5,6-dihydro-4H-cyclopenta[b]thiophene-4-one (1.0 equiv.) in 1,4-dioxane/water (5:1). K ₂ CO ₃ (2 equiv.) and Suzuki catalyst with a combination of Pd(II) or Pd(0) (5~8 mol %) (e.g. Pd(dppf)Cl ₂ , Pd(PPh ₃ ) ₄ , or Pd(OAc) ₂ , etc.) were added to the solution. The resulting mixture was heated under reflux. After completion of the reaction, water was added and the solution was extracted with ethyl acetate. The organic layers were dried over MgSO ₄ and evaporated in vacuo. The final compound was obtained after purification by chromatography.

Scheme 2

60% sodium hydride (NAH, 1.3 equiv) was added to a suspension of the active imidazole ester (1.1 equiv) and dihydrocyclopenta[b]thiophene-4-one derivative (1.0 equiv) in tetrahydrofuran (THF, 0.2 M) under nitrogen atmosphere. The reaction was then heated under reflux for 4 h. After completion of the reaction, the reaction was quenched with acetic acid (HOAc, 1.5 equiv) at 0 °C. The resulting yellow precipitate was then filtered and washed with water and cold methanol. The solid was then dried in vacuo to afford the desired 1,3-diketone compound.

Scheme 3

Hydrazine hydrate (3 equiv) was added to a suspension of the diketone (1 equiv) and acetic acid (HOAc, 5 equiv) in ethanol (EtOH, 0.2 M). The reaction was then heated under reflux for 6 h. After completion of the reaction, the resulting yellow precipitate was then filtered and washed with water and cold methanol. The solid was then dried in vacuo to afford the desired pyrazole compound.

Synthesis Path II

Scheme 4

60% sodium hydride (NAH, 1.3 equiv) was added to a suspension of the active imidazole ester (1.1 equiv) and dihydrocyclopenta[b]thiophene-4-one derivative (1.0 equiv) in tetrahydrofuran (THF, 0.2 M) under nitrogen atmosphere. The reaction was then heated under reflux for 4 h. After completion of the reaction, the reaction was quenched with acetic acid (AcOH, 1.5 equiv) at 0 °C. The resulting precipitate was then filtered and washed with water and cold methanol. The solid was then dried in vacuo to afford the desired 1,3-diketone compound.

Scheme 5

Hydrazine hydrate (3 equiv) was added to a suspension of the diketone (1 equiv) and acetic acid (HOAc, 5 equiv) in ethanol (EtOH, 0.2 M). The reaction was then heated under reflux for 6 h. After completion of the reaction, the resulting precipitate was then filtered and washed with water and cold methanol. The solid was then dried in vacuo to afford the desired pyrazole compound.

Scheme 6

Boric acid or boronate (1.2 equiv.) was added to a solution of pyrazole derivative (1.0 equiv.) in 1,4-dioxane/water (5:1). Potassium carbonate ( _{K2CO3 ,} 2 equiv.) and a conventional Suzuki catalyst with a combination of Pd(II) or Pd(0) (5~8 mol%) (e.g. Pd(dppf) _Cl2, Pd( _PPh3 ) ₄ , or Pd(OAc) ₂ , etc.) were added to the solution. The resulting mixture was heated under reflux. After completion of the reaction, water was added and the solution was extracted with ethyl acetate. The organic layers were dried over _MgSO4 and evaporated in vacuo. The final compound was obtained after purification by chromatography.

The above reaction schemes illustrate how heterocyclic pyrazole compounds can be prepared. One skilled in the art will recognize that the envisioned reactions and the reagents used in these reactions are known in the art. Therefore, based on the above teachings and the general knowledge in the art, heterocyclic pyrazole compounds with various substituents as defined herein can be prepared by one skilled in the art without inventive efforts.

The compounds of Formula I are novel, potent inhibitors of type III receptor tyrosine kinases, such as FLT3, PDGFR, c-KIT and/or CSF-1R kinases, and may be useful in the prevention and treatment of disorders resulting from the action of these kinases.

Compounds of Formula I may have therapeutic value in the treatment of diseases or disorders selected from fibrosis, bone-related diseases, cancer, autoimmune disorders, inflammatory diseases, and cardiovascular diseases.

According to some embodiments of the invention, the compounds of Formula I are useful for treating fibrosing diseases. Examples of fibrosis include idiopathic pulmonary fibrosis (IPF), nephrogenic gene systemic fibrosis (NSF), liver cirrhosis, diabetes-induced nephropathy, cardiac fibrosis (e.g., endomyocardial fibrosis), mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis, Crohn's disease, keloid formation, scleroderma, and systemic sclerosis. Additional examples of fibrotic diseases include focal segmental glomerulosclerosis (FSGS), systemic sclerosis-interstitial lung disease (SSC-ILD), primary biliary cirrhosis, ethanol cirrhosis, interstitial fibrosis and tubular atrophy (CAD), proliferative vitreoretinopathy, and scarring (hypertrophic and keloid).

According to some embodiments of the invention, the compounds of Formula I are useful for treating bone-related diseases, including metastatic bone disease, treatment-related bone loss, osteoporosis, rheumatoid arthritis, ankylosing spondylitis, Paget's disease, and periodontal disease. Osteoporosis can be associated with (1) menopause in women, (2) aging in men or women, (3) suboptimal bone growth during childhood and adolescence resulting in failure to achieve peak bone mass, and/or (4) bone loss secondary to other diseases, eating disorders, medications, and/or therapeutic treatments (e.g., glucocorticoid therapy, aromatase inhibition therapy, or antiandrogen therapy).

Other osteolytic diseases that can be treated in accordance with embodiments of the present invention are more localized. A specific example is tumor-related osteolysis. In this condition, bone cancer or bone metastases cause localized osteolysis, which causes pain, weakness, or bone fractures. Such localized osteolysis also allows tumors to grow in size, creating more space for them in the bone and releasing growth factors from the bone matrix. Cancers that are now known to cause tumor-related osteolysis include hematological malignancies (e.g., myeloma and lymphoma) and solid tumors (e.g., breast, prostate, lung, kidney, and thyroid), all of which cancers are treated by the present invention.

According to some embodiments of the invention, the compounds of Formula I are useful for treating cancer and proliferative disorders. Examples include multiple myeloma, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), prostate cancer, breast cancer, ovarian cancer, melanoma, glioblastoma multiforme, giant cell tumor of bone (also known as osteoclastoma), giant cell tumor of the tendon sheath (also known as tenosynovial giant cell tumor or TGCT), metastasis of tumors to other tissues, other chronic myeloproliferative diseases such as myelofibrosis and pigmented villonodular synovitis (PVNS).

According to some embodiments of the invention, the compounds of Formula I are useful for treating autoimmune disorders and inflammatory diseases, including, but not limited to, rheumatoid arthritis, osteoarthritis, psoriatic arthritis, ankylosing spondylitis, adult-onset Still's disease, glomerulonephritis, osteoporosis, Sjogren's syndrome, inflammatory bowel disease, ulcerative colitis, Crohn's disease, Langerhans cell histiocytosis, hemophagocytic syndrome, multicentric reticulohistiocytosis, and Paget's disease. Additional examples of autoimmune diseases and disorders include primary sclerosing cholangitis and transplant rejection (including liver, kidney, and heart/lung transplants).

According to some embodiments of the invention, the compounds of Formula I are useful for treating cardiovascular diseases. Examples of cardiovascular diseases include atherosclerosis, peripheral vascular disease, coronary artery disease, ischemia/reperfusion, hypertension, restenosis, pulmonary arterial hypertension, and arterial inflammation. Further examples of cardiovascular diseases include acute respiratory distress syndrome (ARDS), arteriovenous (AV) fistula patency, and veno-occlusive disease (post-HSC/BMT).

The heterocyclic pyrazole compounds described herein may contain a non-aromatic double bond and one or more asymmetric centers, such as in substituents attached to the main aromatic rings. Accordingly, these compounds may occur as racemates and racemic mixtures, single enantiomers, individual diastereomers, diastereomeric mixtures, and cis or trans isomeric forms. All such isomeric forms are within the scope of the invention. The heterocyclic pyrazole compounds of the invention may have acidic or basic functional groups (e.g., on substituent groups) that can form salts, in particular pharmaceutically acceptable salts. The formation of such salts is routine practice in the pharmaceutical industry. Examples of salts that can be used with the heterocyclic pyrazole compounds of the invention include chloride, sulfate, mesylate, besylate, tosylate, formate, acetate, malate, succinate, etc. for basic functional groups. Such heterocyclic pyrazole salts are within the scope of the invention. Likewise, acidic or basic groups can be functionalized, for example, into esters. Such functionalized derivatives will be hydrolyzed in vivo. Therefore, such derivatives can function as prodrugs of the heterocyclic pyrazole compounds of the invention. The formation of prodrugs involves only routine skill, and one skilled in the art should know how to prepare and use such prodrugs without undue experimentation.

Also within the scope of the present invention are (1) a pharmaceutical composition that comprises an effective amount of at least one of the heterocyclic pyrazole compounds of the invention and a pharmaceutically acceptable carrier, (2) a method of treating a disease associated with a protein kinase (e.g., cancer) by administering to a subject in need of such treatment an effective amount of such a heterocyclic pyrazole compound, and (3) a method of reducing the activity of at least one protein kinase by contacting at least one protein kinase with at least one of the heterocyclic pyrazole compounds of the present invention.

As used herein, the term "protein kinase-associated disease/disorder" or "protein kinase-related disease/disorder" or "protein kinase-modulated disease/disorder" refers to a disease or condition that is characterized by abnormal protein kinase (PK) activity, or to a disease or condition that can be treated by altering the activity of at least one PK. Abnormal PK activity may result from increased PK expression or from the presence of PK expression that does not occur under normal conditions. The PC-related diseases/disorders described herein include, but are not limited to, cancer, diabetes, hyperproliferation disorder, hyperproliferative disorders of the kidney, kidney disease, von Hippel-Lindau disease, restenosis, fibrosis, psoriasis, osteoarthritis, rheumatoid arthritis, inflammatory disease, immunological disorders such as autoimmune diseases (e.g., AIDS, lupus, etc.), cardiovascular disorders (e.g., atherosclerosis), and proliferative disorders of blood vessels such as abnormal vasculogenesis.

The term "treatment" means administering a heterocyclic pyrazole compound of the invention to a subject having a protein kinase-related disease/disorder, or having symptoms thereof, or a predisposition thereto, for the purpose of treating, healing, alleviating, alleviating, modifying, eliminating, ameliorating, arresting, affecting, or reducing the risk of the disorder, the symptoms of the disorder, or the predisposition thereto. For example, treating cancer means treating that results in inhibiting the growth of cancer or the growth of cancer cells, causing regression of cancer growth (i.e., it reduces the size of detectable cancer), or causing the cancer to disappear.

The term "effective amount" means the amount/volume of active substance required to produce the intended therapeutic effect in a subject. Effective amounts may vary, as recognized by those skilled in the art, depending on the routes of administration, the use of excipients, and the possibility of use with other substances. Determination of an effective amount requires only routine skill, and one skilled in the art should be able to determine such effective amounts for the intended use without undue experimentation. The subject to be treated must be a mammal. The term "mammal" means a human or non-human mammal, such as a dog, cat, pig, cow, sheep, donkey, horse, rat, or mouse.

To carry out the method of the present invention, any of the pharmaceutical compositions described above can be administered orally, parenterally, via inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implanted reservoir. The term "parenteral" as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intra-arterial, intra-synovial, intrasternal, interstitial, intralesional, and intracranial injection or infusion methods. According to some embodiments of the invention, the heterocyclic pyrazole compound of the present invention can be administered intravenously, suitable carriers can include, but are not limited to, physiological saline or sodium phosphate buffered saline (PBS) and solutions containing thickening and solubilizing components, such as glucose, polyethylene glycol, and polypropylene glycol, and mixtures thereof.

A sterile injectable composition, for example a sterile injectable aqueous or oleaginous suspension, can be prepared according to techniques known in the art using suitable dispersing or wetting agents (for example, TWEEN 80) and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable carriers and solvents that can be used are mannitol, water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils (for example, synthetic mono- or diglycerides) are commonly used as a solvent or suspending medium. Fatty acids such as oleic acid and its glyceride derivatives are suitable for the preparation of injectables, as are natural pharmaceutically acceptable oils such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain an alcoholic diluent or a long-chain dispersant, or carboxymethylcellulose or similar dispersants. For the purposes of preparing the dosage form, other commonly used surfactants may also be used, such as Tweens or Spans or other similar emulsifying agents or bioavailability enhancers that are commonly used in the manufacture of pharmaceutically acceptable solid, liquid or other dosage forms.

The composition for oral administration may be any orally acceptable dosage form, including, but not limited to, capsules, tablets, emulsions and aqueous suspensions, dispersions and solutions. In the case of tablets for oral use, carriers that are commonly used include lactose and corn starch. Lubricants, such as magnesium stearate, are also commonly added. For oral administration in capsule form, diluents used include lactose and dried corn starch. In the oral administration of aqueous suspensions or emulsions, the active ingredient can be suspended or dissolved in an oil phase in combination with emulsifying or suspending agents. If desired, certain sweetening, flavoring or coloring agents can be added. The nasal aerosol or inhalation composition can be prepared according to methods well known in the pharmaceutical manufacturing art. The composition containing the heterocyclic pyrazole compound can also be administered in the form of suppositories for rectal administration.

The carrier in the pharmaceutical composition must be "acceptable" in the sense of being compatible with the active ingredient of the dosage form (and preferably capable of stabilizing it) and not causing harm to the subject to be treated. One or more solubilizing components (e.g., cyclodextrin) that form more soluble complexes with the active heterocyclic pyrazole components can be used as pharmaceutical carriers for delivery of the active components. Examples of other carriers include colloidal silicon dioxide, magnesium stearate, and sodium lauryl sulfate.

The above reaction schemes illustrate how heterocyclic pyrazole compounds can be prepared. One skilled in the art will recognize that the envisioned reactions and the reagents used in these reactions are known in the art. Therefore, based on the above teachings and the general knowledge in the art, heterocyclic pyrazole compounds with various substituents as defined herein can be prepared by one skilled in the art without inventive efforts.

It is believed that the above description has adequately enabled the present invention to be carried out without further elaboration. Accordingly, the following examples are to be construed as merely illustrative and in no way limiting of the remainder of the disclosure.

EXAMPLE 1: Synthesis of compounds of formula (I)

The exemplified heterocyclic pyrazole compounds are listed in Table 1. Their ¹ H NMR and mass data are presented in Table 2.

Table 1. Heterocyclic pyrazole compounds

Example Example Structure Name A 3-(1H-indol-6-yl)-6-(6-methoxypyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole B 3-(1H-indol-5-yl)-6-(6-methoxypyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole C 3-(1H-benzo[d]imidazol-6-yl)-6-(6-methoxypyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole D 3-(indolin-6-yl)-6-(6-methoxypyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole E 3-(indolin-5-yl)-6-(6-methoxypyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole F 3-(benzofuran-6-yl)-6-(6-methoxypyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole G 3-(benzofuran-5-yl)-6-(6-methoxypyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole H 5-(3-(4-methoxyphenyl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-amine I 3-(4-methoxyphenyl)-6-(6-methoxypyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole J N-(1H-indazol-5-yl)-6-(6-methoxypyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-3-amine K 3-(4-methoxyphenyl)-6-(6-methylpyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole

Working example Compound No. Structure Name 1 3-(1H-indazol-5-yl)-6-(6-methyloxypyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 2 3-(1H-indazol-6-yl)-6-(6-methoxypyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 3 5-(3-(1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-amine 4 6-(6-methoxypyridin-3-yl)-3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 5 N,N-dimethyl-5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-amine 6 4-(4-(3-(1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)benzyl) morpholine 7 (2S,6R)-4-(4-(3-(1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol- 6-yl)benzyl)-2,6-dimethylmorpholine 8 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)morpholine 9 (2S,6R)-2,6-dimethyl-4-((5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-yl)methyl)morpholine 10 4-((5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-yl)methyl)morpholine

11 3-(1-methyl-1H-indazol-6-yl)-6-(6-methylpyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 12 (2S,6R)-2,6-dimethyl-4-((3-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4.5 ]cyclopenta[1,2-c]pyrazol-6-yl)benzyl)morpholine 13 4-((3-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6 -yl)benzyl)morpholine 14 3-(1-methyl-1H-indazol-6-yl)-6-(3-((4-methylpiperazin-1-yl)methyl)phenyl)-1,4-dihydrothieno[2',3':4, 5]cyclopenta[1,2-c]pyrazole 15 (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(4-methylpiperazin-1-yl)methanone 16 (2R,6S)-4-((5-(3-(1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-yl)methyl)-2,6-dimethylmorpholine 17 3-(1-methyl-1H-indazol-6-yl)-6-(2-methylpyridin-4-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 18 4-((5-(3-(1H-indazol-6-yl)-1,4-dihydrolthieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridine -2-yl)methyl)morpholine 19 ((2S,6R)-2,6-dimethylmorpholino)(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)methanone 20 4-(1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole -6-yl)phenyl)ethyl)morpholine 21 (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(morpholino)methanone 22 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenethyl)morpholine 23 (5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopental[1,2-c]pyrazol-6-yl)pyridin-2-yl)(morpholino)methanone 24 4-(2-methoxy-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazol-6-yl)benzyl)morpholine 25 4-(2-fluoro-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazol-6-yl)benzyl)morpholine 26 4-(2-methyl-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazol-6-yl)benzyl)morpholine 27 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)-2-(trifluoromethyl)benzyl)morpholine 28 4-(1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)propyl)morpholine 29 4-(3-methoxy-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazol-6-yl)benzyl)morpholine 30 3-(1H-indazol-6-yl)-6-(6-methylpyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 31 (5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-3-yl)(morpholino)methanone 32 4-(3-fluoro-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazol-6-yl)benzyl)morpholine 33 3-(1-methyl-1H-indazol-6-yl)-6-(6-methylpyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 34 (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(piperidin-1-yl)methanone 35 4-(4-(3-(1-isobutyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)morpholine 36 3-(1H-indazol-5-yl)-6-(6-methylpyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 37 (3-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(morpholino)methanone 38 (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(pyrrolidin-1-yl)methanone 39 3-(1-methyl-1H-indazol-6-yl)-6-(4-(thiomorpholinomethyl)phenyl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c ]pyrazole 40 5-(3-(1H-indazol-5-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-amine 41 (5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyrimidin-2-yl)(morpholino)methanone 42 4-(4-(3-(1-isopropyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)morpholine 43 (2-methoxy-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(morpholino)methanone 44 (2-fluoro-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(morpholino)methanone 45 5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyrimidin-2-amine 46 (1-methyl-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)-1H-pyrrol-2-yl)(morpholino)methanone 47 4-((4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6 -yl)phenyl)sulfonyl)morpholine 48 (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)thiophen-2-yl)(morpholino)methanone 49 N,N-dimethyl-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole -6-yl)banzamide 50 N,N-dimethyl-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)benzenesulfonamide 51 4-((5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)thiophen-2-yl)methyl)morpholine 52 4-((4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)thiophen-2-yl)methyl)morpholine 53 4-(2,6-difluoro-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2- c]pyrazol-6-yl)benzyl)morpholine 54 (2S,6R)-2,6-dimethyl-4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5] cyclopental[1,2-c]pyrazol-6-yl)benzyl)morpholine 55 4-((6-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-3-yl)methyl)morpholine 56 (2-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-4-yl)(morpholino)methanone 57 (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)-3,6-dihydropyridin-1(2H)-yl)(morpholino)methanone 58 tert-butyl 4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)piperidine-1-carboxylate 59 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)morpholine-3-one 60 (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)thiazol-2-yl)(morpholino)methanone 61 1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)piperidine-2-one 62 3-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)oxazolidine-2-one 63 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)piperazine-2-one 64 1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)pyrrolidine-2-one 65 1,3-dimethyl-4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2- c]pyrazol-6-yl)benzyl)piperazine-2-one 66 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenethyl)morpholine-3-one 67 6-(4-((3,3-difluoropyrrolidin-1-yl)methyl)phenyl)-3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 68 4-(4-(3-(3-bromo-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)morpholine 69 4-(2-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole -6-yl)phenoxy)ethyl)morpholine 70 4-(2-(3-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole -6-yl)phenoxy)ethyl)morpholine 71 4-(3-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)prop-2-yn-1-yl)morpholine 72 N-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)tetrahydro-2H-pyran-4-amine 73 N-methyl-N-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)tetrahydro-2H-pyran-4-amine 74 (R)-3-methyl-4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1, 2-c]pyrazol-6-yl)benzyl)morpholine 75 (S)-3-methyl-4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1, 2-c]pyrazol-6-yl)benzyl)morpholine 76 3-(1-methyl-1H-indazol-6-yl)-6-(4-((tetrahydro-4H-puran-4-ylidene)methyl)phenyl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 77 3-(1-methyl-1H-indazol-6-yl)-6-(4-((tetrahydro-2H-puran-4-yl)methyl)phenyl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 78 4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)morpholine 79 4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)morpholine-3-one 80 4-(1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole -6-yl)benzyl)piperidin-4-yl)morpholine 81 4-(4-(3-(3-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)morpholine 82 4-(5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)pyridin-2-yl)morpholine 83 3-(1H-indazol-6-yl)-6-(4-((4-methylpiperazin-1-yl)methyl)phenyl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 84 3-(1-methyl-1H-indazol-6-yl)-6-(4-((4-methylpiperazin-1-yl)methyl)phenyl)-1,4-dihydrothieno[2',3':4, 5]cyclopenta[1,2-c]pyrazole 85 3-(1-methyl-1H-indazol-6-yl)-6-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 86 6-(2-chloropyridin-4-yl)-3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 87 3-(1-methyl-1H-indazol-6-yl)-6-(2-(4-methylpiperazin-1-yl)pyridin-4-yl)-1,4-dihydrothieno[2',3':4,5]cyclopental[1,2-c]pyrazole 88 4-(2,3-difluoro-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2- c]pyrazol-6-yl)benzyl)morpholine 89 4-(4-(3-(1-methyl-1H-benzo[d][1,2,3]triazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta [1,2-c]pyrazol-6-yl)benzyl)morpholine 90 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)-3-(trifluoromethoxy)benzyl)morpholine 91 4-(3-methyl-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazol-6-yl)benzyl)morpholine 92 5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-amine 93 N,N-dimethyl-1-(5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-yl)methanamine 94 N,N-dimethyl-1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)methanamine 95 4-((6-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-3-yl)methyl)morpholine 96 3-(1-methyl-1H-indazol-6-yl)-6-(piperidin-4-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazole 97 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)-1,4-oxazepane 98 1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)piperidin-1-yl)-2-morpholinoethane-1-one 99 3-(1-methyl-1H-indazol-6-yl)-6-(4-(pyrrolidin-1-ylmethyl)phenyl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 100 3-(1-methyl-1H-indazol-6-yl)-6-(4-(piperidin-1-ylmethyl)phenyl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 101 1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)piperazine-2-one 102 6-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)-2-oxa-6-azaspiro[3.3]heptane 103 6-(4-(azetidin-1-ylmethyl)phenyl)-3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1 ,2-c]pyrazole 104 6-(4-((3,3-difluoroazetidin-1-yl)methyl)phenyl)-3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole 105 6-(4-((3-methoxyazetidin-1-yl)methyl)phenyl)-3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4, 5]cyclopenta[1,2-c]pyrazole

Table 2. ¹ H NMR and mass data

Example Example NMR Mass (mass/charge) A ^1H NMR (600 MHz, DMSO-d6, ppm) δ 12.92 (s, 1H), 11.29 (s, 1H), 8.51 (d, J =2.3 Hz, 1H), 8.01 (dd, J =8.6, 2.4 Hz, 1H), 7.76 (s, 1H), 7.67 (s, 1H), 7.60 (d, J =8.2 Hz, 1H), 7.41-7.36 (m, 2H), 6.86 (d, J =8.6 Hz , 1H), 6.42 (s, 1H), 5.56 (s, 2H), 3.93 (s, 2H), 3.84 (s, 3H). 385.0 (M+H) ⁺ B ^1H NMR (500 MHz, DMSO-d6, ppm) δ 12.86 (s, 1H), 11.24 (s, 1H), 8.56 (s, 1H), 8.08-8.04 (m, 1H), 7.98 (s, 1H), 7.70 (s, 1H), 7.56-7.48 (m, 2H), 7.42 (s, 1H), 6.95-6.89 (m, 1H), 6.51 (s, 1H), 3.98 (s, 2H), 3.91 (s, 3H). 385.3 (M+H) ⁺ C ^1H NMR (600 MHz, DMSO-d6, ppm) δ 8.57-8.53 (m, 2H), 8.07-8.03 (m, 2H), 7.76 (d, J = 8.4 Hz, 1H), 7.72 (s, 1H), 7.70 (s, 1H), 6.92 (dd, J =8.6, 0.6 Hz, 1H), 4.01 (s, 2H), 3.90 (s, 3H). 386.5 (M+H) ⁺ D ¹ H NMR (500 MHz, DMSO-d6, ppm) δ 13.82 (s, 1H), 8.54 (d, J =2.5 Hz, 1H), 8.04 (dd, J =8.6, 2.5 Hz, 1H), 7.68 (s , 1H), 7.12 (d, J =7.5 Hz, 1H), 6.97-6.93 (m, 1H), 6.92-6.87 (m, 2H), 5.67 (s , 1H), 3.89 (s , 3H), 3.88 (s , 2H), 3.49-3.42 (m, 2H), 2.97-2.91 (m, 2H). 386.70 (M+H) ⁺ E ^1H NMR (600 MHz, DMSO-d6, ppm) δ 12.63 (s, 1H), 8.54 (d, J =2.5 Hz, 1H), 8.03 (dd, J =8 ,6, 2.5 Hz, 1H), 7.67 (s, 1H), 7.44 (s, 1H), 7.34 (d, J =8.0 Hz, 1H), 6.90 (d, J =8.6 Hz, 1H), 6.57 (d, J =8.0 Hz, 1H), 5, 77 (s, 1H), 5.67 (s, 1H), 3.89 (s, 3H), 3.87 (s, 2H), 3.50-3.45 (m, 2H), 3.00-2.96 (m, 2H). 387.22 (M+H) ⁺ F ¹ H NMR (600 MHz, DMSO-d6, ppm): δ 13.06 (s, 1H), 8.57-8.54 (m , 1H), 8.08-8.03 (m, 2H), 8.00 (s , 1H), 7.78 (d, J = 8.1 Hz, 1H), 7.72-7.66 (m, 2H), 7.03-7.00 (m, 1H), 6.93-6.89 (m, 1H), 4.02 (s, 2H), 3.90 (s , 3H) ppm. 386.09 (M+H) ⁺ G ^1H NMR (600 MHz, DMSO-d6, ppm): δ 13.02 (s, 1H), 8.55 (d, J =2.5 Hz, 1H), 8.08-8.03 ( m, 3H), δ 7.77-7.67 (m, 3H), 7.05 (d, J = 2.0 Hz, 1H), 6.90 (d, J =8.5 Hz, 1H), 3.99 (s, 2H), 3.90 (s, 3H) ppm. 386.03 (M+H) ⁺ H ^1H NMR (500 MHz, DMSO-d6, ppm) δ 8.49 (s, 1H), 8.00 (s, 1H), 7.93 (d, 1H), 7.82 (s, 1H ), 7.71 (d, 2H), 7.07 (d, 2H), 4.03 (s, 2H), 3.81(s,3H). 397.0 (M+H) ⁺ I ^1H NMR (500 MHz, DMSO-d6, ppm) δ 8.54 (d, 1H), 8.03 (d, 1H), 7.72-7.68 (m, 3H), 7.07 (d, 2H), 6.90 (d, H), 3.93 (s, 2H), 3.89 (s, 3H), 3.80(s, 3H). 376.0 (M+H) ⁺ . J ^1H NMR (500 MHz, DMSO-d6, ppm): δ 12.79 (sbr, 1H), 11.98 (sbr, 1H), 8.52 (d, J =2.5 Hz, 1H), 8.30-8.13 (m, 1H), 8.02 (dd, J =2.5, 8.5 Hz, 1H), 7.89 (s, 1H), 7.63 (s br, 1H), 7.42-7.41 (m, 1H), 7.15-7.09 (m, 2H), 6.90 (d, J =8.5 Hz, 1H), 3.89 (s, 2H), 3.57 (s, 3H). 401.07 (M+H) ⁺ . K ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 12.91 (s, 1H), 8.81 (d, J =2.4 Hz, 1H), 7.99 (dd, J =2.4, 8.4 Hz, 1H), 7.80 (s, 1H), 7.70 (d, J =8.4 Hz, 2H), 7.31 (d, J =7.8 Hz, 1H), 7.06 (d, J =8.4 Hz, 2H), 3.94 (s, 2H), 3.80 (s, 3H). 360.42 (M+H) ⁺

Working example Compound No. NMR Mass (mass/charge) 1 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.23 (s, 1H), 13.02 (s, 1H), 8.56 (d, J =2.3 Hz, 1H), 8.20-8.15 (m, 2H), 8.06 (dd, J =8.6, 2.3 Hz, 1H), 7.80 (d, J =7.3 Hz, 1H), 7.75-7.64 (m, 2H), 6.92 (d, J =8.6 Hz, 1H), 4.01 (s, 2H), 3.91 (s, 3H). 385.6 (M) ⁺ 2 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.28 (s, 1H), 13.16 (s, 1H), 8.57 (d, J = 1.9 Hz, 1H), 8.11 (t, J =8.9 Hz, 1H), 8.06 (dd, J =8.9, 2.5 Hz, 1H), 7.90 (dd, J =8.5, 5.8 Hz, 1H), 7.72 (s, 1H), 7.59 (dd, J =37.4, 8.1 Hz, 1H ), 6.92 (d, J =8.6 Hz, 1H), 5.79 (s, 1H), 4.07 (s, 1H), 4.03 (s, 1H), 3.91 (s, 3H). 386.2 (M+H) ⁺ 3 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 8.29 (d, J =2.5 Hz, 1H), 8.12 (s, 1H), 7.93 (s, 1H), 7.89 (d, J =8.4 Hz, 1H), 7.73 (dd, J =8.6, 2.5 Hz, 1H), 7.55 (dd, J =8.4, 1.1 Hz, 1H), 7.49 (s, 1H), 6.52 (d, J =8.6 Hz, 1H), 6, 19 (s, 2H), 3.99 (s, 2H). 371.0 (M+H) ⁺ 4 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.14 (s, 1H), 8.56 (d, J =2.5 Hz, 1H), 8.09 (s, 1H), 8.06 (dd, J =8.6, 2.5 Hz, 1H), 8.01 (s, 1H), 7.89 (d, J =8.2 Hz, 1H), 7.64 (s, 1H), 7.56 (d, J =8.2 Hz, 1H), 6.92 (d, J =8.6 Hz , 1H), 4.11 (s, 3H), 4.08 (s, 2H), 3.91 (s, 3H). 399.9 (M) ⁺ 5 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.10 (s, 1H), 8.47 (d, J =2.3 Hz, 1H), 8.09 (s, 1H), 8.00 (s, 1H), 7.88 (d, J =8.3 Hz, 1H), 7.83 (d, J =8.9, 2.5 Hz, 1H), 7.58-7.54 (m, 2H), 6.71 (d, J =8.9 Hz, 1H), 4.11 (s, 3H) , 4.04 (s, 2H), 3.07 (m, 6H). 413.1 (M+H) ⁺ 6 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.30 (s, 1H), 13.17 (s, 1H), 8.12 (s, 1H), 7.93 (s, 1H), 7.90 (d, J =8.4 Hz, 1H), 7.73 (s, 1H), 7.70 (d, J =8.0 Hz, 2H), 7.55 (d, J =8.4 Hz, 1H), 7.37 (d, J =8.0 Hz, 2H), 4.03 (s, 2H), 3.59 (t, J =4.1 Hz, 4H), 3.49 (s, 2H), 2.38 (s, 4H). 453.9 (M) ⁺ 7 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.31 (s, 1H), 13.17 (s, 1H), 8.12 (s, 1H), 7.93 (s, 1H ), 7.90 (d, J =8.1 Hz, 1H), 7.73 (s, 1H), 7.70 (d, J =7.9 Hz, 2H), 7.55 (d, J =8.1 Hz, 1H), 7.36 (d, J =7.9 Hz, 2H), 4.03 (s , 2H), 3.60 - 3.53 (m, 2H), 3.46 (s, 1H), 3.17 (d, J =5.2 Hz, 1H), 2.69 (d, J =10.7 Hz, 2H), 1.66 (t, J =10.7 Hz, 2H), 1.04 (s, 3H), 1.03 (s, 3H). 482.0 (M+H) ⁺ 8 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.14 (s, 1H), 8.10 (s, 1H), 8.01 (s, 1H), 7.89 (d, J = 8.3 Hz, 1H), 7.73 (s, 1H), 7.70 (d, J = 7.9 Hz, 2H), 7.57 (d, J = 8.3 Hz, 1H), 7.37 (d, J = 7.9 Hz, 2H), 4.11 (s, 3H), 4.08 (s, 2H), 3.59 (t, J = 4.1 Hz, 4H), 3.49 (s, 2H), 2.38 (s, 4H). 467.8 (M) ⁺ 9 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 8.90 (d, J = 1.9 Hz, 1H), 8.10 (dd, J = 7.6, 1.6 Hz, 2H), 8.01 (s, 1H), 7.91 - 7.85 (m, 2H), 7.58 (d, J = 8.6 Hz, 1H), 7.50 (d, J = 8.2 Hz, 1H), 4.12 (s, 3H), 3.61 (s, 2H), 2.73 (d, J = 10.2 Hz, 2H), 2.65-2.60 (m, 1H), 2.41-2.37 (m, 1H), 1.78-1.74 (m, 2H), 1.04 (d, J =6.3 Hz, 6H). 496.8 (M) ⁺ 10 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.16 (s, 1H), 8.89 (s, 1H), 8.09 (d, J =2.4 Hz, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.88 (s, 1H), 7.87 (s, 1H), 7.55 (d, J =8.4 Hz, 1H), 7.50 (d, J =7.8 Hz, 1H), 4.10 (s, 5H), 3.61 -3.60 (m, 6H), 2.43 (s, br, 4H). 468.95 (M) ⁺ 11 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.15 (s, 1H), 8.83 (d, J =2.4 Hz, 1H), 8.08 (s, 1H), 8.01-7.99 (m, 2H), 7.87 (d, J =8.4 Hz, 1H), 7.83 (s, 1H), 7.55 (dd, J =0.6, 8.4 Hz, 1H), 7.32 (d, J =8.4 Hz, 1H), 4.10 (s, 3H), 4.09 (s, 2H), and the methyl signal on pyridine is hidden in the DMSO signal. 384.36 (M+H) ⁺ 12 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.12 (s, 1H), 8.08 (s, 1H), 8.03 (s, 1H), 8.00 (s, 1H ), 7.88 (d, J =8.3 Hz, 1H), 7.73 (s, 1H), 7.63 (s, 1H), 7.57 (d, J =5.3 Hz, 1H), 7.39 (d, J =8.3 Hz, 1H), 7.26 (d, J =7.5 Hz , 1H), 4.11 (s, 3H), 4.08 (s, 2H), 3.59 (d, J =1.5 Hz, 1H), 3.50 (s, 2H), 3.16 (d, J =5.2 Hz, 1H), 2.71 (d, J =10.3 Hz, 2H), 1.68 (t, J =10.8 Hz, 2H), 1.03 (d, J =6 ,3 Hz, 6H). 496.23 (M+H) ⁺ 13 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.13 (s, 1H), 8.09 (s, 1H), 8.00 (s, 1H), 7.88 (d, J =8.3 Hz, 1H), 7.74 (s, 1H), 7.66-7.62 (m, 2H), 7.59-7.55 (m, 1H), 7.42-7.38 (m, 1H), 7.27 (d, J =7.6 Hz, 1H), 4.11 (s, 3H) , 4.08 (s, 2H), 3.58 (t, J =4.5, 4.5 Hz, 4H), 3.53 (s, 2H), 2.42-2.37 (m, 4H). 468.37 (M+H) ⁺ 14 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.87 (d, J =8.4 Hz, 1H), 7.72 (s, 1H), 7.63-7.62 (m, 2H), 7.55 (d, J =8.4 Hz, 1H), 7.38 (t, J =8.4 Hz, 1H), 7.24 (d, J =7.2 Hz, 1H), 4, 10 (s, 3H), 4.07 (s, 2H), 3.51 (s, 2H), 3.31 (s br, 8H), 2.16 (s, 3H). 481.40 (M+H) ⁺ 15 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.15 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.88 (d, J =8.4 Hz, 1H), 7.85 (s, 1H), 7.80 (d, J =8.1 Hz, 2H), 7.56 (d, J =8.4 Hz, 1H), 7.45 (d, J =8.1 Hz, 2H), 4.11 (s, 3H), 4.10 (s, 2H), 3.62 (s, 2H), 3.39 (s, 2H), 2.33 (d, J =19.2 Hz, 4H), 2.20 (s, 3H). 495.41 (M+H) ⁺ 16 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.29 (s, 1H), 13.19 (s, 1H), 8.90 (d, J =2.2 Hz, 1H), 8.12 (s, 1H), 8.10 (dd, J =8.1, 2.2 Hz, 1H), 7.94 (s, 1H), 7.90 (d, J =8.3 Hz, 1H), 7.87 (s, 1H), 7.56 (d, J =7.5 Hz, 1H), 7.49 (d, J =8.1 Hz, 1H), 4.06 (s, 2H), 3.62-3.59 (m, 5H), 2.73 (d, J =10.6 Hz, 2H), 1.76 (t, J =10.6 Hz, 2H), 1.05 (s, 3H), 1.04 (s, 3H). 483.36(M+H) ⁺ 17 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.17 (s, 1H), 8.44 (d, J =5.4 Hz, 1H), 8.08 (s, 1H), 8.04 (s, 1H), 8.00 (s, 1H), 7.87 (d, J =8.4 Hz, 1H), 7.59 (s, 1H), 7.55 (d, J =8.4 Hz, 1H), 7.53 (d, J =4.8 Hz, 1H), 4.12 (s, 2H), 4.10 (s, 3H), 3.30 (s, 3H). 384.33 (M+H) ⁺ 18 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.27 (s, 1H), 13.17 (s, 1H), 8.89 (s, 1H), 8.09 (s, 1H ), 8.08 (dd, J =4.8, 7.2 Hz, 1H), 7.92 (s, 1H), 7.88 (d, J =8.4 Hz, 1H), 7.86 (s, 1H), 7.53 (d, J =8.4 Hz, 1H), 7.50 (d, J =8 .4 Hz, 1H), 4.05 (s, 2H), 3.61-3.59 (m, 6H), 2.44-2.43 (m, 4H). 455.23 (M+H) ⁺ 19 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.16 (s, 1H), 8.10 (s, 1H), 8.02 (s, 1H), 7.89 (d, J =8.4 Hz, 1H), 7.87 (s, 1H), 7.82 (d, J =8.2 Hz, 2H), 7.57 (d, J =8.3 Hz, 1H), 7.48 (d, J =8.2 Hz, 2H), 4.38 (s, 1H), 4.12 (s, 3H), 4.11 (s, 2H), 3.57 (s, 4H), 2.86 (s, 1H), 1.15 (s, 3H). 1.02(s, 3H). 509.97 (M) ⁺ 20 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.87 (d, J =8.4 Hz, 1H), 7.71 (s, 1H), 7.68 (d, J =7.8 Hz, 2H), 7.55 (d, J =8.4 Hz, 1H), 7.35 (d, J =7.8 Hz, 2H), 4.10 (s, 3H), 4.06 (s, 2H), 3.56-3.54 (m, 4H), 3.36 (q, J =6.6 Hz, 1H), 2.40 (s br, 2H), 2.30-2.27 (m, 2H), 1.29 (d, J = 6.6 Hz, 3H). 482.25 (M+H) ⁺ 21 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.15 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.91-7.83 (m, 2H), δ 7.81 (d, J =8.2 Hz, 2H), 7.60-7.54 (m, 1H), 7.48 (d, J =8.2 Hz, 2H), 4.11 (s, 3H), 4.10 (s , 2H), 3.70-3.37 (m, 8H). 482.22 (M+H) ⁺ 22 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.10 (s, 1H), 8.08 (s, 1H), 7.99 (s, 1H), 7.87 (d, J =8.4 Hz, 1H), 7.69 (s, 1H), 7.63 (d, J =7.8 Hz, 2H), 7.55 (d, J =8.4 Hz, 1H), 7.29 (d, J =7.8 Hz, 2H), 4.10 (s, 3H), 4.06 (s, 2H), 3.57 (s br, 4H), 2.77-2.74 (m, 2H), 2.53-2.52 (m, 2H), 2.43 (s br, 4H). 482.26 (M+H) ⁺ 23 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.17 (s, 1H), 9.00 (d, J =1.8 Hz, 1H), 8.24 (dd, J =2 ,4, 7.8 Hz, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.99 (s, 1H), 7.88 (d, J =8.4 Hz, 1H), 7.68 (d, J =8.4 Hz, 1H), 7.55 (d, J =8.4 Hz , 1H), 4.12 (s, 2H), 4.10 (s, 3H), 3.67-3.66 (m, 4H), 3.58-3.3.57 (m, 2H), 3.55-3.54 (m, 2H). 483.21 (M+H) ⁺ 24 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.13 (s, 1H), 8.09 (s, 1H), 8.00 (s, 1H), 7.88 (d, J =8.3 Hz, 1H), 7.82 (s, 1H), 7.56 (d, J =7.8 Hz, 1H), 7.38-7.31 (m, 2H), 7.25 (d, J =7.8, 1.5 Hz, 1H), δ 4.11 (s, 3H), 4.08 ( s, 2H), 3.89 (s, 3H), 3.59 (t, J =4.4, 4.4 Hz, 4H), 3.47 (s, 2H), 2.43-2.34 (m, 4H). 498.38 (M+H) ⁺ 25 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.14 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.91-7.81 (m, 2H), 7.62-7.51 (m, 3H), 7.45 (t, J =9.5, 9.5 Hz, 1H), 4.11 (s, 3H), 4.09 (s, 2H), 3.61-3.55 (m, 4H), 3.53 (s, 2H), 2, 44-2.37 (m, 4H); ¹⁹ F NMR (471 MHz, DMSO-d6, ppm) δ -118.1. 486.21 (M+H) ⁺ 26 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.88 (d, J =8.3 Hz, 1H), 7.70 (s, 1H), 7.58-7.52 (m, 2H), 7.52-7.47 (m, 1H), 7.28 (d, J =7.9 Hz, 1H), 4.11 (s, 3H), 4.07 (s, 2H), 3.59 -3.54 (m, 4H), 3.44 (s, 2H), 2.40-2.35 (m, 7H). 482.19 (M+H) ⁺ 27 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.15 (s, 1H), 8.08 (s, 1H), 8.03-7.99 (m, 2H), 7.97 (s, 1H), 7.94-7.86 (m, 2H), 7.84 (d, J =9.8 Hz, 1H), 7.57 (d, J =10.1 Hz, 1H), 4.11 (s, 3H), 4.10 (s, 2H), 3.65-3.59 (m, 6H ), 2.45-2.39 (m, 4H); ¹⁹ F NMR (471 MHz, DMSO-d6, ppm) δ -58.8 535.97 (M) ⁺ 28 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.87 (d, J =8.4 Hz, 1H), 7.73 (s, 1H), 7.68 (d, J =7.8 Hz, 2H), 7.55 (d, J =8.4 Hz, 1H), 7.28 (d, J =8.4 Hz, 2H), 4.10 (s, 3H), 4.07 (s, 2H), 3.55-3.53 (m, 4H), 3.21 (dd, J =5.4, 8.4 Hz, 1H), 2.35 (s br, 2H), 2.30-2.29 (m, 2H), 1.92-1.86 (m, 1H), 1.70-1.62 (m, 1H), 0.71 (t, J =7.2 Hz, 3H). 409.51 (M-(morpholinyl)+H) ⁺ 29 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.08 (s, 1H), 8.09 (s, 1H), 8.02 (s, 1H), 7.89 (d, J =8.3 Hz, 1H), 7.75 (s, 2H), 7.57 (d, J =8.3 Hz, 1H), 7.10 (s, 1H), 7.00 (d, J =7.9 Hz, 1H), 4.12 (s, 3H), 4.06 (s, 2H), 3.95 (s, 3H), 3.61 (t, J =4.5 Hz, 4H), 3.51 (s, 2H), 2.40 (s, 4H). 498.20 (M+H) ⁺ 30 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.29 (s, 1H), 13.18 (s, 1H), 8.84 (d, J =2.2 Hz, 1H), 8.12 (s, 1H), 8.02 (dd, J =8.3, 2.2 Hz, 1H), 7.94 (s, 1H), 7.90 (d, J =8.3 Hz, 1H), 7.84 (s, 1H), 7.55 (d, J =8.1 Hz, 1H), 7.34 (d, J =8.1 Hz, 1H), 4.05 (s, 2H). 370.15 (M+H) ⁺ 31 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.18 (s, 1H), 9.06 (d, J =2.3 Hz, 1H), 8.55 (d, J =1.8 Hz, 1H), 8.16 (t, J =2.1 Hz, 1H), 8.10 (s, 1H), 8.01 (s, 2H), 7.89 (d, J =8.4 Hz, 1H), 7.57 (d, J =8.4 Hz, 1H), 4.13 (s, 2H), 4.12 (s, 3H), 3.69 (s, 4H), 3.61 (s, 2H), 3.42 (s, 2H). 483.23 (M+H) ⁺ 32 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.15 (s, 1H), 8.10 (s, 1H), 8.02 (s, 1H), 7.89 (d, J =8.4 Hz, 1H), 7.84 (dd, J =8.0, 8.0 Hz, 1H), 7.77 (s, 1H), 7.57 (d, J =8.4 Hz, 1H), 7.29 (d, J =12.4 Hz, 1H), 7.25 (d, J =8.0 Hz , 1H), 4.12 (s, 3H), 4.11 (s, 2H), 3.60 (t, J =4.4 Hz, 4H), 3.52 (s, 2H), 2.39 (s, 4H). 486.24 (M+H) ⁺ 33 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.13 (s, 1H), 8.82 (d, J =2.4 Hz, 1H), 8.08 (s, 1H), 8.00-7.99 (m, 2H), 7.87 (d, J =8.4 Hz, 1H), 7.82 (s, 1H), 7.55 (d, J =8.4 Hz, 1H), 7.32 (d, J =7.8 Hz, 1H), 4.10 (s, 3H), 4.08 (s, 2H). 384.59 (M+H) ⁺ 34 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.14 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.87 (d, J =8.4 Hz, 1H), 7.83 (s, 1H), 7.79 (d, J =8.4 Hz, 2H), 7.55 (d, J =8.4 Hz, 1H), 7.42 (d, J =7.8 Hz, 2H), 4.10 (s, 3H), 4.09 (s, 2H), 3.58 (br, 2H), 1.62-1.60 (m, 3H), 1.55-1.48 (m, 8H). 480.60 (M+H) ⁺ 35 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.10 (s, 1H), 8.03 (s, 1H), 7.88 (d, J =8.3 Hz, 1H), 7.75-7.65 (m, 3H), 7.53 (d, J =7.9 Hz, 1H), 7.37 (d, J =8.2 Hz, 2H), 4.27 (d, J =7.2 Hz, 2H), 4.06 (s, 2H), 3.58 (t, J =4.3, 4.3 Hz, 4H), 3.48 (s , 2H), 2.40-2.34 (m, 4H), 2.33-2.25 (m, 1H), 0.90 (d, J = 6.7 Hz, 6H). 510.68 (M+H) ⁺ 36 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 7.94-7.92 (m, 2H), 7.91 (d, J =9.0 Hz, 1H), 7.69 (dd, J =0.6, 9.0 Hz, 1H), 7.67 (dd, J =0.6, 5.4 Hz, 1H), 7.59 (d, J =5.4 Hz, 1H), 7.07 (d, J =9.0 Hz, 2H), 3.90 (s, 3H). 370.14 (M+H) ⁺ 37 ^1H NMR (500 MHz, DMSO-d6, ppm) δ 13.14 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.89 (d, J =8.4 Hz, 1H), 7.86 (s, 1H), 7.83 (d, J =7.8 Hz, 1H), 7.75 (s, 1H), 7.57 (d, J =8.3 Hz, 1H), 7.52 (t, J =7.7 Hz, 1H), 7.35 (d, J =7.4 Hz, 1H), 4.12 (s, 3H), 4.10 (s, 2H), 3.67 (s, 6H), 3.41 (s, 2H). 482.02 (M+H) ⁺ 38 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.15 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.88 (d, J =8.4 Hz, 1H), 7.85 (s, 1H), 7.79 (d, J =8.4 Hz, 2H), 7.58 (d, J =8.4 Hz, 2H), 7.56 (d, J =9.0 Hz, 1H), 4.11 (s, 3H), 4.10 (s, 2H), 3.46 (dt, J =18, 6.6 Hz, 4H), 1.90-1.80 (m, 4H). 466.53 (M+H) ⁺ 39 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.12 (s, 1H), 8.09 (s, 1H), 8.00 (s, 1H), 7.88 (d, J =8.4 Hz, 1H), 7.73 (s, 1H), 7.69 (d, J =8.4 Hz, 2H), 7.56 (d, J =8.4 Hz, 1H), 7.35 (d, J =7.8 Hz, 2H), 4.11 (s, 3H), 4.08 (s, 2H), 2.65-2.61 (m, 8H). 484.40 (M+H) ⁺ 40 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.23 (s, 1H), δ 12.96 (s, 1H), 8.28 (d, J =2.6 Hz, 1H), 8.16 (d, J =7.3 Hz, 2H), 7.78 (d, J =7.4 Hz, 1H), 7.72 (dd, J =8.6, 2.5 Hz, 1H), 7.66 (d, J =8.7 Hz, 1H), 7.48 (s, 1H), 6.52-6.50 (m , 1H), 6.18 (s, 2H), 3.96 (s , 2H). 371.38 (M+H) ⁺ 41 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.22 (s, 1H), 9.28 (s, 2H), 8.11 (s, 1H), 8.10 (d, J =0.8 Hz, 1H), 8.02 (s, 1H), 7.90 (d, J =8.3 Hz, 1H), 7.57 (dd, J =8.4, 1.3 Hz, 1H), 4.16 (s, 2H), 4.12 (s, 3H), 3.70 (d, J =1.8 Hz, 4H), 3.57-3.55 (m, 2H), 3.31-3.29 (m, 2H). 484.30 (M+H) ⁺ 42 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.22-13.10 (m, 1H), 8.10 (s, 1H), 8.06 (s, 1H), 7.87 (d, J =8.2 Hz, 1H), 7.75-7.66 (m, 3H), 7.57-7.51 (m, 1H), 7.37 (d, J =8.2 Hz, 2H), 5.07-5.00 (m, 1H), 4.07 (s, 2H), 3.58 (t, J =4.4, 4.4 Hz, 4H), 3.48 (s, 2H), 2.41-2.32 (m, 4H), 1.54 (d, J =6.6 Hz, 6H). 496.55 (M+H) ⁺ 43 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.15 (s, 1H), 8.09 (s, 1H), 8.00 (s, 1H), 7.94 (s, 1H ), 7.88 (d, J =11.9 Hz, 1H), 7.57 (d, J =8.3 Hz, 1H), 7.44 (s, 1H), δ 7.32 (dd, J =7.8, 1.4 Hz, 1H), 7.25 (d, J =7.7 Hz, 1H), 4 ,12-4.09 (m, 5H), 3.92 (s, 3H), 3.56-3.49 (m, 4H), 3.22-3.12 (m, 4H). 512.38 (M+H) ⁺ 44 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.17 (s, 1H), δ8.09 (s, 1H), 8.01 (s, 1H), 7.94 (s, 1H), 7.89 (d, J =8.3 Hz, 1H), 7.75 (dd, J =10.9, 1.4 Hz, 1H), 7.63 (dd, J =7.9, 1.7 Hz, 1H), 7.56 (d, J =8.4 Hz, 1H), 7.47 (t, J =7.6, 7.6 Hz, 1H), 4.12-4.10 (m, 5H), 3.66 (s, 4H), 3.57-3.54 (m, 2H), 3.31-3.29 (m, 2H); ¹⁹ F NMR (471 MHz, DMSO-d6, ppm) δ -116.15. 500.52 (M+H) ⁺ 45 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.61 (s, 2H), 8.09 (s, 1H), 8.00 (s, 1H), 7.88 (d, J = 8.3 Hz, 1H), 7.62 (s, 1H), 7.56 (d, J = 8.2 Hz, 1H), 6.92 (s, 2H), 4.11 (s, 3H), 4.06 (s, 2H). 386.31 (M+H) ⁺ 46 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.05 (s, 1H), 8.09 (s, 1H), 7.99 (s, 1H), 7.88 (d, J =8.3 Hz, 1H), 7.56 (d, J =8.4 Hz, 1H), 7.35 (d, J =13.6 Hz, 2H), 6.69 (d, J =1.5 Hz, 1H), 4.11 (s, 3H), 4.01 (s, 2H), 3.71 (s, 3H ), 3.67-3.62 (m, 8H). 485.03 (M+H) ⁺ 47 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 8.08 (d, J = 0.6 Hz, 1H), 8.03-8.01 (m, 3H), 7.99 (s, 1H), 7.88 (d, J = 8.4 Hz, 1H), 7.77 (d, J = 9.0 Hz, 2H), 7.57 (dd, J = 0.6, 7.8 Hz, 1H), 4.14 (s, 2H), 4.11 (s, 3H), 3.66-3.64 (m, 4H), 2.92-2.90 (m, 4H). 518.49 (M+H) ⁺ 48 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.13 (s, 1H), 8.09 (s, 1H), 7.99 (s, 1H), 7.96 (s, 1H ), 7.88 (d, J =8.4 Hz, 1H), 7.81 (d, J =0.6 Hz, 1H), 7.75 (s, 1H), 7.55 (d, J =8.4 Hz, 1H), 4.11 (s, 3H), 4.07 (s, 2H), 3.70 ( sbr, 3H), 3.67-3.66 (m, 5H). 488.25 (M+H) ⁺ 49 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.14 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.88 (d, J =8.4 Hz, 1H), 7.84 (s, 1H), 7.79 (d, J =8.4 Hz, 2H), 7.56 (d, J =8.4 Hz, 1H), 7.47 (d, J =8.4 Hz, 2H), 4.11 (s, 3H), 4.10 (s, 2H), 3.00 (s, 3H), 2.97 (s, 3H). 440.37 (M+H) ⁺ 50 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.18 (s, 1H), 8.09 (s, 1H), 8.01-7.98 (m, 4H), 7.88 (d, J =8.4 Hz, 1H), 7.78 (d, J =8.4 Hz, 2H), 7.56 (d, J =8.4 Hz, 1H), 4.13 (s, 2H), 4.11 (s, 3H), 2.65 (s, 6H). 476.53 (M+H) ⁺ 51 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.12 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.87 (d, J =8.4 Hz, 1H), 7.56 (d, J =8.2 Hz, 1H), 7.46 (s, 1H), 7.22 (d, J =3.5 Hz, 1H), 6.95 (d, J =3.5 Hz, 1H), δ 4.10 (s, 3H), 4.06 (s , 2H), 3.67 (s, 2H), 3.60 (t, J =4.4 Hz, 4H), δ 2.43 (s, 4H). 474.10 (M+H) ⁺ 52 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.09 (s, 1H), δ 8.08 (d, J =0.5 Hz, 1H), 7.99 (s, 1H) , 7.87 (d, J =8.4 Hz, 1H), 7.68 (d, J =1.5 Hz, 1H), 7.58-7.54 (m, 2H), δ 7.37-7.35 (m, 1H), δ 4.10 (s, 3H), 4.04 (s, 2H), δ 3.70 (s, 2H), 3.59 (t, J =4.4 Hz, 4H), 2.44 (s, 4H). 474.19 (M+H) ⁺ 53 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.17 (s, 1H), 8.10 (s, 1H), 8.01 (s, 1H), 7.95 (s, 1H), 7.89 (d, J =8.4 Hz, 1H), 7.56 (d, J =7.7 Hz, 1H), 7.51 (d, J =8.4 Hz, 2H), 4.13-4.09 (m, 5H), 3.58-3.54 (m, 6H), 2.41 (s, 4H). 504.32 (M+H) ⁺ 54 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.13 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.89 (d, J =8.4 Hz, 1H), 7.73 (s, 1H), 7.69 (d, J =8.1 Hz, 2H), 7.56 (d, J =8.4 Hz, 1H), 7.36 (d, J =8.1 Hz, 2H), 4.11 (s, 3H), 4.07 (s, 2H), 3.58 (ddd, J =9.9, 6.2, 2.0 Hz, 2H), 3.46 (s, 2H), 2.69 (d, J =10.7 Hz, 2H), 1.66 (t, J =10.7 Hz, 2H), 1.04 (s, 3H), 1.03 (s, 3H). 496.31 (M+H) ⁺ 55 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.14 (s, 1H), 8.45 (d, J =1.5 Hz, 1H), 8.09 (s, 1H), 8.05-8.00 (m, 2H), 8.00 (s, 1H), 7.89 (d, J =8.2 Hz, 1H), 7.78-7.75 (m, 1H), 7.57 (d, J =8.2 Hz, 1H), 4.12 (s, 3H), 4.09 (s, 2H ), 3.62-3.57 (m, 4H), 3.52 (s, 2H), 2.39 (s, 4H). 469.24 (M+H) ⁺ 56 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.15 (s, 1H), 8.62 (d, J =4.9 Hz, 1H), 8.17 (s, 1H), 8.10 (s, 2H), 8.07 (s, 1H), 8.01 (s, 1H), 7.89 (d, J =8.3 Hz, 1H), 7.57 (d, J =9.0 Hz, 1H), 7.29 (dd, J =4.9, 1.2 Hz, 1H), 4 .12 (s, 5H), 3.75-3.65 (m, 5H), 3.61 (s, 3H). 483.78 (M+H) ⁺ 57 ^1H NMR (500 MHz, DMSO-d6, ppm) δ 13.06 (s, 1H), 8.07 (s, 1H), 7.98 (s, 1H), 7.86 (d, J =6.9 Hz, 1H), 7.54 (d, J =6.2 Hz, 1H), 7.30 (m, 1H), 6.18-6.13 (m, 1H), 4.10 (s, 3H), 4.01 (s, 2H), 3.93-3.88 (m, 2H), 3.62 -3.57 (m, 4H), 3.20-3.13 (m, 4H), 2.62-2.56 (m, 2H). 487.12 (M+H) ⁺ 58 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 12.99 (s, 1H), 8.07 (s, 1H), 7.97 (s, 1H), 7.86 (d, J =8.0 Hz, 1H), 7.53 (d, J =8.5 Hz, 1H), 7.11 (s, 1H), 4.10 (s, 3H), 4.05-4.03 (m, 2H), 3.96 (s, 2H), 3.05 (t, J =11.5 Hz, 1H), 2.86 (s br, 2H), 1.99 (d, J =10.5 Hz, 2H), 1.52 (qd, J =4.0, 12.5 Hz, 2H), 1.42 (s, 9H). 476.52 (M+H) ⁺ 59 ¹ H NMR (500 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.09 (s , 1H), 8.01 (s, 1H), 7.88 (d, J =8.3 Hz, 1H), 7.77-7.70 (m, 3H), 7.56 (d, J =8.1 Hz, 1H), 7.33 (d, J =8.4 Hz, 2H), 4.57 (s, 2H), 4.14 (s, 2H), 4.11 (s, 3H), 4.08 (s , 2H), 3.83 (t, J =5.2, 5.2 Hz, 2H), 3.31-3.28 (m, 2H). 482.22 (M+H) ⁺ 60 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.13 (s, 1H), 8.34 (s, 1H), 8.09 (s, 1H), 8.00 (s, 1H ), 7.88 (m, 2H), 7.55 (d, J =9.0 Hz, 1H), 4.40 (sbr, 2H), 4.11 (s, 3H), 7.08 (s, 2H), 3.73 (sbr, 6H). 489.17 (M+H) ⁺ 61 ¹ H NMR (500 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.88 (d, J =8.0 Hz, 1H), 7.72 (s, 1H), 7.69 (d, J =8.0 Hz, 2H), 7.56 (d, J =8.5 Hz, 1H), 7.29 (d, J =8.0 Hz, 2H), 4.52 (s, 2H), 4.11 (s, 3H), 4.07 (s, 2H), 3.21 (s br, 2H), 2.32 (s br, 2H), 1.74-1.72 (m, 4H). 480.28 (M+H) ⁺ 62 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.13 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.88 (d, J =7.8 Hz, 1H), 7.76 (s, 1H), 7.74 (d, J =7.8 Hz, 2H), 7.56 (d, J =8.4 Hz, 1H), 7.35 (d, J =7.8 Hz, 2H), 4.38 (s, 2H), 4.29 (t, J =7.8 Hz, 2H), 4.11 (s, 3H), 4.08 (s, 2H), 3.46 (t, J =7.8 Hz, 2H). 468.12 (M+H) ⁺ 63 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.88 (d, J =10.0 Hz, 1H), 7.77-7.69 (m, 4H), 7.56 (d, J =10.7 Hz, 1H), 7.38 (d, J =9.8 Hz, 2H), 4.11 (s, 3H), 4.08 (s, 2H), 3.57 (s, 2H), δ 3.19-3.13 (m, 2H), 2.93 (s, 2H), 2.59-2.54 (m, 2H). 481.29 (M+H) ⁺ 64 ^1H NMR (500 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.08 (s, 1H), 7.99 (s, 1H), 7.87 (d, J =8.5 Hz, 1H), 7.70 (d, J =8.0 Hz, 2H), 7.55 (d, J =9.5 Hz, 1H), 7.28 (d, J =8.0 Hz, 2H), 4.39 (s, 2H), 4.10 (s, 3H), 4.07 (s, 2H ), 3.26 (t, J =7.0 Hz, 2H), 2.31 (t, J =8.0 Hz, 2H), 1.97-1.90 (m, 2H). 466.20 (M+H) ⁺ 65 ^1H NMR (500 MHz, DMSO-d6, ppm) δ 13.09 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.87 (d, J =8.4 Hz, 1H), 7.68 (s, 1H), 7.62-7.53 (m, 3H), 7.24 (d, J =8.2 Hz, 2H), 4.11 (s, 3H), 4.06 (s, 2H), 3.23-3.09 (m, 3H), 3.07 -2.96 (m, 2H), 2.93-2.85 (m, 1H), 2.81 (s, 3H), 2.48-2.43 (m, 1H), 2.33 (s, 3H). 509.23 (M+H) ⁺ 66 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.12 (s, 1H), 8.09 (s, 1H), 8.00 (s, 1H), 7.88 (d, J =8.8 Hz, 1H), 7.75-7.00 (m, 1H), 7.68 (d, J =8.2 Hz, 2H), 7.60-7.53 (m, 1H), 7.31 (d, J =8.2 Hz, 2H), 4.11 (s, 3H), δ 4.08 (s, 2H), 4.01 (s, 2H), 3.81-3.76 (m, 2H), 3.56 (d, J =7.5 Hz, 2H), δ 3.33-3.29 (m, 2H), 2.87-2.81 (m, 2H). 496.28 (M+H) ⁺ 67 ^1H NMR (500 MHz, DMSO-d6, ppm) δ 13.10 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.88 (d, J =8.4 Hz, 1H), 7.73 (s, 1H), 7.70 (d, J =8.1 Hz, 2H), 7.57 (d, J =8.0 Hz, 1H), 7.37 (d, J =8.2 Hz, 2H), 4.11 (s, 3H), 4.07 (s, 2H), 3.64 (s, 2H), 2.88 (t, J =13.3, 13.3 Hz, 2H), 2.71 (t, J =7.0, 7.0 Hz, 2H), 2.31-2.21(m, 2H); ¹⁹ F NMR (471 MHz, DMSO-d6, ppm) δ -90.7. 488.24 (M+H) ⁺ 68 ^1H NMR (500 MHz, DMSO-d6, ppm) δ 13.64 (s, 1H), 13.23 (s, 1H), 7.95 (s, 1H), 7.90-7.79 (br, 3H), 7.75-7.62 (m, 2H), 7.60-7.49 (m, 2H), 4.48-4.29 (m, 2H), 4.06 (s, 2H), 4.02-3.91 (m, 2H), 3.73-3.57 (m, 2H) , 3.21-3.05 (m, 2H). 532.31 (M+H) ⁺ 69 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.08 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.88 (d, J =8.3 Hz, 1H), 7.65 (d, J =8.7 Hz, 1H), 7.61 (s, 1H), 7.56 (d, J =8.3 Hz, 1H), 7.01 (d, J =8.7 Hz, 1H), 4.14 (t, J =5.8 Hz, 2H), 4.11 (s, 3H), 4.05 (s, 2H), 3.61- 3.57 (m, 4H), 2.71 (t, J =5.8 Hz, 2H), 2.49 (s, 4H). 498.9 (M+H) ⁺ 70 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.12 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.89 (d, J =8.4 Hz, 1H), 7.82 (s, 1H), 7.57 (d, J =7.8 Hz, 1H), 7.34 (t, J =7.9 Hz, 1H), 7.32 (s, 1H), 7.28 (d, J =7.6 Hz, 1H), 6.91 (dd, J =7.8, 1.9 Hz, 1H), 4.19 (t, J =5.8 Hz, 2H), 4.12 (s, 3H), 4.08 (s, 2H), 3.62-3.59 (m, 4H), 2.73 (t, J =5.8 Hz, 2H). 498.21 (M+H) ⁺ 71 ^1H NMR (500 MHz, CDCl ₃ ) δ 8.00 (d, J =0.8 Hz, 1H), 7.75 (d, J =8.4 Hz, 1H), 7.66 (s, 1H ), 7.33 (dd, J =8.4, 1.2 Hz, 1H), 7.25 (s, 1H), 3.97 (s, 3H), 3.81 - 3.76 (m, 4H), 3.62 (s, 2H), 3.55 (s, 2H), 2.68-2.63 (m, 4H). 416.89 (M+H) ⁺ 72 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.04 (s, 1H), 8.09 (s, 1H), 8.00 (s, 1H), 7.87 (d, J =8.1 Hz, 1H), 7.57 (s, 1H), 7.44 (t, J =8.7 Hz, 2H), 7.42 (s, 1H), 6.67 (d, J =8.7 Hz, 2H), 5.82 (d, J =8.2 Hz, 1H), 4.11 (s, 3H), 4.02 (s, 2H), 3.91-3.86 (m, 2H), 3.52-3.47 (m, 1H), 3.44 (td, J =11.5, 2.1 Hz, 3H), 1.90 (d, J =13.3 Hz, 2H), 1.43-1.35 (m, 2H). 468.76 (M+H) ⁺ 73 ¹ H NMR (500 MHz, DMSO-d6, ppm) δ 13.06 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.88 (d, J =8.6 Hz, 1H), 7.58 - 7.51 (m, 3H), 7.49 (s, 1H), 6.89 (d, J =8.8 Hz, 2H), 4.12 (s, 3H), 4.03 (s, 2H), 3.94 (d, J =10.1 Hz, 3H), 3.48 (t, J =11.0 Hz, 2H), 2.78 (s, 3H), 1.77 (ddd, J =16.1, 12.3, 4.5 Hz, 2H), 1.59 (d, J = 10.1 Hz, 2H). 482.39 (M+H) ⁺ 74 ¹ H NMR (500 MHz, DMSO-d6, ppm): d 12.65 (s, 1H), 9.31 (s, 1H), 7.68-7.66 (m, 3H), 7.35 (d, J = 8.5 Hz, 2H), 7.00 (s, 1H), 6.86 (dd, J = 2.0, 8.0 Hz, 1H), 6.72 (d, J = 8.0 Hz, 1H), 3.84 (s, 2H), 3.59-3.57 (m, 4H), 3.48 (s, 2H), 2.37-2.36 (m, 4H). 482.77 (M+H) ⁺ 445.33 [M-(CH)+H ⁺ ] 75 ¹ H NMR (500 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.89 (d, J =8.3 Hz, 1H), 7.73 (s, 1H), 7.69 (d, J =8.2 Hz, 2H), 7.57 (d, J =7.5 Hz, 1H), 7.38 (d, J =8.2 Hz, 2H), 4.12 (s, 3H), 4.08 (s, 2H), 3.99 (d, J =13.7 Hz, 1H), 3.65 (dd, J =11.2, 3.2 Hz, 2H), 3.49-3.42 (m, 1H), 3.20-3.14 (m, 2H), 2.16-2.09 (m, 1H), 1.04 (d, J =6.2 Hz, 3H). 482.93 (M+H) ⁺ 76 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.89 (d, J =8.3 Hz, 1H), 7.74 (s, 1H), 7.70 (d, J =8.2 Hz, 2H), 7.57 (d, J =8.6 Hz, 1H), 7.30 (d, J =8.3 Hz, 2H), 6.36 (s, 1H), 4.12 (s, 3H), 4.08 (s, 2H), 3.70 (t, J =5.5 Hz, 2H), 3.61 (t, J =5.6 Hz, 2H), 2.37 (t, J =5.0 Hz, 2H). 465.17 (M+H) ⁺ 77 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.10 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.89 (d, J =8.4 Hz, 1H), 7.70 (s, 1H), 7.65 (d, J =8.1 Hz, 2H), 7.57 (d, J =8.1 Hz, 1H), 4.11 (s, 3H), 4.07 (s, 2H), 3.83 (dd, J =11.4, 2.6 Hz, 2H), 3.25 (td, J =11.7, 1.9 Hz, 2H), 2.55 (d, J =7.1 Hz, 2H), 1.78-1.74 (m, 1H), 1.51 (d, J =13.0 Hz, 2H), 1.26-1.19 (m, 2H). 467.19 (M+H) ⁺ 78 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 12.9 (s, 1H), 8.07 (s, 1H), 7.97 (s, 1H), 7.86 (d, J = 8.4 Hz, 1H), 7.53 (d, J = 8.3 Hz, 1H), 6.45 (s, 1H), 4.09 (s, 3H), 3.89 (s, 2H), 3.78-3.73 (m, 4H), 3.13-3.09 (m, 4H). 377.70 (M) ⁺ 79 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.0 (s, 1H), 8.08 (s, 1H), 7.98 (s, 1H), 7.86 (d, J = 8.2 Hz, 1H), 7.55 (d, J = 7.7 Hz, 1H), 7.05 (s, 1H), 4.34 (s, 2H), 4.10 (s, 3H), 4.09-4.06 (m, 2H), 3.99-3.95 (m, 4H). 392.00 (M+H) ⁺ 80 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.88 (d, J =8.4 Hz, 1H), 7.72 (s, 1H), 7.68 (d, J =8.4 Hz, 2H), 7.56 (d, J =8.4 Hz, 1H), 7.34 (d, J =7.8 Hz, 2H), 4.11 (s, 3H), 4.07 (s, 2H), 3.56-3.54 (m, 4H), 3.46 (s, 2H), 2.86-2.84 (m, 2H), 2.44 (s br, 4H), 2.10-2.08 (m, 1H), 1.9-1.91 (m, 2H), 1.74-1.72 (m, 2H), 1.39 (dq, J = 3.0, 12 Hz, 2H). 550.89 (M) ⁺ 81 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.12 (s, 1H), 12.85 (s, 1H), 7.87-7.79 (m, 2H), 7.76-7.66 (m, 3H), 7.51 (d, J =8.3 Hz, 1H), 7.38 (d, J =8.0 Hz, 2H), 4.05 (d, J =27.7 Hz, 2H), 3.60 (d, J =4.1 Hz, 4H), 3.49 (s, 2H), 2.39 (dt, J =3.7, 1.8 Hz, 4H). 468.19 (M+H) ⁺ 82 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), δ 8.52 (d, J =2.2 Hz, 1H), 8.09 (s, 1H) , δ 8.00 (s, 1H), 7.92-7.85 (m, 2H), 7.63 (s, 1H), δ 7.60 (d, J =8.2 Hz, 1H), 6.93 (d, J =8.8 Hz, 1H), 4.10 (s, 3H), 4, 06 (s, 2H), 3.74-3.70 (m, 4H), 3.52-3.48 (m, 4H). 454.92 (M) ⁺ 83 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.28 (s, 1H), 13.14 (s, 1H), 8.11 (s, 1H), 7.92 (s, 1H ), 7.88 (d, J =8.4 Hz, 1H), 7.71 (s, 1H), 7.67 (d, J =8.4 Hz, 2H), 7.53 (d, J =8.4 Hz, 1H), 7.33 (d, J =8.4 Hz, 2H), 4.02 (s , 2H), 3.46 (s, 2H), 2.39-2.25 (s br, 8H), 2.14 (s, 3H). 467.04 (M+H) ⁺ 84 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.12 (s, 1H), 8.08 (s, 1H), 7.99 (s, 1H), 7.87 (d, J =8.4 Hz, 1H), 7.72 (s, 1H), 7.67 (d, J =7.8 Hz, 2H), 7.55 (d, J =7.2 Hz, 1H), 7.34 (d, J =8.4 Hz, 2H), 4.10 (s, 3H), 4.07 (s, 2H), 3.46 (s, 2H), 2.39-2.39 (br, 8H), 2.17 (s, 3H). 481.01 (M) ⁺ 85 ¹ H NMR (600 MHz, DMSO-d ⁶ , ppm) δ 13.11 (s, 1H), δ 8.49 (d, J =2.4 Hz, 1H), δ 8.09 (s, 1H), δ 8.00 (s, 1H), δ 7.89-7.85 (m, 2H), δ 7.61 (s, 1H), δ 7.55 (d, J =8.4 Hz, 1H), δ 6.91 (d, J =8.8 Hz, 1H), δ 4, 11 (s, 3H), δ 4.05 (s, 2H), δ 3.56-3.52 (m, 4H), δ 2.42-2.39 (m, 4H), δ 2.22 (s, 3H). 468.42 (M+H) ⁺ 86 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.22 (s, 1H), 8.41 (d, J =5.3 Hz, 1H), 8.20 (s, 1H), 8.09 (s , 1H), 8.01 (s , 1H), 7.90-7.87 (m, 2H), 7.74 (dd, J =5.3, 1.6 Hz, 1H), 7.56 (d, J =8.9 Hz, 1H), 4.16 (s, 2H), 4.11 (s, 3H). 404.19 (M+H) ⁺ 87 ^1H NMR (600 MHz, MeOH-d4, ppm) δ 8.16 (d, J =5.3 Hz, 1H), 8.02 (s, 1H), 7.86 (s, 1H), 7.85 (s, 1H), 7.78 (s, 1H), 7.54 (d, J = 8.6 Hz, 1H), δ 7.14 (s, 1H), 7.07-7.05 (m, 1H), 4.74-4.49 (m, 4H), 4.12 (s, 3H), 4, 00 (s, 2H), 3.41-3.32 (m, 4H), 2.94 (s, 3H). 468.26 (M+H) ⁺ 88 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.14 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.87 (d, J =8.5 Hz, 1H), 7.81 (s, 1H), 7.67-7.64 (m, 1H), 7.55 (d, J =8.0 Hz, 1H), 7.30-7.27 (m, 1H), 4.11 (s, 2H), 4.10 (s, 3H), 3.58 -3.57 (m, 6H), 2.41 (s br, 4H); ¹⁹ F NMR (471 MHz, DMSO-d6, ppm): d -141.0 (d), -142.5 (d). 504.54 (M+H) ⁺ 89 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.23 (s, 1H), 8.19-8.12 (m, 2H), 7.83 (s, 1H), 7.74-7.67 (m, 3H), 7.37 (d, J = 8.1 Hz, 2H), 4.36 (s, 3H), 4.09 (s, 2H), 3.59-3.57 (m, 4H), 3.48 (s, 2H), 2.37 (s, 4H). 469.16 (M+H) ⁺ 90 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.17 (s, 1H), 8.09 (s, 1H), 8.00 (s, 1H), 7.90-7.84 (m, 2H), 7.75 (dd, J =8.1, 1.7 Hz, 1H), 7.66 (s, 1H), 7.63 (d, J =8.1 Hz, 1H), 7.56 (d, J =7.6 Hz, 1H), 4.11 (s, 3H), 4.09 (s, 2H), 3.61-3.57 (m, 4H), 3.55 (s, 2H), 2.40 (s, 4H). 552.36 (M+H) ⁺ 91 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.11 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.88 (d, J =8.3 Hz, 1H), 7.56 (d, J =8.3 Hz, 1H), 7.41 (d, J =7.8 Hz, 1H), 7.37 (s, 1H), 7.27 (s, 1H), 7.21 (d, J =8.5 Hz, 1H), 4.10 (s, 3H ), 4.07 (s, 2H), 3.58 (t, J =4.5 Hz, 4H), 3.45 (s, 2H), 2.46 (s, 3H), 2.37 (s, 4H). 482.24 (M+H) ⁺ 92 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.07 (s, 1H), 8.28 (d, J =2.4 Hz, 1H), 8.08 (s, 1H), 7.99 (s, 1H), 7.87 (d, J =8.4 Hz, 1H), 7.72 (dd, J =2.4, 8.4 Hz, 1H), 7.56 (d, J =7.2 Hz, 1H), 7.49 (s, 1H), 6.51 (d, J =8.4 Hz , 1H), 6.19 (s, 2H), 4.10 (s, 3H), 4.03 (s, 2H). 385.20 (M+H) ⁺ 93 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.06 (s, 1H), 8.09 (s, 1H), 8.00 (s, 1H), 7.88 (d, J =8.4 Hz, 1H), 7.62 (d, J =5.0 Hz, 1H), 7.56 (dd, J =8.4, 1.0 Hz, 1H), 7.32 (d, J =5.0 Hz, 1H), 4.11 (s, 3H), 4.03 (s, 2H). 427.25 (M+H) ⁺ 94 1H NMR (600 MHz, DMSO-d6, ppm) δ 13.12 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.88 (d, J =8.4 Hz, 1H), 7.72 (s, 1H), 7.68 (d, J =8.4 Hz, 2H), 7.56 (d, J =7.8 Hz, 1H), 7.35 (d, J =7.8 Hz, 2H), 4.11 (s, 3H), 4.07 (s, 2H), 3.41 (s, 2H), 2.16 (s, 6H). 426.29 (M+H) ⁺ 95 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.14 (s, 1H), 8.45 (s, 1H), 8.10 (s, 1H), 8.03 (s, 1H), 8.01 (d, J =8.1 Hz, 2H), 7.89 (d, J =8.1 Hz, 1H), 7.77 (d, J =8.2 Hz, 1H), 7.57 (d, J =8.2 Hz, 1H), 4.12 (d, J =3.1 Hz, 3H), 4.10 (s, 2H), 3.61-3.58 (m, 4H), 3.52 (s, 2H), 2.40 - 2.38 (m, 4H). 469.65 (M+H) ⁺ 96 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.03 (s, 1H), 8.08 (s, 1H), 7.99 (s, 1H), 7.86 (d, J =8.5 Hz, 1H), 7.54 (d, J =8.0 Hz, 1H), 7.13 (s, 1H), 4.10 (s, 3H), 3.98 (s, 2H), 3.21 (tt, J =3.5, 12 Hz, 2H), 3.01 (td, J =2.5, 12.5 Hz, 2H), 2.16 (d, J =13 Hz, 2H), 1.84 (qd, J =3.5, 12.5 Hz, 2H). 376.06 (M+H) ⁺ 97 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.10 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.88 (d, J =8.5 Hz, 1H), 7.71 (s, 1H), 7.68 (d, J =8.0 Hz, 2H), 7.56 (d, J =8.5 Hz, 1H), 7.38 (d, J =8.0 Hz, 2H), 4.11 (s, 3H), 4.07 (s, 2H), 3.71 (t, J =6.0 Hz, 2H), 3.64 (s, 2H), 3.63-3.61 (m, 2H), 2.65-2.61 (m, 4H), 1.82 (q, J = 6.0 Hz, 2H). 482.19 (M+H) ⁺ 98 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.01 (s, 1H), 8.08 (s, 1H), 7.98 (s, 1H), 7.86 (d, J =8.4 Hz, 1H), 7.53 (d, J =8.4 Hz, 1H), 7.11 (s, 1H), 4.10 (s, 3H), 3.96 (s, 2H), 3.59 (sbr, 4H), 3.18-3.11 (m, 4H), 3.05 (d, J =13.2 Hz, 2H), 2.71-2.67 (m, 2H), 2.41-2.40 (m, 4H), 2.07-2.02 (m, 2H), 1.68-1.65 (m, 1H), 1.47-1.44 (m, 1H). 503.42 (M+H) ⁺ 99 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.13 (s, 1H), 8.09 (s, 2H), 8.01 (s, 1H), 7.88 (d, J =8.2 Hz, 1H), 7.70 (d, J =3.0 Hz, 1H), 7.68 (d, J =8.2 Hz, 2H), 7.58 (d, J =8.3 Hz, 1H), 7.37 (d, J =8.3 Hz, 2H), 4.11 (s, 3H), 4.08 (s, 2H), 3.60 (s, 2H), 2.46 (s, 4H), 1.72 (dd, J =6.7, 3.3 Hz, 4H). 452.19 (M+H) ⁺ 100 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.10 (sbr, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.87 (d, J =8.0 Hz, 1H), 7.69 (s, 1H), 7.67 (d, J =8.0 Hz, 2H), 7.57 (d, J =8.5 Hz, 1H), 7.34 (d, J =8.0 Hz, 2H), 4.11 (s, 3H), 4.07 (s, 2H), 2.34 (sbr, 4H), 1.53-1.48 (m, 4H), 1.39 (sbr, 2H). 466.09 (M+H) ⁺ 101 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.36-12.95 (br, 1H), δ 8.08 (s, 1H), δ 8.01 (s, 1H), δ 7.87 (d, J =8.3 Hz, 1H), δ 7.75-7.69 (m , 3H), δ 7.60-7.54 (m, 1H), δ 7.32 (d, J =8.0 Hz, 2H), δ 4.53 (s, 2H), δ 4.11 ( s, 3H), δ 4.08 (s, 2H), δ 3.31 (s, 3H), δ 3.19 (t, J =5.2 Hz, 2H), δ 2.87 (t, J =5.2 Hz, 2H). 481.20 (M+H) ⁺ 102 ¹ H NMR (600 MHz, DMSO-d6, ppm) δ 13.12 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.88 (d, J =8.3 Hz, 1H), 7.71 (s, 1H), 7.67 (d, J =8.2 Hz, 2H), 7.58 (s, 1H), 7.30 (d, J =8.2 Hz, 2H), 4.62 (s, 4H), 4.12 (s, 3H), 4.08 (s, 2H), 3.51 (s, 2H), 3.30 (s, 4H). 480.92 (M+H) ⁺ 103 ^1H NMR (600 MHz, DMSO-d6, ppm) δ 13.12 (s, 1H), 8.09 (s, 1H), 8.01 (s, 1H), 7.88 (d, J =8.4 Hz, 1H), 7.74-7.66 (m, 4H), 7.56 (d, J =7.8 Hz, 1H), 7.33 (d, J =7.8 Hz, 1H), 4.11 (s, 3H), 4.07 (s, 2H), 3.94-3.91 (m, 1H ), 3.58 (sbr, 1H), 3.19 (sbr, 2H), 2.99-2.95 (m, 1H), 2.04-1.99 (m, 1H). 438.3 (M+H) ⁺ 104 ¹ H NMR (500 MHz, DMSO-d6, ppm) δ 13.10 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.88 (d, J =8.0 Hz, 1H), 7.73 (s, 1H), 7.69 (d, J =8.5 Hz, 2H), 7.56 (d, J =8.0 Hz, 1H), 7.36 (d, J =8.0 Hz, 2H), 4.11 (s, 3H), 4.07 (s, 2H), 3.74 (s, 2H), 3.62 (t, J =12.5 Hz, 4H); ¹⁹ F NMR (471 MHz, _CDCl3 , ppm) δ -97.59. 474.29 (M+H) ⁺ 105 ^1H NMR (500 MHz, DMSO-d6, ppm) δ 13.10 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.88 (d, J =9.0 Hz, 1H), 7.71 (s, 1H), 7.67-7.64 (m, 3H), 7.56 (d, J =8.4 Hz, 1H), 7.32-7.31 (m, 2H), 4.11 (s, 3H), 4.07 (s, 2H), 3.59 (s, 2H), 3.50-3.48 (m, 2H), 3.25 (m, 2H), 2.86-2.85 (m, 2H). 468.28 (M+H) ⁺

EXAMPLE 2: Evaluation of the inhibitory activity of compounds of formula (I) in in vitro biochemical assays

Various compounds of Formula I were tested for their inhibitory activity against a variety of receptor tyrosine kinases. Brief descriptions of the various assays are provided below.

FLT3 kinase assay

Inhibition of FLT3 kinase activity by the test compound disclosed herein was assessed using the AlphaScreen assay (PerkinElmer, Akron, OH, USA). Standard assay conditions were 1.5 μg recombinant FLT3 kinase (SignalChem, Richmond, BC) with 6 μg biotin-conjugated Poly-(Glu 4: Tyr 1) (Cisbio, Bedford, MA, USA) in assay buffer (10 μM ATP, 10 mM potassium chloride, pH 7.0, 0.21 mM EDTA, 0.5% glycerol, 1 mg/mL BSA, 0.01% 2-mercaptoethanol, 0.001% Brij35, 10 mM _MgCl2 ) in a final volume of 25 μL. Reactions were incubated at 30 °C for 30 min and the incubation was stopped by adding 5 μl of 50 mM EDTA. The resulting product was analyzed using the AlphaScreen kit and counted using the Enspire Alpha (PerkinElmer, Waltham, MA, USA). Readings from the control reaction (complete reaction mixture) were designated as 0% inhibition and readings from the reaction without enzyme were designated as 100% inhibition. IC50 values of the inhibitors were determined after running assays with each compound at serially diluted concentrations in duplicate. Results were analyzed using linear regression software (GraphPad Prism 5; GraphPad Software Inc.)

FLT3-D835Y mutant kinase assay

The inhibition of FLT3-D835Y mutant kinase activity by the test compound disclosed herein was assessed using AlphaScreen (PerkinElmer). Standard assay conditions were 1 μg recombinant FLT3-D835Y mutant kinase (SignalChem) with 6 μg biotin-conjugated Poly-(Glu 4: Tyr 1) (Cisbio) in assay buffer (10 μM ATP, 10 mM potassium chloride, pH 7.0, 0.21 mM EDTA, 0.5% glycerol, 1 mg/mL BSA, 0.01% 2-mercaptoethanol, 0.001% Brij35, 10 mM MgCl2 ₎ in a final volume of 25 μL. Reactions were incubated at 30°C for 30 min and the incubation was stopped by adding 5 μl of 50 mM EDTA. The resulting product was analyzed using the AlphaScreen kit and counted using the Enspire Alpha (PerkinElmer). Readings from the control reaction (complete reaction mixture) were designated as 0% inhibition and readings from the reaction without enzyme were designated as 100% inhibition. IC ₅₀ values of the inhibitors were determined after running assays with each compound at serially diluted concentrations in duplicate. Results were analyzed using linear regression software (GraphPad Prism 5; GraphPad Software Inc.)

αPDGFR kinase assay

The inhibition of PDGFRα kinase activity by the test compound disclosed herein was assessed using AlphaScreen (PerkinElmer). The standard assay conditions were 1.5 μg recombinant PDGFRα kinase (SignalChem) with 6 μg biotin-conjugated Poly-(Glu 4:Tyr 1) (Cisbio) in assay buffer (10 μM ATP, 10 mM potassium chloride, pH 7.0, 0.21 mM EDTA, 0.5% glycerol, 1 mg/mL BSA, 0.01% 2-mercaptoethanol, 0.001% Brij35, 10 mM _MgCl2 ) in a final volume of 25 μL. Reactions were incubated at 30°C for 30 min and the incubation was stopped by adding 5 μl of 50 mM EDTA. The resulting product was analyzed using the AlphaScreen kit and counted using the Enspire Alpha (PerkinElmer). Readings from the control reaction (complete reaction mixture) were designated as 0% inhibition and readings from the reaction without enzyme were designated as 100% inhibition. IC ₅₀ values of the inhibitors were determined after running assays with each compound at serially diluted concentrations in duplicate. Results were analyzed using linear regression software (GraphPad Prism 5; GraphPad Software Inc.)

MET kinase assay

The compounds disclosed herein were tested for MET kinase activity inhibition as assessed using AlphaScreen (PerkinElmer) as a reverse screen to demonstrate their selective kinase inhibition activity against class III receptor tyrosine kinases. Since MET kinase belongs to the class VIII receptor tyrosine kinase family, it has one transmembrane α chain associated with one extracellular β chain, which differs from the class III RTK family with 5 immunoglobulin-like extracellular domains. Standard assay conditions were 1 μg recombinant MET kinase (SignalChem) with 3 μg biotin-conjugated Poly-(Glu 4: Tyr 1) (Cisbio) in assay buffer (10 μM ATP, 10 mM calcium chloride, pH 7.0, 0.21 mM EDTA, 0.5% glycerol, 1 mg/ml BSA, 0.01% 2-mercaptoethanol, 0.001% Brij35, 10 mM MgCl ₂ ) in a final volume of 25 μl. Reactions were incubated at 30°C for 45 min, and inhibition was terminated by the addition of 5 μl 50 mM EDTA. The resulting product was analyzed using the AlphaScreen kit and counted using the Enspire Alpha (PerkinElmer). Readings for the control reaction (complete reaction mixture) were designated as 0% inhibition, and readings for the reaction without enzyme were designated as 100% inhibition. IC50 values for the inhibitors were determined after running assays with each compound at serially diluted concentrations in duplicate. Results were analyzed using linear regression software (GraphPad Prism 5; GraphPad Software Inc.)

Results

In comparison with the MET kinase inhibition activity, the compounds of the invention were found to have an unexpected and selective inhibitory activity against class III RTKs. More specifically, all the compounds of the invention show IC ₅₀ values below 10 nM for FLT3 kinase as well as for the mutant FLT3-D835Y kinase, and 57 out of 105 compounds of the invention show an IC ₅₀ value below 10 nM for PDGFR kinase, which demonstrates the high specific activity of the compounds of the invention against class III RTKs. The IC ₅₀ values for the compounds of the invention against mutant FLT3 kinases, FLT3-D835Y, PDGFRα kinase and Met kinase are summarized in Table 3. In the table, the symbol "A+" means that the IC ₅₀ is less than 1 nM, "A" means that the IC ₅₀ is in the range of 1-10 nM, "B" means that the IC ₅₀ is in the range of 10-30 nM, "C" means that the IC ₅₀ is from 30 to 100 nM, "D" means that the IC ₅₀ is from 100 to 300 nM, "E" means that the IC ₅₀ is from 300 to 1000 nM and "F" means that the IC ₅₀ is greater than 1000 nM.

Table 3. Ranking of IC ₅₀ values for individual receptor tyrosine kinases

Compound
No. FLT3 FLT3-
D835Y PDGFRa MET 1 A A+ B F 2 A A+ B F 3 A+ A+ A F 4 A A+ B F 5 A A B F 6 A+ A+ A F 7 A A A F 8 A+ A A F 9 A+ A+ A F 10 A+ A+ A F 11 A A+ B F 12 A A C F 13 A A B F 14 A A B F 15 A+ A+ A F 16 A+ A+ A F 17 A A B F 18 A+ A A F 19 A A A F 20 A A A F 21 A+ A+ A F 22 A+ A B F 23 A+ A+ A F 24 A+ A+ A F 25 A A A F 26 A A B F 27 A A D F 28 A A B F 29 A+ A+ B F 30 A+ A+ A F 31 A+ A+ D F 32 A+ A+ B F 33 A+ A+ A F 34 A A C F 35 A A C F 36 A A+ A F 37 A A A F 38 A A B F 39 A A B F 40 A A+ A F 41 A+ A B F 42 A+ A B F 43 A A A F 44 A+ A+ A F 45 A+ A+ B F 46 A+ A+ A F 47 A+ A+ A F 48 A+ A+ A F 49 A+ A+ A F 50 A+ A+ B F 51 A+ A+ B F 52 A+ A+ B F 53 A+ A C F 54 A+ A+ A F 55 A+ A+ A F 56 A A A F 57 A+ A+ B F 58 A A C F 59 A+ A+ A F 60 A+ A+ A F 61 A A+ A F 62 A A+ B F 63 A A A F 64 A A B F 65 A A+ A F 66 A+ A+ F F 67 A A B F 68 A A A F 69 A+ A+ B F 70 A A+ B F 71 A A+ C F 72 A A B F 73 A A C F 74 A+ A+ A F 75 A+ A+ A F 76 A A D F 77 A A C F 78 A+ A+ A F 79 A+ A+ A F 80 A+ A+ A E 81 A A A F 82 A A D F 83 A+ A+ A E 84 A+ A+ A F 85 A+ A+ A E 86 A A B F 87 A+ A+ A E 88 A A C F 89 A A A F 90 A A D F 91 A A C F 92 A+ A+ A F 93 A A D F 94 A+ A A E 95 A+ A+ A F 96 A+ A+ A F 97 A+ A+ C F 98 A+ A+ A F 99 A+ A+ A F 100 A+ A+ A D 101 A+ A+ A F 102 A+ A+ A E 103 A+ A+ A E 104 A A C F 105 A+ A+ A E

EXAMPLE 3: Evaluation of the anti-proliferative activity of compounds of formula (I) against a panel of human cancer cell lines with different signal dependence

As noted above, the compounds of the invention can be used to treat diseases or disorders associated with protein kinase. Diseases associated with protein kinase can be cancer, an autoimmune disease or a proliferative disorder of blood vessels. Cancer can be lung cancer, colon cancer, colorectal cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer, bladder cancer, stomach cancer, kidney cancer, salivary gland cancer, ovarian cancer, endometrial cancer, cervical cancer, oral cancer, skin cancer, brain cancer, lymphoma or leukemia.

Cell growth inhibition by the compounds was measured using the CellTiterTM-96 assay. The cytotoxicity of the compounds of the invention was assessed in panels of cell lines with different signaling pathway dependencies, including the MV4-11 human acute myeloid leukemia cell with the FLT3-ITD mutation, the Molm-14 human acute myeloid leukemia cell with the FLT3-ITD mutation, the THP-1 human acute monocytic leukemia cell with the NRAS G12D mutation, the A375 melanoma cell with the BRAF V600E mutation, and the HCC827 NSCLC cell with the EGFR exon 19 deletion mutation.

MV4-11 cell culture

The compounds disclosed herein were tested in a MV4-11 cell proliferation assay to determine their specific cellular activity against FLT3 signaling pathway-dependent cell growth. MV4-11 cells are human acute myeloid leukemia (AML) cells containing a ligand-independent FLT3-ITD activating mutation that renders abnormal AML cell proliferation dependent on the FLT3 signaling pathway. Inhibition of FLT3 kinase activity will result in decreased cell growth and/or cell death. MV4-11 cells (Catalog #CRL-9591) were obtained from the American Type Culture Collection (ATCC, Manassas, VA). Briefly, cells were grown in IMDM supplemented with 10% heat-inactivated fetal bovine serum and incubated at 37°C in a 5% _CO2 atmosphere. Cells were allowed to expand until saturation was reached, after which they were subcultured or harvested for use in the assay.

Molm-14 cell culture

The compounds disclosed herein were tested in a Molm-14 cell proliferation assay to determine their specific cellular activity against FLT3 signaling pathway-dependent cell growth. Molm-14 cells are human acute myeloid leukemia (AML) cells containing a ligand-independent FLT3-ITD activating mutation that renders abnormal AML cell proliferation dependent on the FLT3 signaling pathway. Inhibition of FLT3 kinase activity will result in decreased cell growth and/or cell death. Molm-14 cells (catalog #ACC-777) were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany). Briefly, cells were grown in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum and incubated at 37°C in 5% _CO2 . Cells were allowed to expand until saturation was reached, after which they were subcultured or harvested for use in the assay.

THP-1 cell culture

The compounds disclosed herein were tested in a THP-1 cell proliferation assay as a reverse screen to demonstrate their selective anti-proliferative activity against cell growth that is entirely dependent on class III RTK signaling. THP-1 cells are human acute monocytic leukemia cells with the NRAS G12D mutation, which renders cell growth independent of class III RTK signaling. THP-1 cells (catalog #TIB-202) were obtained from ATCC. Briefly, cells were grown in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum and 0.05 mM 2-mercaptoethanol and incubated at 37°C in an atmosphere of 5% CO ₂ . Cells were allowed to expand until saturation was reached, after which they were subcultured or harvested for use in the assay.

Cell culture A375

The compounds disclosed herein were tested in an A375 cell proliferation assay as a reverse screen to demonstrate their selective anti-proliferative activity against cell growth that is entirely dependent on class III RTK signaling. A375 cells are melanoma cells encoding the BRFA V600E mutation, which renders cell growth dependent on BRAF signaling instead of class III RTK signaling. A375 cells (Catalog #CRL-1619) were obtained from ATCC. Briefly, cells were cultured in DMEM containing 10% FBS and incubated at 37°C under 5% CO ₂ . Cells were allowed to expand to saturation and were then subcultured or harvested for use in the assay.

HCC827 cell culture

The compounds disclosed herein were tested in a HCC827 cell proliferation assay as a reverse screen to demonstrate their selective anti-proliferative activity against cell growth that is entirely dependent on class III RTK signaling. HCC827 cells are non-small cell lung carcinoma (NSCLC) cells encoding an EGFR exon 19 deletion mutation that renders cell growth dependent on EGFR signaling instead of class III RTK signaling. HCC827 cells (catalog #CRL-2868) were obtained from ATCC. Briefly, cells were cultured in RPMI 1640 medium supplemented with 10% FBS and incubated at 37°C under 5% CO ₂ . Cells were allowed to expand to saturation before being subcultured or harvested for use in the assay.

MV4-11, Molm-14, THP-1, A375 and HCC827 cells were seeded in 96-well plates at 6500, 25000, 5000, 2000 and 8000 cells/well, respectively, and incubated at 37°C under 5% _CO2 overnight. The seeded cells were then treated with increasing concentrations of the test compounds and incubated for an additional 72 h. At the end of the incubation, CellTiter96® AQueous One Solution reagent (Promega, San Luis Obispo, CA, USA) was added and incubated for an additional 2-4 h. Cell viability was determined by measuring the optical density at 490 nm using an EMax® microplate reader (Molecular Devices, San Jose, CA, USA).

Results

In comparison with the anti-proliferative activity in THP-1, A375 or HCC827 cells, the compounds of the invention showed unexpected and selective cytotoxicity towards cell growth that is entirely dependent on the class III RTK signaling system. More specifically, 68 compounds of the invention show IC ₅₀ values below 30 nM for the growth of leukemia cells dependent on the FLT3 signaling system, demonstrating the high specific cellular activity of the compounds of the invention.

The 50% inhibitory concentration (IC ₅₀ ) values were calculated by plotting the percentage growth inhibition against the compound concentration using GraphPad Prism 5 curve fitting software. The anti-proliferative activity of the compounds of the invention is summarized in Table 4. In the table, the symbol "A" means that the IC ₅₀ is less than 0.03 μM, "B" means that the IC ₅₀ is from 0.03 to 0.1 μM, "C" means that the IC ₅₀ is from 0.1 to 0.3 μM, "D" means that the IC ₅₀ is from 0.3 to 1 μM, "E" means that the IC ₅₀ is from 1 to 10 μM, and "F" means that the IC ₅₀ is greater than 10 μM.

Table 4. Ranking of anti-proliferative activity against cell line panel

Compound
No. MV4-11
(FLT3-ITD) Molm-14
(FLT3-ITD) THP-1
NRAS ^N12D A375
BRAF ^V600E HCC827
EGFR ^Exon19_del 1 B B F F F 2 B B F F F 3 A A F F F 4 C C F F F 5 B C F F F 6 A A F F F 7 A A F F F 8 A B F F F 9 A A F F F 10 A A F F F 11 A B F F F 12 B C F F F 13 B B F F F 14 A B F E F 15 A A F F F 16 A A F F F 17 A B F F F 18 A A F F F 19 A A F F F 20 A B F F F 21 A A F F F 22 A B F F F 23 A B F F F 24 A B F F F 25 A B F F F 26 C C F F F 27 D E F F F 28 B B F F F 29 A B F F F 30 B B F F F 31 A A E E E 32 B C F F F 33 B B F F F 34 B B F F F 35 C C E F F 36 A B F F F 37 A B F F F 38 A B F F F 39 B B F F F 40 A A E E E 41 A A F F F 42 B C F F F 43 B B F F F 44 A A F F F 45 A A F F F 46 A A F F F 47 A A F F F 48 A A F F F 49 A A F F F 50 A B F F F 51 A B F F F 52 A B F F F 53 B C F F F 54 A B F F F 55 A A F F F 56 A B F F F 57 A A F F F 58 B C F F F 59 A A F F F 60 A A E E E 61 A A F F F 62 A B F F F 63 A A F F F 64 A A F F F 65 A A F F F 66 A A F F F 67 C C F F F 69 A B F F F 70 B C F F F 71 A B E E F 72 B C F F F 73 C D F F F 74 A B F F F 75 A B F F F 76 D D F F F 77 C C E E F 78 A A F E F 79 A A F E E 80 A A F F F 81 C D E E F 82 C D F E F 83 A A F F F 84 A A F F F 85 A D F E F 86 B C F F F 87 B D F F F 88 D D F F F 89 B B F F F 90 D D F F F 91 C C E F F 92 A A E E F 93 B C F F F 94 A A E E E 95 A A F F F 96 A A D E E 97 A A E E F 98 A A D E F 99 A A E F F 100 A A E F F 101 A A F F F 102 B A F F F 103 A B F F F 104 B C E E F 105 A A F F F

The data in Table 4 clearly indicate that the compounds of the invention have an unexpected and selective anti-proliferation activity against cancer cell growth that is entirely dependent on the class III RTK signaling pathway. Accordingly, these compounds can be used in the treatment of protein kinase-related diseases, such as diseases or disorders mediated by FLT3, PDGFR, c-KIT and/or CSF-1R kinases or mediated by mutant FLT3, PDGFR, c-KIT and/or CSF-1R kinases. A method according to one embodiment of the invention comprises administering to a patient in need thereof an effective amount of a compound of the invention.

EXAMPLE 4: Identification of 6,5-fused dicyclic heteroaryls at position 5 or 6 with position 2 as a nitrogen atom conjugated with heterocyclic pyrazole derivatives (Formula I) demonstrating unexpected and superior specific activity towards FLT3 kinase as well as its mutant.

Formula (I)

As shown in Table 5, Compound 1 and Compound 2 are nearly identical in structure to Examples A through G, respectively, except that the 6,5-fused bicyclic heteroaryl rings in Compound 1 and Compound 2 contain 2 nitrogen atoms at 2 positions, unlike Examples A through G.

Table 5. Comparison of the specific activity in vitro of the inventive compounds of formula (I) with similar compounds

Example Structure Biochemical activity (Average IC ₅₀ , nM) FLT3 FLT3-D835Y Example A 4 6 Example B 64 80 Example C 13 7 Example D 24 17 Example E 27 27 Example F 30 48 Example G 70 106 Connection 1 1.5 0.5 Connection 2 1.0 0.5

Surprisingly, it was observed that Compound 1 and Compound 2 have superior specific activity compared to that of Examples A to G in inhibiting FLT3 kinase, as well as the FLT3-D835Y mutant with approximately 5 or more times better specific activity. Based on the above results, it is surprisingly found that the 6,5-fused bicyclic heteroaryl containing 2 nitrogen atoms at 2 positions exhibits extremely effective inhibitory activity against FLT3 kinase and its mutants.

EXAMPLE 5: Comparison of the specific activity of the compound of formula (I) with the specific activity of structurally related compounds in the limiting part of the claims

The compounds of Examples H to K were found or provided in the preamble of the claims (US 8853207 B2). Compound 40 and Compound 3 were selected to compare their in vitro specific activity with the specific activity of the compounds of Example H. Compound 1 and Compound 2 were selected to compare their in vitro specific activity with the specific activity of Examples I and J. Compound 36 and Compound 30 were selected to compare their in vitro specific activity with the specific activity of Example K.

As shown in Table 6, Compound 40, Compound 3, Compound 1, Compound 2, Compound 36, and Compound 30 are nearly identical in structure to the compounds of Examples H through K, respectively, except that the compounds of the invention have one substituted indazole moiety attached directly to the pyrazole, unlike the limited formula compounds of Examples H through K.

Table 6. Comparison of the specific activity in vitro of compounds possessing the features of the invention with the compounds provided for in the limiting part of the claims

Example Structure Biochemical activity (Average IC ₅₀ , nM) FLT3 FLT3-D835Y Example H
(Ex-78 in US8853207B2) 8 15 Connection 40 0.5 0.4 Connection 3 0.2 0.3 Example I
(Ex-76 in US8853207B2) 12 31 Example J
[Formula (I) in US 8853207 B2) 138 89 Connection 1 1.5 0.5 Connection 2 1.0 0.5 Example K
[Formula (I) in US 8853207 B2) 6 26 Compound 36 0.5 0.4 Connection 30 0,1 0.3

Surprisingly, in contrast to the compounds of the preamble of Examples H to K, the compounds of the invention demonstrated superior specific activity in inhibiting FLT3 kinase and its mutant with approximately 10 or more times greater specific activity.

EXAMPLE 6: Evaluation of the cellular specific activity of compounds of formula (I) in relation to the phosphorylation of FLT3 and its downstream effector STAT5 in the leukemia cell line MV4-11 with constitutive activation of the FLT3 signal.

FLT3 is expressed in AML cells from the majority of patients, and FLT3 is mutated in approximately 30% of AML cells. Mutations include internal tandem duplications (ITDs), present in approximately 25% of AML cells, and tyrosine kinase domain (TKD) point mutations, present in approximately 5% of patients. Both ITDs and AML mutations are activating, causing ligand-independent or constitutive activation of FLT3 receptor signaling, thereby promoting cytokine-independent survival and proliferation of AML cells (Blood 2002;100:1532-42). Both FLT3-ITD and FLT3 TKD mutations result in constitutive activation of FLT3 signaling, which involves the FLT3 kinase pathway and its downstream proliferative signaling pathways, including the Ras/MAPK kinase (MEK)/extracellular signal-regulated kinase (ERK) pathway and the PI3K/Akt pathway. Furthermore, in contrast to wild-type FLT3 signaling, FLT3-ITD efficiently activates the STAT5 pathway. STAT5 induces its target genes, such as cyclin D1, c-myc, and the anti-apoptotic gene p21, which are important for cell growth. These effects may indicate a role for FLT3-ITD in the aberrant growth of leukemic cells. Constitutive activation of FLT3-ITD STAT5 can accelerate AML cell growth (J Hematology/Oncology. 2011; 4: 13.).

The potent compounds of formula (I) in EXAMPLE 2 were evaluated by Western blot analysis for irregular cell specific activity in phosphorylating FLT3 and its downstream effector STAT5 in the leukemia cell line MV4-11 with constitutive activation of FLT3 signaling. For Western blot experiments, MV4-11 cells were cultured in IMDM containing 0.1% FBS overnight. On the day of the assay, three million cells were treated with or without compound for 3 hours in 15 ml tubes. Whole cell lysates were then collected shortly after extraction by adding 2x DBS sample buffers and boiling on a hotplate at 105°C for 15 minutes. The resulting cell lysates were separated by SDS-PAGE and transferred to a PVDF membrane. Protein expression was detected by immunoblotting with the appropriate primary antibody and secondary antibody using standard procedures. Anti-p-FLT3 (Tyr-591), anti-rabbit IgG HRP-coupled secondary, and anti-mouse IgG HRP-coupled secondary were purchased from Cell Signaling Technology. Anti-p-STAT5 (Tyr-694) was purchased from BD biosciences. Anti-actin was purchased from R&D system. Immunoblots were detected by chemiluminescence (SuperSignal ^™ West Femto Maximum Sensitivity Substrate, Thermo) and visualized with the ChemiDoc ^™ MP Imaging System (Bio-Rad). The intensity of the Western blot bands was also quantified using the ChemiDoc ^™ MP imaging system. The relative intensity values of the bands corresponding to the drug treatments at a single concentration (30 nM) were compared with the DMSO control group. The percentage (%) inhibition at each drug concentration was calculated using the formula:

Percentage (%) inhibition = (1 - Relative intensity) × 100.

Selected compounds of formula I showing greater than 50% inhibition of FLT3 phosphorylation and its downstream effector STAT5 upon drug treatment at 30 nM were listed in the following Table 7.

Table 7. Inhibition of phosphorylation of FLT3 and its downstream effector STAT5 in FLT3-ITD-positive MV4-11 leukemia cells

Compound with >50% inhibitory activity on the phosphorylation of FLT3 and its downstream effector STAT5
(at 30 nM treatment) Connection 1, 2, 3, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 28, 29, 30, 31, 32, 33, 34, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 69, 71, 72, 74, 75, 78, 79, 80, 83, 84, 89, 92, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 105.

Some embodiments of the invention relate to methods of treating a disease associated with a protein kinase. A method according to one embodiment of the invention involves administering to a patient in need thereof an effective amount of a compound of the invention.

Although the invention has been described with respect to a limited number of embodiments, those skilled in the art, having the benefit of this disclosure, will appreciate that other embodiments can be devised that do not depart from the scope of the invention as disclosed herein. Accordingly, the scope of the invention should be limited only by the appended claims.

Claims (142)

1. A compound of formula (II), (III), (IV) or (V): Formula (II) Formula (III) Formula (IV) Formula (V) or a stereoisomer, a pharmaceutically acceptable salt, in which G is selected from the group consisting of optionally substituted C6 aryl, optionally substituted 5-6 membered heteroaryl having one or more heteroatoms selected from N and S, optionally substituted 5-8 membered heterocyclyl having one or more heteroatoms selected from O and N, C2-C10 alkynyl and a direct bond; L is selected from the group consisting of -CH=, ^-CHR4- , -( _CH2 ) _q- , ^-NR5- , -O-, -O( _CH2 ) _q- , -C(O)-, -C(O)( _CH2 ) _q- , _-SO2- and a direct bond, wherein each q is individually and independently an integer from 1 to 4; Z is selected from the group consisting of hydrogen, deuterium, halogen, C1-C4 alkyl, C1-C4 alkylamine, C1-C4 dialkylamine, and optionally substituted 4-7 membered heterocyclyl or heterocyclic spiro compound; R ¹ is selected from the group consisting of hydrogen and deuterium; R ² is selected from the group consisting of hydrogen, deuterium, halogen and C1-C6 alkyl; R ³ is selected from the group consisting of hydrogen, deuterium and C1-C6 alkyl; R ⁴ is selected from the group consisting of the group consisting of deuterium and C1-C6 alkyl; and R ⁵ is selected from the group consisting of hydrogen and C1-C6 alkyl. 2. The compound of claim 1, wherein ring G is optionally substituted C6 aryl, optionally substituted 5-6 membered heteroaryl having one or more heteroatoms selected from N and S, optionally substituted 5-8 membered heterocyclyl having one or more heteroatoms selected from O and N, or C2-C10 alkynyl. 3. The compound of claim 2, wherein ring G is selected from the group consisting of: in which R ⁶ is selected from the group consisting of hydrogen, deuterium, halogen, nitro, cyano, trifluoromethyl, trifluoromethoxy, C1-C6 alkyl and C1-C6 alkoxy; each n is individually and independently an integer from 0 to 4; and R ⁷ is hydrogen. 4. A compound according to claim 3, wherein Z is fluoride, chloride, bromide, dimethylamino, or an optionally substituted four-, five-, or six-membered or seven-membered heterocyclyl or heterocyclic spiro compound. 5. The compound of claim 4, wherein Z is selected from the group consisting of: in which R ⁸ is selected from the group consisting of hydrogen, deuterium, halogen, nitro, cyano, trifluoromethyl, trifluoromethoxy, 4-morpholinyl, C1-C6 alkyl and C1-C6 alkoxy; each n is individually and independently an integer from 0 to 4; and R ⁹ is selected from the group consisting of hydrogen, deuterium, and C1-C6 alkyl. 6. The compound according to item 1, which is: 3-(1H-indazol-5-yl)-6-(6-methoxypyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, 3-(1H-indazol-6-yl)-6-(6-methoxypyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, 5-(3-(1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-amine , 6-(6-methoxypyridin-3-yl)-3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, N,N-dimethyl-5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-amine, 4-(4-(3-(1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)benzyl) morpholine, (2S,6R)-4-(4-(3-(1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol- 6-yl)benzyl)-2,6-dimethylmorpholine, 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)morpholine, (2S,6R)-2,6-dimethyl-4-((5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-yl)methyl)morpholine, 4-((5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-yl)methyl)morpholine, 3-(1-methyl-1H-indazol-6-yl)-6-(6-methylpyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, (2S,6R)-2,6-dimethyl-4-(3-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5] cyclopenta[1,2-c]pyrazol-6-yl)benzyl)morpholine, 4-(3-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)morpholine, 3-(1-methyl-1H-indazol-6-yl)-6-(3-((4-methylpiperazin-1-yl)methyl)phenyl)-1,4-dihydrothieno[2',3':4, 5]cyclopenta[1,2-c]pyrazole, (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(4-methylpiperazin-1-yl)methanone, (2R,6S)-4-((5-(3-(1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-yl)methyl)-2,6-dimethylmorpholine, 3-(1-methyl-1H-indazol-6-yl)-6-(2-methylpyridin-4-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, 4-((5-(3-(1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridine -2-yl)methyl)morpholine, ((2S,6R)-2,6-dimethylmorpholino)(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)methanone, 4-(1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole -6-yl)phenyl)ethyl)morpholine, (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(morpholino)methanone, 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenethyl)morpholine, (5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-yl)(morpholino)methanone, 4-(2-methoxy-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazol-6-yl)benzyl)morpholine, 4-(2-fluoro-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazol-6-yl)benzyl)morpholine, 4-(2-methyl-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazol-6-yl)benzyl)morpholine, 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)-2-(trifluoromethyl)benzyl)morpholine, 4-(1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)propyl)morpholine, 4-(3-methoxy-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazol-6-yl)benzyl)morpholine, 3-(1H-indazol-6-yl)-6-(6-methylpyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, (5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-3-yl)(morpholino)methanone, 4-(3-fluoro-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazol-6-yl)benzyl)morpholine, 3-(1-methyl-1H-indazol-6-yl)-6-(6-methylpyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(piperidin-1-yl)methanone, 4-(4-(3-(1-isobutyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)morpholine, 3-(1H-indazol-5-yl)-6-(6-methylpyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, (3-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(morpholino)methanone, (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(pyrrolidin-1-yl)methanone, 3-(1-methyl-1H-indazol-6-yl)-6-(4-(thiomorpholinomethyl)phenyl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c ]pyrazole, 5-(3-(1H-indazol-5-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-amine , (5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyrimidin-2-yl)(morpholino)methanone, 4-(4-(3-(1-isopropyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)morpholine, (2-methoxy-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(morpholino)methanone, (2-fluoro-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)(morpholino)methanone, 5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyrimidin-2-amine, methyl 4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]amrazol-6-yl)-1H-pyrrol-2-yl)(morpholino)methanone, 4-((4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6 -yl)phenyl)sulfonyl)morpholine, (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopental[1,2-c]pyrazol-6-yl)thiophen-2-yl)(morpholino)methanone, N,N-dimethyl-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)benzamide, N,N-dimethyl-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)benzenesulfonamide, 4-((5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)thiophen-2-yl)methyl)morpholine, 4-((4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)thiophen-2-yl)methyl)morpholine, 4-(2,6-difluoro-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2- c]pyrazol-6-yl)benzyl)morpholine, (2S,6R)-2,6-dimethyl-4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5] cyclopenta[1,2-c]pyrazol-6-yl)benzyl)morpholine, 4-((6-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-3-yl)methyl)morpholine, (2-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-4-yl)(morpholino)methanone, (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopental[1,2-c]pyrazol-6-yl)-3,6-dihydropyridin-1(2H)-yl)(morpholino)methanone, tert-butyl 4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)piperidine-1-carboxylate, 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)morpholin-3-one, (4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)thiazol-2-yl)(morpholino)methanone, 1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)piperidin-2-one, 3-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)oxazolidin-2-one, 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)piperazin-2-one, 1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)pyrrolidin-2-one, 1,3-dimethyl-4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2- c]pyrazol-6-yl)benzyl)piperazin-2-one, 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenethyl)morpholin-3-one, 6-(4-((3,3-difluoropyrrolidin-1-yl)methyl)phenyl)-3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, 4-(4-(3-(3-bromo-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)morpholine, 4-(2-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole -6-yl)phenoxy)ethyl)morpholine, 4-(2-(3-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole -6-yl)phenoxy)ethyl)morpholine, 4-(3-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)prop-2-yn-1-yl)morpholine, N-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)tetrahydro-2H-pyran-4-amine, N-methyl-N-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)tetrahydro-2H-pyran-4-amine, (R)-3-methyl-4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1, 2-c]pyrazol-6-yl)benzyl)morpholine, (S)-3-methyl-4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1, 2-c]pyrazol-6-yl)benzyl)morpholine, 3-(1-methyl-1H-indazol-6-yl)-6-(4-((tetrahydro-4H-pyran-4-ylidene)methyl)phenyl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, 3-(1-methyl-1H-indazol-6-yl)-6-(4-((tetrahydro-2H-pyran-4-yl)methyl)phenyl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, 4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)morpholine, 4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)morpholin-3-one, 4-(1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole -6-yl)benzyl)piperidin-4-yl)morpholine, 4-(4-(3-(3-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)morpholine, 4-(5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)pyridin-2-yl)morpholine, 3-(1H-indazol-6-yl)-6-(4-((4-methylpiperazin-1-yl)methyl)phenyl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, 3-(1-methyl-1H-indazol-6-yl)-6-(4-((4-methylpiperazin-1-yl)methyl)phenyl)-1,4-dihydrothieno[2',3':4, 5]cyclopenta[1,2-c]pyrazole, 3-(1-methyl-1H-indazol-6-yl)-6-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, 6-(2-chloropyridin-4-yl)-3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, 3-(1-methyl-1H-indazol-6-yl)-6-(2-(4-methylpiperazin-1-yl)pyridin-4-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, 4-(2,3-difluoro-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2- c]pyrazol-6-yl)benzyl)morpholine, 4-(4-(3-(1-methyl-1H-benzo[d][1,2,3]triazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta [1,2-c]pyrazol-6-yl)benzyl)morpholine, 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)-3-(trifluoromethoxy)benzyl)morpholine, 4-(3-methyl-4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazol-6-yl)benzyl)morpholine, 5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-amine, N,N-dimethyl-1-(5-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-2-yl)methanamine, N,N-dimethyl-1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)phenyl)methanamine, 4-((6-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6-yl)pyridin-3-yl)methyl)morpholine, 3-(1-methyl-1H-indazol-6-yl)-6-(piperidin-4-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c] pyrazole, 4-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)-1,4-oxazepane, 1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)piperedin-1-yl)-2-morpholinoethan-1-one, 3-(1-methyl-1H-indazol-6-yl)-6-(4-(pyrrolidin-1-ylmethyl)phenyl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, 3-(1-methyl-1H-indazol-6-yl)-6-(4-(piperidin-1-ylmethyl)phenyl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazole, 1-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)piperazin-2-one, 6-(4-(3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1,2-c]pyrazol-6- yl)benzyl)-2-oxa-6-azaspiro[3.3]heptane, 6-(4-(azetidin-1-ylmethyl)phenyl)-3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4,5]cyclopenta[1 ,2-c]pyrazole, 6-(4-((3,3-difluoroazetidin-1-yl)methyl)phenyl)-3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3': 4,5]cyclopenta[1,2-c]pyrazole, or 6-(4-((3-methoxyazetidin-1-yl)methyl)phenyl)-3-(1-methyl-1H-indazol-6-yl)-1,4-dihydrothieno[2',3':4, 5]cyclopenta[1,2-c]pyrazole. 7. A pharmaceutical composition for the treatment of a disease or disorder mediated by FLT3 and/or PDGFR kinases or mediated by a mutant FTL3 kinase and/or PDGFR kinases, comprising a compound according to any one of claims 1-6, its stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable diluent or carrier. 8. A method for treating a disease or disorder mediated by FLT3 kinases and/or PDGFR kinases or mediated by a mutant kinase of FLT3 kinases and/or PDGFR kinases, comprising: administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound according to any one of claims 1-6, a stereoisomer thereof or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable diluent or carrier or protein targeting module. 9. The method of claim 8, wherein the disease or disorder is cancer. 10. The method of claim 9, wherein the cancer is selected from the group consisting of multiple myeloma, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), prostate cancer, breast cancer, ovarian cancer, melanoma, glioblastoma multiforme, giant cell tumor of bone, non-small cell lung cancer, giant cell tumor of the tendon sheath, tumor metastasis to other tissues, myelofibrosis, pigmented villonodular synovitis, or gastrointestinal stromal tumor.