Inhibitors of rock2

Abstract

The present disclosure relates to inhibitors of Rho-associated coiled coil containing protein kinase (ROCK), pharmaceutical compositions comprising the same, and use thereof for the prevention or treatment of a disease mediated by the ROCK. Particularly, the inhibitors of ROCK are selective for the inhibition of ROCK2.

Description

322930.48876INHIBITORS OF ROCK2FIELD

[0001] The present disclosure relates to inhibitors of Rho-associated coiled-coil containing protein kinase 2 (ROCK2), pharmaceutical compositions comprising the same, and uses thereof for the prevention or treatment of diseases mediated by the ROCK2.BACKGROUND

[0002] Rho-associated coiled-coil containing protein kinase (ROCK) is a serine / threonine kinase from the AGC (PKA, PKG, and PKC) kinase family and comprises two isoforms, ROCK1 and ROCK2. The two isoforms are expressed and regulated differently in specific tissues. For example, ROCK1 is ubiquitously expressed at a relatively high level, while ROCK2 is preferentially expressed in certain tissues including heart, brain and skeletal muscle. ROCK is a target of the small GTPase Rho and is involved in diverse cellular activities achieved by phosphorylating downstream effector proteins (MLC, LIMK, ERM, MARCKS, CRMP-2, etc.). Studies have shown that various diseases (e.g., pulmonary fibrosis, cardiac-cerebral vascular disease, neurological disease, cancer, etc.) are related to the pathways mediated by ROCK. As such, ROCK has been considered as an important target in the development of novel drugs.SUMMARY

[0003] In one aspect, the disclosure provides inhibitors of ROCK2 having the Formula I:or a pharmaceutically acceptable salt thereof, wherein:X1is selected from the group consisting of CH2, O and NR4;X2is selected from the group consisting of CH2, O and NR4;wherein one of X1or X2is CH2; and one of X1or X2is O or NR4;R1is selected from the group consisting of H, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3- C7 cycloalkyl, C5-C10 aryl, C5-C10 heteroaryl, 3- to 10-membered heterocyclyl, -Ci-Ce alkyl-(C3-C? cycloalkyl), -Ci-Ce alkyl-(C5-Cio aryl), -Ci-Ce alkyl-(C5-Cio heteroaryl), - Ci-Ce alkyl-(3- to 10-membered heterocyclyl), -(Ci-Ce alkyl)-NRnR12, -(Ci-Ce alkyl)- OR11, and -C(=O)-N(R11)(R12), and wherein each alkyl, alkenyl, cycloalkyl, aryl, heteroaryl and heterocyclyl may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;each R11and R12are independently selected from the group consisting of H and Ci-Ce alkyl;or alternatively, R11and R12are taken together when both are attached to the same nitrogen to form a 4- to 7- membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from the group consisting of N, O and S, and which heterocyclic ring is unsubstituted or is substituted with 1 to 3 substituents selected from the group consisting of halo, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, -CN, -NH2, C1-C3 perfluoroalkyl, -OH, -O-(Ci-Ce alkyl), and -(Ci-Ce alkyl)-OH;R2and R3are each independently selected from the group consisting of H, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, 3- to 10-membered heterocyclyl, Ce-Cio aryl, 5- to 14-membered heteroaryl, wherein each alkyl, alkenyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;alternatively R2and R3are taken together with the nitrogen to which they are attached to provide (i) a 4- to 6-membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from N, O and S, or (ii) a 5- to 10-membered hetero bicyclic ring system having from 0 to 3 additional ring heteroatoms selected from N, O and S; wherein the heterocyclic ring or the bicyclic ring system are unsubstituted or are substituted with from 1 to 4 substituents selected from the group consisting of halo, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, -CN, Ci-Ce fluoroalkyl, C1-C3 perfluoro alkyl, - OR11, oxo, and -NH2; andR4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

[0004] In another aspect, the disclosure provides method for the treatment of a disease or disorder mediated by ROCK2, wherein the method comprises administering to a subject in 2182500867.1need thereof an effective amount of ROCK 2 inhibitor provided herein or a pharmaceutically acceptable salt thereof.

[0005] In embodiments provided in this disclosure, the methods are used to treat a disease or disorder selected from the group consisting of fibrotic diseases, inflammatory diseases, autoimmune diseases, cardiovascular disorders, central nervous system disorders, neoplastic diseases, metabolic syndromes, ocular diseases, renal diseases, pulmonary diseases, muscular dystrophy, sickle cell disease, and viral diseases.BRIEF DESCRIPTION OF THE FIGURES

[0006] FIG. 1 shows the tracer optimization curves of ROCK 1 and ROCK2 expressed in HEK293 cells by NanoBRET®.

[0007] FIG. 2 shows the dose response curves of for the compound of Example 1 in ROCK1 and ROCK2 expressed in HEK293 cells by NanoBRET®.DETAILED DESCRIPTION

[0008] The present invention will now be further described. In the following passages, different aspects of the invention are provided. Each aspect presented may be combined with any other aspect or aspects unless clearly indicated to the contrary. In particular, any feature indicated as being preferred or advantageous may be combined with any other feature or features indicated as being preferred or advantageous.

[0009] The compounds, compositions and methods described herein provide selective inhibitors of Rho-associated coiled-coil kinase 2 (ROCK2) for use in the treatment of diseases or disorders, including fibrotic diseases, inflammatory diseases, autoimmune diseases, cardiovascular disorders, central nervous system disorders, neoplastic diseases, metabolic syndromes, ocular diseases, renal diseases, pulmonary diseases, muscular dystrophy, sickle cell disease, and viral diseases.

[0010] The compounds for use in the methods and compositions disclosed herein are ROCK inhibitors, and in particular ROCK2 selective inhibitors. The compounds provide excellent inhibitory activity of ROCK (preferably ROCK2) and good selectivity (higher selectivity towards ROCK2 as compared with ROCK1). The compounds may additionally provide one or more of good physicochemical properties (e.g., solubility, physical and / or chemical stability), improved pharmacokinetic properties (e.g., improved bioavailability, proper half-life and duration of action), improved penetrance of the blood-brain barrier, and improved safety (low toxicity and / or less side effects, wide therapeutic window).3182500867.1

[0011] In one aspect, the disclosure provides inhibitors of ROCK2 having the Formula I:or a pharmaceutically acceptable salt thereof, wherein:X1is selected from the group consisting of CH2, O and NR4;X2is selected from the group consisting of CH2, O and NR4;wherein one of X1or X2is CH2; and one of X1or X2is O or NR4;R1is selected from the group consisting of H, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3- C7 cycloalkyl, C5-C10 aryl, C5-C10 heteroaryl, 3- to 10-membered heterocyclyl, -Ci-Ce alkyl-(C3-C? cycloalkyl), -Ci-Ce alkyl-(C5-Cio aryl), -Ci-Ce alkyl-(C5-Cio heteroaryl), - Ci-Ce alkyl-(3- to 10-membered heterocyclyl), -(Ci-Ce alkyl)-NRnR12, -(Ci-Ce alkyl)- OR11, and -C(=O)-N(R11)(R12), and wherein each alkyl, alkenyl, cycloalkyl, aryl, heteroaryl and heterocyclyl may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;each R11and R12are independently selected from the group consisting of H and Ci-Ce alkyl;or alternatively, R11and R12are taken together when both are attached to the same nitrogen to form a 4- to 7- membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from the group consisting of N, O and S, and which heterocyclic ring is unsubstituted or is substituted with 1 to 3 substituents selected from the group consisting of halo, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, -CN, -NH2, C1-C3 perfluoroalkyl, -OH, -O-(Ci-Ce alkyl), and -(Ci-Ce alkyl)-OH;R2and R3are each independently selected from the group consisting of H, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, 3- to 10-membered heterocyclyl, Ce-Cio aryl, 5- to 14-membered heteroaryl, wherein each alkyl, alkenyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;4182500867.1alternatively R2and R3are taken together with the nitrogen to which they are attached to provide (i) a 4- to 6-membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from N, O and S, or (ii) a 5- to 10-membered hetero bicyclic ring system having from 0 to 3 additional ring heteroatoms selected from N, O and S; wherein the heterocyclic ring or the bicyclic ring system are unsubstituted or are substituted with from 1 to 4 substituents selected from the group consisting of halo, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, -CN, Ci-Ce fluoroalkyl, C1-C3 perfluoro alkyl, - OR11, oxo, and -NH2; andR4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

[0012] In embodiments of the above compounds, R4is selected from hydrogen, C1-C3 alkyl and C3-C5 cycloalkyl, wherein the C1-C3 alkyl and C3-C5 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy. In some particular embodiments, R4is selected from H, methyl, ethyl, and cyclopropyl, which may be optionally substituted by 1 to 3 substituents selected from F, Cl and -OCH3.

[0013] In embodiments of the above compounds, R2and R3may be taken together with the nitrogen to which they are attached to provide a 4- or 5-membered heterocycle, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of Ci-Ce alkyl, Ci-Ce-fluoroalkyl and halo, and particularly fluoro.

[0014] In embodiments of the above compounds, R1is selected from H and Ci-Ce alkyl, and particularly R1may be methyl.

[0015] In embodiments, the disclosure provides inhibitors of ROCK2 having the Formula IA:or a pharmaceutically acceptable salt thereof, wherein:X1is selected from the group consisting of CH2, O and NR4;X2is selected from the group consisting of CH2, O and NR4;5182500867.1wherein one of X1or X2is CH2; and one of X1or X2is O or NR4;R1Ais selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;R2Aand R3Aare each independently selected from the group consisting of H, Ci-Ce alkyl, C2- Ce alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, wherein each alkyl, alkenyl, cycloalkyl, may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;alternatively R2Aand R3Aare taken together with the nitrogen to which they are attached to provide a 4- to 6-membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from N, O and S, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of halo, C1-C3 alkyl, -CN, C1-C3 fluoroalkyl, C1-C3 perfluoro alkyl, -NH2, -OH, and -O-C1-C3 alkyl; andR4is selected from the group consisting of H, Ci-Ce alkyl and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

[0016] In embodiments of the above compounds, R4is selected from hydrogen, C1-C3 alkyl and C3-C5 cycloalkyl, wherein the C1-C3 alkyl and C3-C5 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy. In some particular embodiments, R4is selected from H, methyl, ethyl, and cyclopropyl, which may be optionally substituted by 1 to 3 substituents selected from F, Cl and -OCH3.

[0017] In embodiments of the above compounds, R2Aand R3Amay be taken together with the nitrogen to which they are attached to provide a 4- or 5-membered heterocycle, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of Ci-Ce alkyl, Ci-Ce-fluoroalkyl and halo, and particularly fluoro.

[0018] In embodiments of the above compounds, R1Ais selected from H and Ci-Ce alkyl, and particularly R1Amay be methyl.

[0019] In embodiments, the disclosure provides inhibitors of ROCK2 having the Formula IB:6182500867.1or a pharmaceutically acceptable salt thereof, wherein:X1is selected from the group consisting of CH2, O and NR4;X2is selected from the group consisting of CH2, O and NR4;wherein one of X1or X2is CH2; and one of X1or X2is O or NR4;R1Bis selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;each R5is independently selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;n is 1 or 2;m is 0, 1 or 2; andR4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

[0020] In embodiments of the above compounds, R4is selected from hydrogen, C1-C3 alkyl and C3-C5 cycloalkyl, wherein the C1-C3 alkyl and C3-C5 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy. In some particular embodiments, R4is selected from H, methyl, ethyl, and cyclopropyl, which may be optionally substituted by 1 to 3 substituents selected from F, Cl and -OCH3.

[0021] In embodiments of the above compounds, R1Bis selected from H and Ci-Ce alkyl, and particularly R1Bmay be methyl.

[0022] In embodiments of the above compounds, m may be 0. Alternatively, m may be 1 or 2. Each R5may be selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, Cl and F, and particularly, R5is F.

[0023] In one aspect, the disclosure provides inhibitors of ROCK2 having the Formula IIA:7182500867.1or a pharmaceutically acceptable salt thereof, wherein:X1is selected from the group consisting of O and NR4;R1Ais selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;R2Aand R3Aare each independently selected from the group consisting of H, Ci-Ce alkyl, C2- Ce alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, wherein each alkyl, alkenyl, cycloalkyl, may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;alternatively R2Aand R3Aare taken together with the nitrogen to which they are attached to provide a 4- to 6-membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from N, O and S, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of halo, C1-C3 alkyl, -CN, C1-C3 fluoroalkyl, C1-C3 perfluoro alkyl, -NH2, -OH, and -O-C1-C3 alkyl; and R4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

[0024] In embodiments of the above compounds, R4is selected from hydrogen, C1-C3 alkyl and C3-C5 cycloalkyl, wherein the C1-C3 alkyl and C3-C5 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy. In some particular embodiments, R4is selected from H, methyl, ethyl, and cyclopropyl, which may be optionally substituted by 1 to 3 substituents selected from F, Cl and -OCH3.

[0025] In embodiments of the above compounds, R2Aand R3Amay be taken together with the nitrogen to which they are attached to provide a 4- or 5-membered heterocycle, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of Ci-Ce alkyl, Ci-Ce-fluoroalkyl and halo, and particularly fluoro.

[0026] In embodiments of the above compounds, R1Ais selected from H and Ci-Ce alkyl, and particularly R1Amay be methyl.8182500867.1

[0027] In some embodiments, this disclosure provides inhibitors of ROCK2 having the Formula IIIA:R3A / IIIA or a pharmaceutically acceptable salt thereof, wherein:R1Ais selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;R2Aand R3Aare each independently selected from the group consisting of H, Ci-Ce alkyl, C2- Ce alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, wherein each alkyl, alkenyl, cycloalkyl, may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2; andalternatively R2Aand R3Aare taken together with the nitrogen to which they are attached to provide a 4- to 6-membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from N, O and S, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of halo, C1-C3 alkyl, -CN, C1-C3 fluoroalkyl, C1-C3 perfluoro alkyl, -NH2, -OH, and -O-C1-C3 alkyl.

[0028] In embodiments of the above compounds, R2Aand R3Amay be taken together with the nitrogen to which they are attached to provide a 4- or 5-membered heterocycle, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of Ci-Ce alkyl, Ci-Ce-fluoroalkyl and halo, and particularly fluoro.

[0029] In embodiments of the above compounds, R1Ais selected from H and Ci-Ce alkyl, and particularly R1Amay be methyl.

[0030] In some embodiments, this disclosure provides inhibitors of ROCK2 having the Formula IIIB:182500867.1or a pharmaceutically acceptable salt thereof, wherein:R1Bis selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;each R5is independently selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;n is 1 or 2; andm is 0, 1 or 2.

[0031] In embodiments of the above compounds, R1Bis selected from H and Ci-Ce alkyl, and particularly R1Bmay be methyl.

[0032] In embodiments of the above compounds, m may be 0. Alternatively, m may be 1 or 2. Each R5may be selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, Cl and F, and particularly, R5is F.

[0033] In some embodiments, this disclosure provides inhibitors of ROCK2 having Formula IVA:or a pharmaceutically acceptable salt thereof, wherein:R1Ais selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;R2Aand R3Aare each independently selected from the group consisting of H, Ci-Ce alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C7 cycloalkyl, wherein each alkyl, alkenyl, cycloalkyl, may 10182500867.1optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;alternatively R2Aand R3Aare taken together with the nitrogen to which they are attached to provide a 4- to 6-membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from N, O and S, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of halo, C1-C3 alkyl, -CN, C1-C3 fluoroalkyl, C1-C3 perfluoro alkyl, -NH2, -OH, and -O-C1-C3 alkyl; andR4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

[0034] In embodiments of the above compounds, R4is selected from hydrogen, C1-C3 alkyl and C3-C5 cycloalkyl, wherein the C1-C3 alkyl and C3-C5 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy. In some particular embodiments, R4is selected from H, methyl, ethyl, and cyclopropyl, which may be optionally substituted by 1 to 3 substituents selected from F, Cl and -OCH3.

[0035] In embodiments of the above compounds, R2Aand R3Amay be taken together with the nitrogen to which they are attached to provide a 4- or 5-membered heterocycle, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of Ci-Ce alkyl, Ci-Ce-fluoroalkyl and halo, and particularly fluoro.

[0036] In embodiments of the above compounds, R1Ais selected from H and Ci-Ce alkyl, and particularly R1Amay be methyl.

[0037] In some embodiments, this disclosure provides inhibitors of ROCK2 having Formula IVB:or a pharmaceutically acceptable salt thereof, wherein:11182500867.1R1Bis selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;each R5is independently selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;n is 1 or 2;m is 0, 1 or 2; andR4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

[0038] In embodiments of the above compounds, R1Bis selected from H and Ci-Ce alkyl, and particularly R1Bmay be methyl.

[0039] In embodiments of the above compounds, m may be 0. Alternatively, m may be 1 or 2. Each R5may be selected from C1-C3 alkyl, C1-C3 perfluoroalkyl Cl and F, and particularly, R5is F.

[0040] In some embodiments, this disclosure provides inhibitors of ROCK2 having Formula VA:or a pharmaceutically acceptable salt thereof, wherein:R1Ais selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;R2Aand R3Aare each independently selected from the group consisting of H, Ci-Ce alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C7 cycloalkyl, wherein each alkyl, alkenyl, cycloalkyl, may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;alternatively R2Aand R3Aare taken together with the nitrogen to which they are attached to provide a 4- to 6-membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from N, O and S, wherein the heterocyclic ring is unsubstituted or12182500867.1is substituted with from 1 to 4 substituents selected from the group consisting of halo, C1-C3 alkyl, -CN, C1-C3 fluoroalkyl, C1-C3 perfluoro alkyl, -NH2, -OH, and -O-C1-C3 alkyl; andR4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

[0041] In embodiments of the above compounds, R4is selected from hydrogen, C1-C3 alkyl and C3-C5 cycloalkyl, wherein the C1-C3 alkyl and C3-C5 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy. In some particular embodiments, R4is selected from H, methyl, ethyl, and cyclopropyl, which may be optionally substituted by 1 to 3 substituents selected from F, Cl and -OCH3.

[0042] In embodiments of the above compounds, R2Aand R3Amay be taken together with the nitrogen to which they are attached to provide a 4- or 5-membered heterocycle, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of Ci-Ce alkyl, Ci-Ce-fluoroalkyl and halo, and particularly fluoro.

[0043] In embodiments of the above compounds, R1Ais selected from H and Ci-Ce alkyl, and particularly R1Amay be methyl.

[0044] In some embodiments, this disclosure provides inhibitors of ROCK2 having Formula VB:or a pharmaceutically acceptable salt thereof, wherein:R1Bis selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;each R5is selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2; n is 1 or 2;m is 0, 1 or 2; and13182500867.1R4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

[0045] In embodiments of the above compounds, R1Bis selected from H and Ci-Ce alkyl, and particularly R1Bmay be methyl.

[0046] In embodiments of the above compounds, m may be 0. Alternatively, m may be 1 or 2. Each R5may be selected from C1-C3 alkyl, C1-C3 perfluoroalkyl Cl and F, and particularly, R5is F.

[0047] In embodiments, the inhibitor of ROCK2 is selected from compounds having the formula:182500867.1or a pharmaceutically acceptable salt thereof.

[0048] In an embodiment, this disclosure provides an inhibitor of ROCK2 which is Compound 1:or a pharmaceutically acceptable salt thereof. Compound 1 has the chemical name (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)-7,8-dihydro-5H-pyrano[4,3-d]pyrimidin-2-yl)-1-methyl-1H-indol-2-yl)(3,3-difluoroazetidin-1-yl)methanone and may optionally in the form of a pharmaceutically acceptable salt.182500867.1

[0049] The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0050] The term "halogen" or "halo" designates -F, -Cl, -Br or -I. Preferred halogens are -F, -Cl and -Br.

[0051] The term "hydroxyl" means -OH.

[0052] The term “oxo” as used herein refers to an oxygen atom that has a double bond to another atom (i.e., the substituent =0), particularly to carbon.

[0053] The term "alkyl" refers to the radical of saturated aliphatic groups, including straight-chain alkyl groups and branched-chain alkyl groups. Accordingly, Ci-Ce alkyl includes methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, and the like. Fluoroalkyl refers to an alkyl group in which one or more of the hydrogens has been replaced by -F. Perfluoroalkyl refers to an alkyl group in which each of the hydrogens has been replaced by -F.

[0054] The term “cycloalkyl” refers to saturated, carbocyclic groups having from 3 to 7 carbons in the ring. Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0055] The term “alkenyl” refers to a linear or branched hydrocarbyl having a double bond and 2-6 carbon atoms (“C2-C6 alkenyl”). The alkenyl includes vinyl, 1 -propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3 -pentenyl, 4-pentenyl, 2-h exenyl, 3 -hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl, 4-methyl-3 -pentenyl, and the like. When the compound of the present disclosure contains an alkenyl group, the compound may exist as the E-form, the Z-form, or any mixture thereof.

[0056] The term “alkynyl” refers to a linear or branched hydrocarbyl having a triple bond and 2-6 carbon atoms (“C2-C6 alkynyl). The alkynyl includes ethynyl, propynyl, and the like.

[0057] The term "aryl" as used herein includes 5- and 6-membered single-ring aromatic groups that may include from zero to four heteroatoms, for example, benzene, pyrene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like. Those aryl groups having heteroatoms in the ring structure may also be referred to as "aryl heterocycles", "heteroaromatics" or "heteroaryl". The term “aryl” also includes 7- to 14-membered polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings (the rings are "fused rings") wherein at least one of the rings is aromatic (including heteroaryl), e.g., the other cyclic rings can be fused cycloalkyls, cycloalkenyls, aryls, heteroaryl and / or heterocyclic groups. Single-ring heteroaryl groups may have from 1 to 3 ring heteroatoms 16182500867.1and fused polycyclic heteroaryl groups may have from 1 to 5 ring heteroatoms, wherein the ring heteroatoms are selected from N, O and S.

[0058] The terms "heterocyclyl" or "heterocyclic group" refer to 3- to 10-membered ring structures, more preferably 5- or 6-membered rings, whose ring structures include one to four heteroatoms. Heterocycles can also be polycycles. Heterocyclic groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxathiin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenarsazine, phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane, thiolane, oxazole, piperidine, piperazine, morpholine, lactones, lactams such as azetidinones and pyrrolidinones, sultams, sultones, and the like.

[0059] The term “aralkyl”, as used herein, refers to a Ci-Ce alkyl group substituted with an aryl group (e.g., an aromatic or heteroaromatic group).

[0060] As used herein, the definition of each expression, e.g. alkyl, m, n, R, etc., when it occurs more than once in any structure, is intended to be independent of its definition elsewhere in the same structure.

[0061] It will be understood that "substituted", "substitution" or "substituted with" includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc.

[0062] Certain compounds provided in this disclosure may exist in particular geometric or stereoisomeric forms. The disclosure contemplates all such compounds, including cis- and trans-isomers, R- and 5-enantiomers, diastereomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention. Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are included in this invention.

[0063] The compounds disclosed herein may exist in particular tautomeric forms, in which the forms are isomers which may be interchanged by migration of one or more hydrogens (protons) and one or more double bonds. Each depiction herein of a particular tautomeric form is intended to include each other tautomeric form.17182500867.1

[0064] The compounds disclosed herein may have one or more deuterium atoms replacing hydrogen atom(s) at any position.

[0065] The term "pharmaceutically-acceptable salts" refers to the relatively non-toxic, inorganic and organic acid addition salts of compounds disclosed herein and inorganic and organic basic addition salts of the compounds disclosed herein.

[0066] As set out above, certain embodiments of the ROCK2 inhibitors may contain a basic functional group, such as amino, and are capable of forming pharmaceutically-acceptable salts with pharmaceutically-acceptable acids. These salts can be prepared in situ in the administration vehicle or the dosage form manufacturing process, or by separately reacting a purified compound of the invention in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed during subsequent purification.Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts and the like. (See, for example, Berge et al. (1977) " Pharmaceutical Salts", J. Pharm. Sci. 66:1-19).

[0067] The pharmaceutically acceptable salts of the subject compounds include the conventional nontoxic salts or quaternary ammonium salts of the compounds, e.g., from nontoxic organic or inorganic acids. For example, such conventional nontoxic salts include those derived from inorganic acids such as hydrochloride, hydrobromic, sulfuric, sulfamic, phosphoric, nitric, and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicyclic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isothionic, and the like.

[0068] In other cases, the compounds provided in this disclosure may contain one or more acidic functional groups and, thus, are capable of forming pharmaceutically-acceptable salts with pharmaceutically-acceptable bases. These salts can likewise be prepared in situ in the administration vehicle or the dosage form manufacturing process, or by separately reacting the purified compound in its free acid form with a suitable base, such as the hydroxide, carbonate or bicarbonate of a pharmaceutically-acceptable metal cation, with ammonia, or with a pharmaceutically-acceptable organic primary, secondary or tertiary amine.Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts and the like. Representative organic amines useful for the18182500867.1formation of base addition salts include ethylamine, di ethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and the like. (See, for example, Berge et al., supra).Method of Treatment

[0069] The present disclosure provides a method for the prevention or treatment of a disease mediated by ROCK2, wherein the method comprises administering to a subject in need thereof an effective amount of a ROCK2 inhibitor as disclosed herein or a pharmaceutically acceptable salt thereof.

[0070] In some embodiments, the present disclosure provides methods for the treatment of at least one disease or disorder selected from the group comprising fibrotic diseases, inflammatory diseases, and autoimmune diseases, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound or a composition as defined herein.

[0071] In other embodiments, the disclosure provides methods for the treatment of a cardiovascular disorder, a central nervous system disorder, a neoplastic disease, or a metabolic syndrome, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound or a composition as defined herein.

[0072] In some embodiments, the disease mediated by ROCK2 is an autoimmune disorder including rheumatoid arthritis, systemic lupus erythematosus (SLE; lupus), psoriasis, psoriatic arthritis, multiple sclerosis, Crohn’s disease, ulcerative colitis, atopic dermatitis, eczema, or graft-versus-host disease (GVHD; acute and chronic), idiopathic pulmonary fibrosis and scleroderma.

[0073] Other autoimmune disorders that may be treated according to the methods provided in this disclosure include acute disseminated encephalomyelitis (ADEM), acute necrotizing hemorrhagic leukoencephalitis, Addison’s disease, alopecia areata, amyloidosis, ankylosing spondylitis, anti-GBM / anti-TBM nephritis, antiphospholipid syndrome (APS), autoimmune angioedema, autoimmune aplastic anemia, autoimmune dysautonomia, autoimmune hepatitis, autoimmune hyperlipidemia, autoimmune immunodeficiency, autoimmune inner ear disease (AIED), autoimmune myocarditis, autoimmune oophoritis, autoimmune pancreatitis, autoimmune retinopathy, autoimmune thyroid disease, autoimmune urticaria, axonal & neuronal neuropathies, Balo disease, Behcet’s disease, bullous pemphigoid, cardiomyopathy, Castleman disease, Celiac disease, Chagas disease, chronic inflammatory demyelinating polyneuropathy (CIDP), chronic recurrent multifocal ostomy elitis (CRMO), Churg-Strauss syndrome, Cogan’s syndrome, coxsackie myocarditis,19182500867.1CREST disease, demyelinating neuropathies, dermatitis herpetiformis, dermatomyositis, Devic’s disease (neuromyelitis optica), discoid lupus, Dressier’s syndrome, eosinophilic esophagitis, eosinophilic fasciitis, erythema nodosum, Evans syndrome, fibrosing alveolitis, giant cell arteritis (temporal arteritis), giant cell myocarditis, glomerulonephritis, Goodpasture’s syndrome, granulomatosis with polyangiitis (GPA), Graves’ disease, Guillain-Barre syndrome, Hashimoto’s encephalitis, Hashimoto’s thyroiditis, hemolytic anemia, Henoch-Schonlein purpura, Hypogammaglobulinemia, idiopathic thrombocytopenic purpura (ITP), IgA nephropathy, IgG4-related sclerosing disease, immunoregulatory lipoproteins, inclusion body myositis, interstitial cystitis, juvenile arthritis, juvenile myositis, Kawasaki syndrome, Lambert-Eaton syndrome, Leukocytoclastic vasculitis, lichen planus, lichen sclerosus, ligneous conjunctivitis, Linear IgA disease (LAD), Meniere’s disease, microscopic polyangiitis, mixed connective tissue disease (MCTD), Mooren’s ulcer, Mucha-Habermann disease, myasthenia gravis, myositis, neuromyelitis optica, neutropenia, ocular cicatricial pemphigoid, optic neuritis, palindromic rheumatism, pediatric autoimmune neuropsychiatric disorders associated with streptococcus (PANDAS), paraneoplastic cerebellar degeneration, paroxysmal nocturnal hemoglobinuria (PNH), Parry Romberg syndrome, Parsonnage-Turner syndrome, pars planitis (peripheral uveitis), pemphigus, peripheral neuropathy, perivenous encephalomyelitis, pernicious anemia, POEMS syndrome, polyarteritis nodosa, autoimmune polyglandular syndromes (type I, II, III), polymyalgia rheumatica, polymyositis, post myocardial infarction syndrome, post pericardiotomy syndrome, progesterone dermatitis, primary biliary cirrhosis, primary sclerosing cholangitis, psoriatic arthritis, pyoderma gangrenosum, pure red cell aplasia, Raynaud’s phenomenon, reactive arthritis, reflex sympathetic dystrophy, Reiter’s syndrome, relapsing polychondritis, retroperitoneal fibrosis, sarcoidosis, Schmidt syndrome, scleritis, Sjogren’s syndrome, Sperm & testicular autoimmunity, stiff person syndrome, subacute bacterial endocarditis (SBE), Susac’s syndrome, sympathetic ophthalmia, Takayasu’s arteritis, temporal arteritis / Giant cell arteritis, thrombocytopenic purpura (TTP), Tolosa-Hunt syndrome, Transverse myelitis, undifferentiated connective tissue disease (UCTD), type-1 autoimmune diabetes, uveitis, vasculitis, vesiculobullous dermatosis, vitiligo, and Wegener’s granulomatosis (granulomatosis with polyangiitis; GPA).

[0074] Inflammatory disorders that can be treated by methods provided in this disclosure include, but are not limited to, cardiovascular inflammation, pulmonary inflammation, renal inflammation, arteriosclerosis and sepsis.20182500867.1

[0075] Fibrotic disorders that can be treated by methods provided in this disclosure include idiopathic pulmonary fibrosis, renal fibrosis, kidney fibrosis, ocular fibrosis, cardiac fibrosis, NASH, scleroderma, systemic sclerosis, and cirrhosis.

[0076] In another embodiment, the disclosure provides a method for the treatment of muscular dystrophy (Duchenne muscular dystrophy). In another embodiment, the disclosure provides a method for the treatment of myotonic dystrophy.

[0077] In other embodiments, the ROCK2 inhibitors provided herein may be used to inhibit tumor cell growth and metastasis, and angiogenesis, and are useful for treating neoplastic diseases. Neoplastic diseases include any malignant growth or tumor caused by abnormal or uncontrolled cell division. Neoplastic diseases include lymphoma, carcinoma, leukemia, sarcoma and blastoma. Non-limiting examples include squamous cell cancer, small-cell lung cancer, pituitary cancer, esophageal cancer, astrocytoma, soft tissue sarcoma, non-small cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, bladder cancer, liver cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, brain cancer, endometrial cancer, testis cancer, cholangiocarcinoma, gallbladder carcinoma, gastric cancer, melanoma, and various types of head and neck cancer.

[0078] In other embodiments, the ROCK2 inhibitors provided herein may be used in the treatment of a cardiovascular disorder including hypertension, cardiomyopathy, cardiac remodeling, atherosclerosis, restenosis, cardiac hypertrophy, cerebral ischemia, cerebral vasospasm, and erectile dysfunction.

[0079] In other embodiments, the ROCK2 inhibitors provided herein may be used in the treatment of a pulmonary disorder including idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, and asthma.

[0080] In other embodiments, the ROCK2 inhibitors provided herein may be used in the treatment of a central nervous system disorder, including neuronal degeneration or spinal cord injury, traumatic brain injury, cerebral cavernous malformation, Huntington’s disease, Parkinson’s disease, Alzheimer’s disease, Amyotrophic lateral sclerosis (ALS), or multiple sclerosis.

[0081] In other embodiments, the disclosure provides methods for the treatment of renal diseases including polycystic kidney disease, renal fibrosis and diabetic renal disease.21182500867.1

[0082] In other embodiments, the ROCK2 inhibitors provided herein may be used in the treatment of a metabolic disease including insulin resistance, hyperinsulinemia, type 2 diabetes, obesity, metabolic syndrome and glucose intolerance. The ROCK2 inhibitors may be used to effect weight loss and / or limit weight gain. In an embodiment, a ROCK2 inhibitor is used to reduce or prevent insulin resistance or restore insulin sensitivity.

[0083] In other embodiments, the ROCK2 inhibitors provided herein may be used in the treatment of an ocular disorder including ocular hypertension, age related macular degeneration (AMD; wet and dry), choroidal neovascularization (CNV), choroidal tumor, diabetic macular edema (DME), iris neovascularization, uveitis, glaucoma, primary openangle glaucoma, acute angle-closure glaucoma, pigmentary glaucoma, congenital glaucoma, normal tension glaucoma, secondary glaucoma, neo vascular glaucoma, geographic atrophy, and retinitis of prematurity (ROP).

[0084] In another embodiment, the disclosure provides methods for the treatment of sickle cell disease.

[0085] In other embodiments, the ROCK2 inhibitors provided herein may be used to treat (i.e., cure or reduce the severity of, etc.) viral infections, particularly coronavirus infections such as SARS-CoV-1, SARS-CoV-2, and MERS-CoV, and to treat or prevent the sequelae resulting from the viral infection, including the coronavirus infection such as SARS-CoV-1, SARS-CoV-2, and MERS-CoV. In some embodiments, the viral infection is a SARS-CoV-1 infection. In some embodiments, the viral infection is a SARS-CoV-2 infection. In some embodiments, the viral infection is a MERS-CoV infection. In embodiments, the sequelae include one or more of the group consisting of fatigue, dyspnea (difficulty breathing), cough, arthralgia (joint pain), myalgia, headache, chest pain, fever, palpitations, myocardial inflammation, ventricular dysfunction, stroke, pulmonary function abnormalities, fibrosis (such as pulmonary fibrosis), renal dysfunction rash, alopecia, olfactory and / or gustatory dysfunction, sleep dysregulation, cognitive impairment altered, memory impairment, depression, anxiety, changes in mood and combinations thereof. In embodiments, the sequelae include inflammation and / or fibrosis.Pharmaceutical Compositions

[0086] In one aspect, the present disclosure provides pharmaceutically acceptable compositions for use in the treatment of viral diseases which comprise a therapeutically-effective amount of one or more of the ROCK2 inhibitors provided in this disclosure, formulated together with one or more pharmaceutically acceptable carriers. As described 22182500867.1below, the pharmaceutical compositions of the present disclosure may be specially formulated for administration in solid or liquid form, including those adapted for the following: (1) oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; (2) parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; (3) topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin; (4) intravaginal or intrarectal administration, for example, as a suppository, pessary, cream or foam; (5) sublingual administration; (6) ocular administration; (7) transdermal administration; or (8) nasal administration.

[0087] The phrase “pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals with toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0088] The phrase “pharmaceutically-acceptable carrier” as used herein means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc magnesium, calcium or zinc stearate, or steric acid), solvent, or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ, or portion of the body, to another organ, or portion of the body. Each carrier should be compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) pH buffered solutions; (21) polyesters, polycarbonates and / or23182500867.1polyanhydrides; and (22) other non-toxic compatible substances employed in pharmaceutical formulations.

[0089] The compounds of this disclosure may be formulated with conventional carriers and excipients, which can be selected in accord with ordinary practice. Tablets can contain excipients, glidants, fillers, binders and the like. Aqueous formulations are prepared in sterile form, and when intended for delivery by other than oral administration generally can be isotonic. All formulations can optionally contain excipients such as those set forth in the “Handbook of Pharmaceutical Excipients” (1986). Excipients include ascorbic acid and other antioxidants, chelating agents such as EDTA, carbohydrates such as dextran, hydroxyalkylcellulose, hydroxyalkylmethylcellulose, stearic acid and the like.

[0090] While it is possible for the ROCK2 inhibitors disclosed herein (herein referred to as the “active ingredients”) to be administered alone, it may be preferable to present them as pharmaceutical formulations. The formulations, both for veterinary and for human use, of the disclosure comprise at least one active ingredient, as provided above, together with one or more acceptable carriers therefor and optionally other therapeutic ingredients, particularly those additional therapeutic ingredients as discussed herein.

[0091] The formulations include those suitable for the administration routes provided herein. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. Techniques and formulations generally are found in Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, Pa.). Such methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

[0092] Formulations of the present disclosure suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets, each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous or non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient may also be administered as a bolus, electuary or paste.

[0093] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally 24182500867.1mixed with a binder, lubricant, inert diluent, preservative, surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered active ingredient moistened with an inert liquid diluent. The tablets may optionally be coated or scored and optionally are formulated so as to provide slow or controlled release of the active ingredient therefrom.

[0094] For infections of the eye or other external tissues e.g. mouth and skin, the formulations are preferably applied as a topical solution, ointment or cream containing the active ingredient(s). The active ingredient may be present in an amount of, for example, 0.075 to 20% w / w (including active ingredient(s) in a range between 0.1% and 20% in increments of 0.1% w / w such as 0.6% w / w, 0.7% w / w, etc.), preferably 0.2 to 15% w / w and most preferably 0.5 to 10% w / w. When formulated in an ointment, the active ingredients may be employed with either a paraffinic or a water-miscible ointment base. Alternatively, the active ingredients may be formulated in a cream with an oil-in-water cream base.

[0095] If desired, the aqueous phase of the cream base may include, for example, at least 30% w / w of a polyhydric alcohol, i.e., an alcohol having two or more hydroxyl groups such as propylene glycol, butane 1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol (including PEG 400) and mixtures thereof. The topical formulations may desirably include a compound which enhances absorption or penetration of the active ingredient through the skin or other affected areas. Examples of such dermal penetration enhancers include dimethyl sulfoxide and related analogs.

[0096] The oily phase of the emulsions of this disclosure may be constituted from known ingredients in a known manner. While the phase may comprise merely an emulsifier (otherwise known as an emulgent), it desirably comprises a mixture of at least one emulsifier with a fat or an oil or with both a fat and an oil. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier which acts as a stabilizer. It is also preferred to include both an oil and a fat. Together, the emulsifier(s) with or without stabilizer(s) make up the so-called emulsifying wax, and the wax together with the oil and fat make up the so-called emulsifying ointment base which forms the oily dispersed phase of the cream formulations.

[0097] Emulsifying agents and emulsion stabilizers suitable for use in the formulation of the disclosure include Tween® 60, Span® 80, cetostearyl alcohol, benzyl alcohol, myristyl alcohol, glyceryl mono-stearate and sodium lauryl sulfate. Further emulsifying agents and emulsion stabilizers suitable for use in the formulation of the disclosure include Tween® 80.25182500867.1

[0098] The choice of suitable oils or fats for the formulation is based on achieving the desired properties. The cream should preferably be a non-greasy, non-staining and washable product with suitable consistency to avoid leakage from tubes or other containers. Straight or branched chain, mono- or dibasic alkyl esters such as di-isoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or a blend of branched chain esters known as Crodamol CAP may be used, the last three being preferred esters. These may be used alone or in combination depending on the properties required. Alternatively, high melting point lipids such as white soft paraffin and / or liquid paraffin or other mineral oils are used.

[0099] Pharmaceutical formulations according to the present disclosure comprise a combination according to the disclosure together with one or more pharmaceutically acceptable carriers or excipients and optionally other therapeutic agents. Pharmaceutical formulations containing the active ingredient may be in any form suitable for the intended method of administration. When used for oral use for example, tablets, troches, lozenges, aqueous or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs may be prepared. Compositions intended for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents including sweetening agents, flavoring agents, coloring agents and preserving agents, in order to provide a palatable preparation. Tablets containing the active ingredient in admixture with non-toxic pharmaceutically acceptable excipient which are suitable for manufacture of tablets are acceptable. These excipients may be, for example, inert diluents, such as calcium or sodium carbonate, lactose, calcium or sodium phosphate; granulating and disintegrating agents, such as maize starch, or alginic acid; binding agents, such as starch, gelatin or acacia; and lubricating agents, such as magnesium stearate, stearic acid or talc. Tablets may be uncoated or may be coated by known techniques including microencapsulation to delay disintegration and adsorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate alone or with a wax may be employed.

[0100] Formulations for oral use may be also presented as hard gelatin capsules where the active ingredient is mixed with an inert solid diluent, for example starch, mannitol, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin or olive oil.26182500867.1

[0101] Aqueous suspensions of the disclosure contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients include a suspending agent, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing or wetting agents such as a naturally-occurring phosphatide (e.g., lecithin), a condensation product of an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate), a condensation product of ethylene oxide with a long chain aliphatic alcohol (e.g., heptadecaethyleneoxycetanol), a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol anhydride (e.g., polyoxyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives such as ethyl or n-propyl p-hydroxy -benzoate, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, such as sucrose or saccharin. Further non-limiting examples of suspending agents include Cyclodextrin and Captisol (=Sulfobutyl ether betacyclodextrin; SEB-beta-CD).

[0102] Oil suspensions may be formulated by suspending the active ingredient in a vegetable oil, such as arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oral suspensions may contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents, such as those set forth above, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an antioxidant such as ascorbic acid.

[0103] Dispersible powders and granules of the disclosure suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those disclosed above. Additional excipients, for example sweetening, flavoring and coloring agents, may also be present.

[0104] The pharmaceutical compositions of the disclosure may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, a mineral oil, such as liquid paraffin, or a mixture of these. Suitable emulsifying agents include naturally-occurring gums, such as gum acacia and gum tragacanth, naturally-occurring phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweetening and flavoring agents. Syrups and elixirs may be formulated with 27182500867.1sweetening agents, such as glycerol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative, a flavoring or a coloring agent.

[0105] The pharmaceutical compositions of the disclosure may be in the form of a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butane-diol or prepared as a lyophilized powder. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils may conventionally be employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid may likewise be used in the preparation of injectables. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution isotonic sodium chloride solution, and hypertonic sodium chloride solution.

[0106] The amount of active ingredient that may be combined with the carrier material to produce a single dosage form can vary depending upon the host treated and the particular mode of administration. For example, a time-release formulation intended for oral administration to humans may contain approximately 1 to 1000 mg of active material compounded with an appropriate and convenient amount of carrier material which may vary from about 5 to about 95% of the total compositions (weight weight). The pharmaceutical composition can be prepared to provide easily measurable amounts for administration. For example, an aqueous solution intended for intravenous infusion may contain from about 3 to 500 pg of the active ingredient per milliliter of solution in order that infusion of a suitable volume at a rate of about 30 mL / hr can occur.

[0107] Formulations suitable for topical administration to the eye also include eye drops wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active ingredient. The active ingredient may be present in such formulations in a concentration of 0.5 to 20%, advantageously 0.5 to 10%, and particularly about 1.5% w / w.

[0108] Formulations suitable for topical administration in the mouth include lozenges comprising the active ingredient in a flavored basis, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert basis such as gelatin and glycerin, or28182500867.1sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

[0109] Formulations for rectal administration may be presented as a suppository with a suitable base comprising for example cocoa butter or a salicylate.

[0110] Formulations suitable for intrapulmonary or nasal administration have a particle size for example in the range of 0.1 to 500 microns, such as 0.5, 1, 30, 35 etc., which is administered by rapid inhalation through the nasal passage or by inhalation through the mouth so as to reach the alveolar sacs. Suitable formulations include aqueous or oily solutions of the active ingredient. Formulations suitable for aerosol or dry powder administration may be prepared according to conventional methods and may be delivered with other therapeutic agents such as compounds.

[0111] Formulations suitable for vaginal administration may be presented as suppositories, pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the active ingredient such carriers as are known in the art to be appropriate.

[0112] Formulations suitable for parenteral administration include aqueous and nonaqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents.

[0113] The formulations are presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injection, immediately prior to use. Extemporaneous injection solutions and suspensions are prepared from sterile powders, granules and tablets of the kind previously described. Preferred unit dosage formulations are those containing a daily dose or unit daily sub-dose, as herein above recited, or an appropriate fraction thereof, of the active ingredient.

[0114] The disclosure further provides veterinary compositions comprising at least one active ingredient as above defined together with a veterinary carrier therefor.

[0115] Veterinary carriers are materials useful for the purpose of administering the composition and may be solid, liquid or gaseous materials which are otherwise inert or acceptable in the veterinary art and are compatible with the active ingredient. These veterinary compositions may be administered orally, parenterally or by any other desired route.29182500867.1

[0116] Compounds of the disclosure are used to provide controlled release pharmaceutical formulations containing as active ingredient one or more compounds of the disclosure (“controlled release formulations”) in which the release of the active ingredient are controlled and regulated to allow less frequency dosing or to improve the pharmacokinetic or toxicity profile of a given active ingredient.

[0117] The patient receiving this treatment is any animal in need, including primates, in particular humans, and other mammals such as horse, cattle, swine and sheep; and poultry and pets in general.EXAMPLESExample 1

[0118] (6-(4-((4-( 1 H-Pyrazol-4-yl)phenyl)amino)-7, 8-dihy dro-5H-pyrano[4, 3 -d]pyrimidin-2-yl)-l -methyl- lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)m ethanone

[0119] Synthesis:Pd(PPh3)4, K2CO3step 1dioxane / H2Ostep 3182500867.1

[0120] Step 1: Synthesis of 4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl) aniline. To a solution of 4-bromoaniline (14.60 g, 84.87 mmol) and l-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (28.33 g, 101.85 mmol) in 1,4-dioxane / JLO (500 / 100 mL) was added K2CO3 (23.46 g, 169.75 mmol) and Pd(PPh3)4 (9.81 g, 8.49 mmol). The reaction mixture was stirred at 100 °C for 16 h under N2. The reaction mixture was cooled to room temperature and poured into water (800 mL), then extracted with EA (1000 mL x 3). The combined organic layers were washed with brine (800 mL x 3), dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by column chromatography (PE: EA=1:1) to afford 4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl) aniline (9 g, 43.6% yield) as a yellow solid. LC-MS (ESI): 244.3, [M+H]+. 'H NMR (400 MHz, DMSO-tfe) 88.05 (s, 1H), 7.72 (s, 1H), 7.25 (d, J = 8.4 Hz, 2H), 6.55 (d, J = 8.4 Hz, 2H), 5.35 (dd, J = 10.0, 2.1 Hz, 1H), 5.02 (s, 2H), 3.92 (d, J = 12.2 Hz, 1H), 3.62 (ddd, J = 21.0, 11.9, 7.0 Hz, 1H), 2.17 - 1.49 (m, 6H).

[0121] Step 2: Synthesis of 2-chloro-N-(4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)-7,8-dihydro-5H-pyrano[4,3-d]pyrimidin-4-amine. To a solution of 4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl) aniline (1.0 g, 4.11 mmol) and 2,4-dichloro-7,8-dihydro-5H-pyrano[4,3-d] pyrimidine (843 mg, 4.11 mmol) in / / -BuOH (30 mL) was added DIEA (1.59 g, 12.33 mmol). The reaction mixture was stirred at 100 °C for 16 h. The reaction mixture was cooled to room temperature and poured into water (50 mL), then extracted with EA (30 mL x 3). The combined organic layers were washed with brine (30 mL x 3), dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by column chromatography (PE: EA=20:ll) to afford 2-chloro-N-(4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)-7,8-dihydro-5H-pyrano[4,3-d] pyrimidin-4-amine (700 mg, 41.35% yield) as a light yellow solid. LC-MS (ESI): 412.3, [M+H]+. 'H NMR (400 MHz, DMSO-t / 6) 5 8.71 (s, 1H), 8.31 (s, 1H), 7.93 (s, 1H), 7.61 (d, J = 8.7 Hz, 2H), 7.56 (d, J = 8.7 Hz, 2H), 5.41 (dd, J = 9.9, 2.2 Hz, 1H), 4.61 (s, 2H), 3.97 - 3.88 (m, 3H), 3.70 - 3.59 (m, 1H), 2.71 (t, J = 5.5 Hz, 2H), 2.18 - 1.48 (m, 6H).

[0122] Step 3: Synthesis of (3,3-difluoroazetidin-l-yl)(l-methyl-6-(4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydro-5H-pyrano[4,3-d]pyrimidin-2-yl)-lH-indol-2-yl)methanone. To a solution of 2-chloro-N-(4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl) phenyl)-7,8-dihydro-5H-pyrano[4,3-d]pyrimidin-4-amine (400 mg, 0.97 mmol) and (3,3-difluoroazetidin-l-yl) (l-methyl-6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-indol-2-yl) methanone (438 mg, 1.17 mmol) in dioxane / ^O (10 / 2 mL) was added K2CO3 (268 mg, 1.94 mmol) and Pd(PPh3)4(l 12 mg, 0.10 mmol). The reaction mixture was 31182500867.1stirred at 100 °C for 16 h under N2. The reaction mixture was cooled to room temperature and poured into water (20 mL), then extracted with EA (20 mLz3 ). The combined organic layers were washed with brine (20 mL x 3), dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by column chromatography (PE: EA=5:3) to afford (3,3-difluoroazetidin-l-yl)(l-methyl-6-(4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl) amino)-7,8-dihydro-5H-pyrano[4,3-d]pyrimidin-2-yl)-lH-indol-2-yl) methanone (310 mg, 51.02% yield) as a light yellow solid. LC-MS (ESI): 626.5, [M+H]+.XHNMR (400 MHz, DMSO-de) 88.48 (s, 1H), 8.40 (s, 1H), 8.33 (s, 1H), 8.13 (d, J = 8.4 Hz, 1H), 7.95 (s, 1H), 7.82 (d, J = 8.6 Hz, 2H), 7.68 (t, J = 9.2 Hz, 3H), 7.05 (s, 1H), 5.42 (dd, J = 9.9, 2.1 Hz, 1H), 4.67 (d, J = 46.5 Hz, 6H), 4.10 - 3.97 (m, 6H), 3.72 - 3.57 (m, 1H), 2.87 (d, J = 5.3 Hz, 2H), 2.13 (dd, J= 18.6, 6.7 Hz, 1H), 2.02 - 1.91 (m, 2H), 1.70 (d, J = 11.8 Hz, 1H), 1.57 (d, J = 3.6 Hz, 2H).

[0123] Step 4: Synthesis of (6-(4-((4-(lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydro-5H-pyrano[4,3-d]pyrimidin-2-yl)-l -methyl- IH-indol -2 -yl)(3,3-difluoroazeti din- l-yl)methanone. To a solution of (3,3-difluoroazetidin-l-yl) (l-methyl-6-(4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl) phenyl) amino)-7,8-dihydro-5H-pyrano[4,3-d] pyrimidin-2-yl)-lH-indol-2-yl) methanone (310 mg, 0.50 mmol) in MeOH (2 mL) was added 4 M HC1 in EA (1 mL). The reaction mixture was stirred at 25 °C for 16 h. The reaction mixture was evaporated to dryness and the residue was portioned between EA and saturated solution of NaHCOs and extracted twice with EA. The combined organic phases were dried over sodium sulfate, filtered, and evaporated to dryness. The crude material was triturated with ACN, then repurified by prep-HPLC (Cl 8 prep-column) to provide (6-(4-((4-(lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydro-5H-pyrano[4,3-d]pyrimidin-2-yl)-l -methyl- lH-indol-2-yl)(3,3-difluoroazetidin-l-yl) methanone (140 mg, 52.18% yield) as a white solid. LC-MS (ESI): 542.4, [M+H]+. 'H NMR (400 MHz, DMSO-d6) 6 12.91 (s, 1H), 8.47 (s, 1H), 8.40 (s, 1H), 8.18 - 8.12 (m, 2H), 7.94 (s, 1H), 7.80 (d, J = 8.4 Hz, 2H), 7.70 - 7.64 (m, 3H), 7.05 (s, 1H), 4.86 - 4.60 (m, 6H), 4.00 (s, 5H), 2.86 (s, 2H).Example 232182500867.1

[0124] (6-(4-((4-(lH-Pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l -methyl- lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)m ethanone

[0125] Synthesis:THP

[0126] Step 1: Synthesis of tert-butyl 2-chloro-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate. To a solution of tert-butyl 2,4-dichloro-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate (2.5 g, 8.22 mmol) in BuOH (30 mL) was added 4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)aniline (2.0 g, 8.22 mmol) and DIPEA (4.2 mL, 24.7 mmol). The resulting mixture was stirred for 16 hours at 110 °C. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography (PE: EA=3:1) to afford tert-butyl 2-chloro-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate (3.2 g, 76.19% yield) as a yellow solid. LC-MS (ESI): 511.2, [M+H]+.

[0127] Step 2: Synthesis of tert-butyl 2-(2-(methoxycarbonyl)-l-methyl-lH-indol-6-yl)-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3- 33182500867.1d]pyrimidine-6(5H)-carboxylate. To a stirred solution of tert-butyl 2-chloro-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate (1 g, 1.96 mmol) in H2O (2 mL) and 1,4-dioxane (10 mL) was added methyl l-methyl-6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-indole-2-carboxylate (800 mg, 2.5 mmol), K2CO3 (350 mg, 2.5 mmol) and Pd(dppf)C12 (250 mg, 0.38 mmol) at room temperature. The resulting mixture was stirred for 16 hours at 100 °C. The desired product could be detected by LCMS. The reaction mixture was cooled to room temperature and poured into water (10 mL), then extracted with EA (20 mL *3). The combined organic layers were washed with brine (20 mL *3), dried over Na₂SO₄ and concentrated under reduced pressure. The residue was purified through column chromatography (SiCh, PE / EA=3 / 2) to afford tert-butyl 2-(2-(methoxycarbonyl)-l-methyl-lH-indol-6-yl)-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido [4, 3-d] pyrimidine-6(5H) -carboxylate (1 g, 83.33% yield) as a yellow oil. LCMS (ESI): 664.4, [M+l]+.

[0128] Step 3: Synthesis of 6-(6-(tert-butoxycarbonyl)-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lH-indole-2-carboxylic acid. To a solution of tert-butyl 2-(2-(methoxycarbonyl)-l-methyl-lH-indol-6-yl)-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate (800 mg, 1.2 mmol) in MeOH (2 mL), THF (2 mL) and H2O (2 mL) was added lithium hydroxide monohydrate (151 mg, 3.6 mmol) at 25 °C. The resulting mixture was stirred for Ih. The desired product could be detected by LCMS. The reaction mixture was cooled to room temperature and poured into water (10 mL), adjusted pH to 3-4, then extracted with EA (20 mL x 3). The combined organic layers were washed with brine (20 mL x 3), dried over Na₂SO₄ and concentrated under reduced pressure. The residue was purified through column chromatography (SiO2, PE / EA=7 / 1) to afford 6-(6-(tert-butoxycarbonyl)-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lH-indole-2-carboxylic acid (500 mg, 63.80% yield) as a yellow solid. LC-MS (ESI): 650.4, [M+l]+.

[0129] Step 4: Synthesis of tert-butyl 2-(2-(3,3-difluoroazetidine-l-carbonyl)-l-methyl-lH-indol-6-yl)-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate. To a solution of 6-(6-(tert-butoxycarbonyl)-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)- 5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lH-indole-2-carboxylic acid (300 mg, 0.46 mmol) in DMF (3 mL) was added 3, 3 -difluoroazeti dine, hydrochloride (60 mg, 0.4634182500867.1mmol) and DIEA (179 mg, 1.38 mmol) and HATU (352 mg, 0.92 mmol) at RT. The resulting mixture was stirred for 1 hour at 25 °C. Desired product could be detected by LCMS. The reaction mixture was cooled to room temperature, poured into water (10 mL), and then extracted with EA (20 mL x 3). The combined organic layers were washed with brine (20 mL x 3), dried over Na₂SO₄ and concentrated under reduced pressure. The residue was purified through column chromatography (SiCh, PEZEA=3 / 7) to afford tert-butyl 2-(2-(3,3-difluoroazetidine-l-carbonyl)-l-methyl-lH-indol-6-yl)-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate (250 mg, 74.85% yield) as a yellow solid. LC-MS (ESI): 725.6, [M+l]+.

[0130] Step 5: Synthesis of tert-butyl 2-(2-(3,3-difluoroazetidine-l-carbonyl)-l-methyl-lH-indol-6-yl)-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate. A solution of tert-butyl 2-(2-(3,3-difluoroazetidine-l-carbonyl)-l-methyl-lH-indol-6-yl)-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate (300 mg, 0.41 mmol) in hydrochloric acid (2 mL) was stirred for 1 hour at 25 °C. The desired product could be detected by LCMS. The resulting mixture was concentrated under vacuum to afford a crude product, which was directly used without further purification. LC-MS (ESI): 541.3, [M+l]+.Example 3

[0131] (6-(4-((4-(lH-Pyrazol-4-yl)phenyl)amino)-6-methyl-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l -methyl- lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)m ethanone

[0132] Synthesis:35182500867.1

[0133] Step 1: Synthesis of (6-(4-((4-(lH-pyrazol-4-yl)phenyl)amino)-6-methyl-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone (Compound 3). To a solution of (6-(4-((4-(lH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone (40 mg, 0.06 mmol) in MeOH (20 mL) and AcOH (0.2 mL) was added formaldehyde (2 mL, 0.6 mmol) and sodium triacetoxyborohydride (30 mg, 0.14 mmol) at room temperature. The resulting mixture was stirred for 3 hours at 25 °C. Desired product could be detected by LCMS. The crude product was purified by prep-HPLC with the following conditions: Sunfire C18 19*250*10 um; Mobile Phase A: 0.05%TFA / H20, B1ACN; flow rate: 20ml / min; gradient: 26%-27%; Retention Time: 7.5-9.0 min of 16 min to afford (6-(4-((4-(lH-pyrazol-4-yl)phenyl)amino)-6-methyl-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l -methyl- lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)m ethanone (26 mg, 65% yield) as a yellow solid. LC-MS (ESI): 555.3, [M+H]+. 'H NMR (400 MHz, DMSO-tL) 5 8.45 (d, J = 14.5 Hz, 2H), 8.22 (s, 1H), 8.14 - 8.11 (m, 1H), 8.06 (s, 1H), 7.83 (d, J = 8.6 Hz, 2H), 7.70 - 7.63 (m, 3H), 7.04 (s, 1H), 4.58 (s, 4H), 4.01 (s, 3H), 3.51 (s, 2H), 2.87 (d, J = 5.2 Hz, 2H), 2.75 (d, J = 5.4 Hz, 2H), 2.48 (s, 3H).Example 4

[0134] (6-(4-((4-(lH-Pyrazol-4-yl)phenyl)amino)-6-cyclopropyl-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone182500867.1

[0135] Synthesis:

[0136] Step 1: Synthesis of (6-(4-((4-(lH-pyrazol-4-yl)phenyl)amino)-6-cyclopropyl-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone (Compound 4). To a solution of tert-butyl 2-(2-(3,3-difluoroazetidine-l-carbonyl)-l-methyl-lH-indol-6-yl)-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate(200 mg, 0.37 mmol) in MeOH (2 mL) and AcOH (0.5 mL) was added (1 -ethoxy cy cl opropoxy)-trimethylsilane (193 mg, 1.1 mmol) for 0.5 hour at 25 °C. To the resulting mixture was added NaBHsCN (235 mg, 1.1 mmol) and the mixture was stirred for 1 hour at 25 °C.Desired product could be detected by LCMS. The crude product was purified by prep-HPLC with the following conditions: Sunfire C18 19*250*10 um; Mobile Phase A:0.05%TFA / H20, BlACN;flow rate: 20 ml / min; gradient: 27%-27%; Retention Time: 8.1-9.7 min of 16 min to afford (6-(4-((4-(lH-pyrazol-4-yl)phenyl)amino)-6-cyclopropyl-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone (16.49 mg, 7.70% yield) as a yellow solid. LC-MS (ESI): 581.4, [M+H]+.TH NMR (400 MHz, CD3OD) 58.42 (s, 1H), 8.03 (s, 2H), 7.97 (d, J = 8.4 Hz, 1H), 7.79 (d, J = 8.5 Hz, 1H), 7.72 (s, 4H), 7.02 (s, 1H), 4.62 (d, J = 38.8 Hz, 4H), 4.16 (s, 2H), 4.04 (s, 3H), 3.48 - 3.45 (m, 2H), 3.18 (d, J = 5.9 Hz, 2H), 2.54 (s, 1H), 0.87 (d, J = 13.1 Hz, 4H).Example 5182500867.1

[0137] (6-(4-((4-(lH-Pyrazol-4-yl)phenyl)amino)-6-(2-methoxyethyl)-5,6,7,8- tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lH-indol-2-yl)(3,3-difluoroazetidin-l- yl)methanone

[0138] Synthesis:

[0139] Step 1: tert-Butyl 2-chloro-4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-lH-pyrazol-4- yl)phenyl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5J7)-carboxylate. To a solution of tert-butyl 2,4-dichloro-7,8-dihydropyrido[4,3-d]pyrimidine-6(5rt)-carboxylate (15 g, 49.315 mmol) and 4-(l-(tetrahydro-2J / -pyran-2-yl)-lJH-pyrazol-4-yl)aniline (10.91 g, 44.831 mmol) in 2-methylpropan-2-ol (180 mL) was added DIEA (17.4 g, 134.49 mmol). The mixture was stirred at 110 °C for 16 hrs. The reaction mixture was concentrated, diluted with ice-water and extracted with EA (3^50 mL) and the combined organic layers were dried over Na₂SO₄ and concentrated, then purified by column chromatography on silica gel eluting with methanol in di chloromethane (from 0 % to 10 %) to afford tert-butyl 2-chloro-4-((4-(l- (tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3- d]pyrimidine-6(5H)-carboxylate (16 g, 31.31 mmol, 69.8 % yield) as a yellow solid. LC-MS (ESI): 511.1, [M+H]+.38182500867.1

[0140] Step 2: tert-Butyl 2-(2-(methoxycarbonyl)-l-methyl-lH-indol-6-yl)-4-((4-(l-(tetrahydro-2 / / -pyran-2-yl)-l / / -pyrazol-4-yl)phenyl)arnino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5J7)-carboxylate. Methyl l-methyl-6-(4, 4,5, 5-tetramethyl- 1,3,2-dioxaborolan-2-yl)-lH-indole-2-carboxylate (2.22 g, 7.05 mmol), tert-butyl 2-chloro-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5J7)-carboxylate (3.0 g, 5.871 mmol) and anhydrous potassium carbonate (1.62 g, 11.74 mmol) were dissolved in 1,4-dioxane (60 mL) and H2O (12 mL). The reaction mixture was purged with argon and then l,l'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride dichloromethane complex (0.19 g, 0.29 mmol) was added. The reaction mixture was stirred at 100 °C for overnight. TLC analysis showed full conversion of the starting material. The reaction mixture was dissolved in EtOAc, filtered, concentrated and purified using column chromatography (0-20% EtOAc in DCM, gradient elution) to give tert-butyl 2-(2-(methoxycarbonyl)-l-methyl-lH-indol-6-yl)-4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate (2.8 g, 4.22 mmol, 71.8% yield). LC-MS (ESI): 664.2, [M+H]+.

[0141] Step 3: Methyl l-methyl-6-(4-((4-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-lJ / -indole-2-carboxylate. To a solution of tert-butyl 2-(2-(methoxycarbonyl)-l-methyl-lJ / -indol-6-yl)-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (2.0 g, 3.12 mmol) in Methanol (2 mL) and dichloromethane (10 mL) was added ZnBn (10.16 g, 45.18 mmol). The reaction was stirred at 50 °C for 3 h. LCMS showed the starting material disappeared, and the major product was the desired. The reaction was filtered and the filtrate was concentrated, then purified by column chromatography on silica gel eluting with methanol in di chloromethane (from 0 % to 10 %) to give methyl l-methyl-6-(4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-lJ / -indole-2-carboxylate (650 mg, 1.06 mmol, 34.1% yield) as a yellow solid. LC-MS (ESI): 564.2, [M+H]+.

[0142] Step 4: Methyl 6-(6-(2-m ethoxy ethyl)-4-((4-(l-(tetrahydro-2 / / -pyran-2-yl)-l / / -pyrazol-4-yl)phenyl)amino)-5, 6,7, 8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l -methyl- \H-indole-2-carboxylate. To a solution of methyl l-methyl-6-(4-((4-( l-(tetrahydro-2 / / -pyran-2-yl)-17 / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-lJH-indole-2-carboxylate (600 mg, 1.064 mmol) and l-bromo-2-m ethoxy ethane (296 mg, 2.128 mmol) in N,N-dimethylformamide (10 mL) was added K2CO3 (440 mg, 3.191 mmol). The mixture was stirred at 85 °C for 6 hrs. The reaction mixture was diluted with ice-water and 39182500867.1extracted with EA (3^50 mL) and the combined organic layers were dried overlSfeSCU and concentrated, then purified by column chromatography on silica gel eluting with methanol in di chloromethane (from 0% to 10 %) to afford methyl 6-(6-(2-methoxyethyl)-4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lZ / -indole-2-carboxylate (260 mg, 0.419 mmol, 39.4 % yield) as a yellow solid. LC-MS (ESI): 622.1, [M+H]+.

[0143] Step 5: 6-(6-(2-Methoxyethyl)-4-((4-(l-(tetrahydro-2Z / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5, 6,7, 8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l -methyl- lZ / -indole-2-carboxylic acid. To a solution of methyl 6-(6-(2-methoxyethyl)-4-((4-(l-(tetrahydro-2Z7-pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-1 -methyl- l / / -indole-2-carboxylate (260 mg, 0.419 mmol) in methanol (5 mL) and THF (5 mL) and H2O (1.5 mL) were added LiOH (50 mg, 2.093 mmol). The mixture was stirred at room temperature for 12 hrs. The reaction was adjusted pH to 3~4, and extracted with EA (3x50 mL) and the combined organic layers were dried over Na₂SO₄ and concentrated to afford 6-(6-(2-methoxyethyl)-4-((4-( l-(tetrahydro-27 / -pyran-2-yl)- IT / -pyrazol-4-yl)phenyl)amino)-5, 6,7, 8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l -methyl- l / Z-indole-2-carboxylic acid (190 mg, 0.31 mmol, 74.6 % yield) as a yellow solid. LC-MS (ESI): 608.2, [M+H]+.

[0144] Step 6: (3,3-Difluoroazetidin-l-yl)(6-(6-(2-methoxyethyl)-4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-1 -methyl- IT / -indol -2 -yl)methanone. To a solution of 6-(6-(2-methoxyethyl)-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-IT / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lZ / -indole-2-carboxylic acid (190 mg, 0.313 mmol) in DMF (2 mL) were added DIEA (121 mg, 0.937 mmol), HATU (238 mg, 0.625 mmol) and 3,3-difluoroazetidine (61 mg, 0.469 mmol). The mixture was stirred at 25 °C for 2 hours. The reaction mass was diluted with ice-water and extracted with EtOAc (3x8 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated and was purified by column chromatography on silica gel eluting with methanol in dichloromethane (from 0% to 10 %) to give (3,3-difluoroazetidin-l-yl)(6-(6-(2-methoxyethyl)-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-IT / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l -methyl- 177-indol-2-yl)methanone (170.00 mg, 0.249 mmol, 79.5 % yield) as a yellow solid. LC-MS (ESI): 683.2, [M+H]+.

[0145] Step 7: (6-(4-((4-(lZ / -Pyrazol-4-yl)phenyl)amino)-6-(2-methoxyethyl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-U7-indol-2-yl)(3,3-difluoroazeti din-1- 40182500867.1yl)methanone. To a solution of (3,3-difluoroazetidin-l-yl)(6-(6-(2-methoxyethyl)-4-((4-(l-(tetrahydro-2 / / -pyran-2-yl)-l / / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l -methyl- U / -indol-2-yl)methanone (170.00 mg, 0.249 mmol) in MeOH (5 mL) was added HC1 (2 mL). The mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated and adjusted pH to 9 by aq. NaHCOs and extracted with EtOAc (3x8 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (Waters 3767QDA Column: XBridge C18, 19x250mm, 10 pm; Mobile Phase A: 0.1% NH4OH / H2O, B: ACN; flow rate: 20 mL / min; gradient: 46-46%; Retention Time: 6.2-7. Imin of 17min) to afford (6-(4-((4-(U / -pyrazol-4-yl)phenyl)amino)-6-(2-methoxyethyl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-U7-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone (65.59 mg, 0.109 mmol, 43.7 % yield) as a white solid. LC-MS (ESI): 599.5, [M+H]+.XHNMR (400 MHz, DMSO-d6) 58.46 (s, 1H), 8.42 (s, 1H), 8.32 (s, 1H), 8.10 (dd, J= 15.1, 13.9 Hz, 3H), 7.81 (d, J= 8.6 Hz, 2H), 7.66 (t, J= 9.4 Hz, 3H), 7.04 (s, 1H), 4.60 (s, 4H), 4.01 (s, 3H), 3.63 - 3.59 (m, 4H), 3.31 (s, 3H), 2.85 (s, 4H), 2.79 (t, J= 6.0 Hz, 2H).Example 6

[0146] (6-(4-((4-(lH-Pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l -methyl- lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)m ethanone

[0147] Synthesis:41182500867.1dioxane, 100°C, 6 h01 03

[0148] Step 1: tert-Butyl 2-chloro-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-IT / -pyrazol-4-yl)phenyl)amino)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate. To a mixture of tert-butyl 2,4-dichloro-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (27.5 g, 90.42 mmol) and 4-(l-(tetrahydro-2J / -pyran-2-yl)-lJH-pyrazol-4-yl)aniline (20.0 g, 82.20 mmol) in w-BuOH (220 mL) were added DIEA (31.9 g, 246.61 mmol), The reaction mixture was stirred at 100 °C for 16 hours under Ar atmosphere. The reaction mixture was cooled to room temperature and quenched by saturated ammonia chloride solution (300 mL). Then the mixture was extracted with EtOAc (500 mL x 3) and the combined organic layers were washed by water (300 mL) and brine (300 mL). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue, which was purified by column chromatography on silica gel eluting with ethyl acetate (from 0 % to 50 %) in petroleum ether to give tert-butyl 2-chloro-4-((4-( l-(tetrahydro-27 / -pyran-2-yl)- IT / -pyrazol-4-yl)phenyl)amino)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (30.0 g, 58.71 mmol, 64.9 % yield) as a yellow solid. LC-MS (ESI):510.9, [M+H]+.

[0149] Step 2: tert-Butyl 2-(2-(3,3 -difluoroazetidine- 1 -carbonyl)- 1 -methyl- U / -indol-6-yl)-4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate. To a mixture of tert-butyl 2-chloro-4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (3.0 g, 5.87 mmol) and (3,3-difluoroazetidin-1-yl)(1-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)methanone (2.6 g, 7.05 mmol)42182500867.1in dioxane (60 mL) and water (12 mL) were added K2CO3 (1.6 g, 11.74 mmol) and Pd(dtpbf)C12 (190 mg, 0.29 mmol). The mixture was stirred at 100 °C for 6 hours under Ar atmosphere. The reaction mass was diluted with DCM (300 mL) and washed by water (100 mL) and brine (100 mL). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue, which was purified by column chromatography on silica gel eluting with methanol (from 0 % to 8 %) in dichloromethane to give tert-butyl 2-(2-(3,3-difluoroazetidine-l-carbonyl)-l-methyl-U / -indol-6-yl)-4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-U / -pyrazol-4-yl)phenyl)amino)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (2.7 g, 3.73 mmol, 63.4 % yield) as a yellow solid. LC-MS (ESI): 725.2, [M+H]+.

[0150] Step 3: (6-(4-((4-(U / -Pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-U / -indol-2-yl)(3,3-difluoroazetidin-l-yl)m ethanone. To a solution of tert-butyl 2-(2-(3, 3 -difluoroazeti dine- 1 -carbonyl)- 1 -methyl- U / -indol-6-yl)-4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-U / -pyrazol-4-yl)phenyl)amino)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (2.0 g, 2.76 mmol) in MeOH (20 mL) was added HC1 (g) in dioxane (3.4 mL, 13.79 mmol). The mixture was stirred at 25 °C for 2 hours. The reaction was concentrated and diluted with DCM (400 mL). Then the mixture was adjusted pH to 7-8 by saturated NaHCOs and washed with brine (100 mL). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue which was purified by prep-HPLC (Waters 3767QDA Column: XBridge C18, 19*250mm, 10 pm; Mobile Phase A: 0.1% NH4OH / H2O, B: ACN; flow rate: 20 mL / min; gradient: 35-35%; Retention Time: 6.7-8.0min of 17min) to afford (6-(4-((4-(17 / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-U / -indol-2-yl)(3,3-difluoroazetidin- l-yl)methanone (1.0 g, 1.85 mmol, 67.0 % yield) as a white solid. LC-MS (ESI): 541.3, [M+H]+.XHNMR (400 MHz, DMSO-d6) 5 12.90 (s, 1H), 8.45 (d, J= 4.7 Hz, 2H), 8.13 (dd, J= 8.4, 1.1 Hz, 2H), 7.95 (s, 1H), 7.86 (d, J= 8.6 Hz, 2H), 7.66 (dd, J= 14.3, 8.5 Hz, 3H), 7.04 (s, 1H), 5.14 -4.26 (m, 4H), 4.01 (s, 3H), 3.81 (s, 2H), 3.06 (t, J= 5.4 Hz, 2H), 2.60 (s, 2H).Example 743182500867.1

[0151] (6-(4-((4-(lH-Pyrazol-4-yl)phenyl)amino)-7-methyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l -methyl- lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)m ethanone

[0152] Synthesis:

[0153] Step 1: (6-(4-((4-(U / -Pyrazol-4-yl)phenyl)amino)-7-methyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-177-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone. To a mixture of (6-(4-((4-(U / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-177-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone (240 mg, 0.44 mmol) and 37 % formaldehyde (180 mg, 2.22 mmol) in MeOH (15 mL) were added HO Ac (0.2 mL). The mixture was stirred at room temperature for 1 h, then NaBHsCN (84 mg, 1.33 mmol) was added. The reaction mixture was stirred at room temperature for 16 h, diluted with water (20 mL) and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue, which was purified by column chromatography on silica gel eluting with methanol (from 0% to 10 %) in dichloromethane and Prep-HPLC (Waters 3767QDA Column: XBridge C18, 19*250mm, 10 pm; Mobile Phase A: 0.1% NH4HCO3 / H2O, B: ACN; flow rate: 20 mL / min; gradient: 46-46%; Retention Time: 6.9-8.2min of 17min) to give (6-(4-((4-(U / -pyrazol-4-yl)phenyl)amino)-7-methyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-UT-indol -2 -yl)(3, 3 -difluoroazeti din- l-yl)m ethanone (86 mg, 0.16 mmol, 35.2 % yield) as a yellow solid. LC-MS (ESI): 555., [M+H]+.XHNMR (400 MHz, DMSO-d6) 5 12.90 (s, 1H), 8.51 (s, 1H), 8.46 (s, 1H), 8.18 (s, 1H), 8.12 (d, J= 8.4 Hz, 1H), 7.93 (s, 1H), 7.86 (d, J= 8.644182500867.1Hz, 2H), 7.67 (dd, J= 14.4, 8.5 Hz, 3H), 7.05 (s, 1H), 4.61 (s, 4H), 4.01 (s, 3H), 3.51 (s, 2H), 2.70 (d, J= 22.9 Hz, 4H), 2.41 (s, 3H).Example 8

[0154] (6-(4-((4-(lH-Pyrazol-4-yl)phenyl)amino)-7-(2-fluoroethyl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone

[0155] Synthesis:

[0156] Step 1: tert-Butyl 2-chloro-4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-U / -pyrazol-4-yl)phenyl)amino)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate. To a mixture of tert-butyl 2,4-dichloro-5,8-dihydropyrido[3,4-d]pyrimidine-7(6rt)-carboxylate (27.5 g, 90.4245182500867.1mmol) and 4-(l-(tetrahydro-2J / -pyran-2-yl)-lJH-pyrazol-4-yl)aniline (20.0 g, 82.20 mmol) in / / -BuOH (220 mL) were added DIEA (31.9 g, 246.61 mmol). The reaction mixture was stirred at 100 °C for 16 hours under Ar atmosphere. The reaction mixture was cooled to room temperature and quenched by saturated ammonia chloride solution (300 mL). Then the mixture was extracted with EtOAc (500 mL x 3) and the combined organic layers were washed by water (300 mL) and brine (300 mL). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue, which was purified by column chromatography on silica gel eluting with ethyl acetate (from 0% to 50 %) in petroleum ether to give tert-butyl 2-chloro-4-((4-( l-(tetrahydro-2rt-pyran-2-yl)- lrt-pyrazol-4-yl)phenyl)amino)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (30.0 g, 58.71 mmol, 64.9 % yield) as a yellow solid. LC-MS (ESI):510.9, [M+H]+.

[0157] Step 2: tert-Butyl 2-(2-(methoxycarbonyl)-l-methyl-17 / -indol-6-yl)-4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate. Methyl l-methyl-6-(4, 4,5, 5-tetramethyl- 1,3,2-dioxaborolan-2-yl)-17 / -indole-2-carboxylate (2.22 g, 7.05 mmol), tert-butyl 2-chloro-4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (3.0 g, 5.871 mmol) and potassium carbonate anhydrous (1.62 g, 11.74 mmol) were dissolved in dioxane (60 mL) and H2O (12 mL). The reaction mixture was purged with argon and then l,l'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride dichloromethane complex (0.19 g, 0.29 mmol) was added. The reaction mixture was stirred at 100 °C overnight. TLC analysis showed full conversion of the starting material. The reaction mixture was dissolved in EtOAc, filtered, concentrated and purified using column chromatography (0-20% EtOAc in DCM, gradient elution) to give tertbutyl 2-(2-(methoxycarbonyl)-l-methyl-lJH-indol-6-yl)-4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-17 / -pyrazol-4-yl)phenyl)amino)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate (2.8 g, 5.72 mmol, 97.5% yield). LC-MS (ESI): 664.2, [M+H]+.

[0158] Step 3: Methyl l-methyl-6-(4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-lJH-indole-2-carboxylate. To a solution of tert-butyl 2-(2-(methoxycarbonyl)-l-methyl-17 / -indol-6-yl)-4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (680 mg, 1.02 mmol) in methanol (10 mL) and dichloromethane (10 mL) was added ZnBn (2.31 g, 10.2 mmol). The reaction was stirred at 50 °C for 3 h. LCMS showed the starting material disappeared and the major product was the desired product. The reaction was filtered and the filtrate was concentrated, then purified by 46182500867.1column chromatography on silica gel eluting with methanol (from 0% to 10 %) in di chloromethane to give methyl l-methyl-6-(4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-lJ / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-U / -indole-2-carboxylate (420 mg, 0.74 mmol, 73.0% yield) as a yellow solid. LC-MS (ESI): 564.2, [M+H]+.

[0159] Step 4: Methyl 6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l -methyl- 1H-indole-2-carboxylate. To a solution of methyl l-methyl-6-(4-((4-( l-(tetrahydro-27 / -pyran-2-yl)-17 / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-lJH-indole-2-carboxylate (500 mg, 0.887 mmol) and l-bromo-2-fluoroethane (225 mg, 1.77 mmol) in N,N-dimethylformamide (9 mL) was added K2CO3 (367 mg, 2.66 mmol). The mixture was stirred at 80 °C for 2 hrs. The reaction mixture was diluted with ice-water and extracted with EA (3x50 mL) and the combined organic layers were dried over Na₂SO₄, concentrated, and purified by column chromatography on silica gel eluting with methanol (from 0% to 10 %) in dichloromethane to afford methyl 6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-lJ / -indole-2-carboxylate (310 mg, 0.508 mmol, 57.3 % yield) as a yellow solid. LC-MS (ESI): 610.2, [M+H]+.

[0160] Step 5: 6-(7-(2-Fluoroethyl)-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5, 6,7, 8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l -methyl- UT-indole-2-carboxylic acid. To a solution of methyl 6-(7-(2-fluoroethyl)-4-((4-( l-(tetrahydro-27 / -pyran-2-yl)-lJ / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-lJ / -indole-2-carboxylate (340 mg, 0.558 mmol) in methanol (5 mL) and THF (5 mL) and H2O (1.5 mL) were added LiOH. EEO (117 mg, 2.79 mmol). The mixture was stirred at room temperature for 12 hrs. The reaction was adjusted pH to 3~4, extracted with EA (3x50 mL) and the combined organic layers were dried over Na₂SO₄ and concentrated to afford 6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-17 / -indole-2-carboxylic acid (280 mg, 0.47 mmol, 84.2 % yield) as a yellow solid. LC-MS (ESI): 596.3, [M+H]+.

[0161] Step 6: (3,3-Difluoroazetidin-l-yl)(6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-lJ / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-1 -methyl- UT-indol -2 -yl)methanone. To a solution of 6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-lJ / -indole-2-carboxylic acid (150 mg, 0.252 mmol) in DMF (247182500867.1mL) were added DIEA (97.5 mg, 0.756 mmol), HATU (192 mg, 0.504 mmol) and 3,3-difluoroazetidine (48.9 mg, 0.378 mmol). The mixture was stirred at 25 °C for 2 hours. The reaction mixture was diluted with ice-water and extracted with EtOAc (3x8 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography on silica gel eluting with methanol (from 0% to 10 %) in dichloromethane to give (3,3-difluoroazetidin-l-yl)(6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-U / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l -methyl- U / -indol-2-yl)methanone (105.00 mg, 0.156 mmol, 61.9 % yield) as a yellow solid. LC-MS (ESI): 671.2, [M+H]+.

[0162] Step 7: (6-(4-((4-(U / -Pyrazol-4-yl)phenyl)amino)-7-(2-fluoroethyl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-177-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone. To a solution of (3,3-difluoroazetidin-l-yl)(6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-U / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l -methyl- U / -indol-2-yl)methanone (105.00 mg, 0.156 mmol) in MeOH (5 mL) was added HC1 (2 mL). The mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated and adjusted pH to 9 by aq. NaHCOs and extracted with EtOAc (3x8 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (Waters 3767QDA Column: XBridge C18, 19*250mm, 10 pm; Mobile Phase A: 0.1% NH4OH / H2O, B: ACN; flow rate: 20 mL / min; gradient: 46-46%; Retention Time: 6.5-8.0min of 17min) to afford (6-(4-((4-(U / -pyrazol-4-yl)phenyl)amino)-7-(2-fluoroethyl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-U7-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone (38.47 mg, 0.0656 mmol, 42.0 % yield) as a white solid. LC-MS (ESI): 587.4, [M+H]+.XHNMR (400 MHz, DMSO-d6) 5 12.90 (s, 1H), 8.52 (s, 1H), 8.46 (s, 1H), 8.12 (d, J= 9.6 Hz, 2H), 8.01 - 7.91 (m, 1H), 7.86 (d, J= 8.6 Hz, 2H), 7.67 (dd, J= 13.5, 8.6 Hz, 3H), 7.05 (s, 1H), 4.98 - 4.42 (m, 6H), 4.01 (s, 3H), 3.67 (s, 2H), 2.95 - 2.82 (m, 4H), 2.77 -2.70 (m, 2H).Example 948182500867.1

[0163] (6-(4-((4-(lH-Pyrazol-4-yl)phenyl)amino)-7-(2-fluoroethyl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-lH-indol-2-yl)(3,3-difluoropyrrolidin-l-yl)methanone

[0164] Synthesis:

[0165] Step 1: (3,3-Difluoropyrrolidin-l-yl)(6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-IT / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-1 -methyl- 17 / -indol -2 -yl)methanone. To a solution of 6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-17 / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-lJ / -indole-2-carboxylic acid (140 mg, 0.35 mmol) in DMF (2 mL) were added DIEA (91.1 mg, 0.705 mmol), HATU (178.7 mg, 0.47 mmol) and 3,3-difluoropyrrolidine (37.8 mg, 0.352 mmol). The mixture was stirred at 25 °C for 2 hours. The reaction mixture was diluted with ice-water and extracted with EtOAc (3x8 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated, and then purified by column chromatography on silica gel eluting with methanol (from 0 % to 10 %) in dichloromethane to give (3,3-difluoropyrrolidin-l-yl)(6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-17 / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l -methyl- 17 / -indol-2-yl)methanone (115.00 mg, 0.168 mmol, 71.5 % yield) as a yellow solid. LC-MS (ESI): 685.3, [M+H]+.

[0166] Step 2: (6-(4-((4-(17 / -Pyrazol-4-yl)phenyl)amino)-7-(2-fluoroethyl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-177-indol-2-yl)(3,3-difluoropyrrolidin-l-yl)methanone. To a solution of (3,3-difluoropyrrolidin-l-yl)(6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-17 / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4- 49182500867.1d]pyrimidin-2-yl)-l -methyl- U / -indol-2-yl)methanone (115.00 mg, 0.168 mmol) in MeOH (5 mL) was added HC1 (2 mL). The mixture was stirred at 25 °C for 16 hours. The reaction was concentrated and adjusted pH to 9 by aq. NaHCOs and extracted with EtOAc (3×8 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (Waters 3767QDA Column: XBridge C18, 19*250mm, 10 pm; Mobile Phase A: 0.1% NH4OH / H2O, B: ACN; flow rate: 20 mL / min; gradient: 46-46%; Retention Time: 6.7-8.0min of 17min) to afford (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)-7-(2-fluoroethyl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-lJ7-indol-2-yl)(3,3-difluoropyrrolidin-l-yl)methanone (56.94 mg, 0.0948 mmol, 56.4 % yield) as a white solid. LC-MS (ESI): 601.5, [M+H]+.XHNMR (400 MHz, DMSO ) 8 12.89 (s, 1H), 8.52 (s, 1H), 8.44 (s, 1H), 8.17 (s, 1H), 8.11 (dd, J= 8.4, 1.3 Hz, 1H), 7.94 (s, 1H), 7.87 (d, J= 8.7 Hz, 2H), 7.67 (dd, J= 11.6, 8.6 Hz, 3H), 6.98 (s, 1H), 4.74 (t, J= 4.8 Hz, 1H), 4.62 (t, J= 4.8 Hz, 1H), 4.18 (s, 1H), 4.05 - 3.88 (m, 5H), 3.86 - 3.75 (m, 1H), 3.67 (s, 2H), 2.97 - 2.82 (m, 4H), 2.77 - 2.70 (m, 2H), 2.49 - 2.42 (m, 2H).Example 10

[0167] (6-(4-((4-(lH-Pyrazol -4-yl)phenyl)amino)-6-(2-fluoroethyl)-5, 6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lH-indol-2-yl)(3,3-difluoropyrrolidin-l-yl)methanone

[0168] Synthesis:

[0169] Step 1: (3,3-Difluoropyrrolidin-l-yl)(6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-l / / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)- 50182500867.11 -methyl- U / -indol -2 -yl)methanone. To a solution of 6-(6-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2 / / -pyran-2-yl)-l / / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-U / -indole-2-carboxylic acid (200 mg, 0.336 mmol) in DMF (2 mL) were added DIEA (130.1 mg, 1.01 mmol), HATU (255 mg, 0.672 mmol) and 3,3-difluoropyrrolidine (37.8 mg, 0.352 mmol). The mixture was stirred at 25 °C for 2 hours. The reaction mixture was diluted with ice-water and extracted with EtOAc (3x8 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography on silica gel eluting with methanol (from 0% to 10 %) in dichloromethane to give (3,3-difluoropyrrolidin-l-yl)(6-(6-(2-fluoroethyl)-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l -methyl- U / -indol-2-yl)methanone (115.00 mg, 0.168 mmol, 71.5 % yield) as a yellow solid. LC-MS (ESI): 685.3, [M+H]+.

[0170] Step 2: (6-(4-((4-(U / -Pyrazol-4-yl)phenyl)amino)-6-(2-fluoroethyl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-U7-indol-2-yl)(3,3-difluoropyrrolidin-l-yl)methanone. To a solution of (3,3-difluoropyrrolidin-l-yl)(6-(6-(2-fluoroethyl)-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-1H-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-1-methyl-1H-indol-2-yl)methanone (115.00 mg, 0.168 mmol) in MeOH (5 mL) was added HC1 (2 mL). The mixture was stirred at 25 °C for 16 hours. The reaction was concentrated and adjusted pH to 9 by aq. NaHCOs and extracted with EtOAc (3x8 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (Waters 3767QDA Column: XBridge C18, 19*250mm, lO um; Mobile Phase A: 0.1% NH4OH / H2O, B: ACN; flow rate: 20 mL / min; gradient: 46-46%; Retention Time: 6.7-8.0min of 17min) to afford (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)-6-(2-fluoroethyl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-1-methyl-1H-indol-2-yl)(3,3-difluoropyrrolidin-1-yl)methanone (32.30 mg, 0.0538 mmol, 32.0 % yield) as a white solid. LC-MS (ESI): 601.5, [M+H]+.XHNMR (400 MHz, DMSO-d6) 5 13.12 - 12.68 (m, 1H), 8.45 (s, 1H), 8.41 (s, 1H), 8.12 (d, J= 8.4 Hz, 3H), 7.82 (d, J= 8.6 Hz, 2H), 7.67 (t, J= 8.5 Hz, 3H), 6.98 (s, 1H), 4.71 (dt, J=47.5, 4.9 Hz, 2H), 4.18 (s, 1H), 3.93 (d, J= 16.1 Hz, 5H), 3.80 (s, 1H), 3.65 (s, 2H), 2.97 (t, J= 5.1 Hz, 1H), 2.94 - 2.83 (m, 5H), 2.49 - 2.40 (m, 2H).Example 1151182500867.1

[0171] (6-(4-((4-(lH-Pyrazol-4-yl)phenyl)amino)-6-(2-fluoroethyl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone

[0172] Synthesis:THP

[0173] Step 1: Methyl 6-(6-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-lJT-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-l / / -indole-2-carboxylate. To a solution of methyl l-methyl-6-(4-((4-(l-(tetrahydro-2 / / -pyran-2-yl)-l / / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l / / -indole-2-carboxylate (1 g, 1.77 mmol) and l-bromo-2-fluoroethane (450 mg, 3.55 mmol) in A A -di N,N-dimethylformamide (20 mL) was added K2CO3 (734 mg, 5.32 mmol). The mixture was stirred at 80 °C for 2 hrs. The reaction mixture was diluted with ice-water and extracted with EA (3x50 mL) and the combined organic layers were dried over Na₂SO₄ and concentrated, then purified by column chromatography on silica gel eluting with methanol (from 0% to 10 %) in di chloromethane to afford methyl 6-(6-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2 / / -pyran-2-yl)-17 / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-52182500867.1methyl-U / -indole-2-carboxylate (630 mg, 1.03 mmol, 58.2 % yield) as a yellow solid. LC-MS (ESI): 610.2, [M+H]+.

[0174] Step 2: 6-(6-(2-Fluoroethyl)-4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-lJff-pyrazol-4-yl)phenyl)amino)-5, 6,7, 8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l -methyl- lJ / -indole-2-carboxylic acid. To a solution of methyl 6-(6-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-17 / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-lJT-indole-2-carboxylate (680 mg, 1.12 mmol) in methanol (10 mL) and THF (10 mL) and H2O (3 mL) were added LiOH.H2O (234 mg, 5.58 mmol). The mixture was stirred at room temperature for 12 hrs. The reaction was adjusted pH to 3~4, and extracted with EA (3x50 mL) and the combined organic layers were dried over Na2SO4 and concentrated to afford 6-(6-(2-fluoroethyl)-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5, 6,7, 8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l -methyl- U / -indole-2-carboxylic acid (580 mg, 0.973 mmol, 86.9 % yield) as a yellow solid. LC-MS (ESI): 596.3, [M+H]+.

[0175] Step 3: (3,3-Difluoroazetidin-l-yl)(6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2JH-pyran-2-yl)-lJ / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-1 -methyl- UT-indol -2 -yl)methanone. To a solution of 6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-lJ / -indole-2-carboxylic acid (150 mg, 0.252 mmol) in DMF (2 mL) were added DIEA (97.5 mg, 0.756 mmol), HATU (192 mg, 0.504 mmol) and 3,3-difluoroazetidine (48.9 mg, 0.378 mmol). The mixture was stirred at 25 °C for 2 hours. The reaction mixture was diluted with ice-water and extracted with EtOAc (3x8 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography on silica gel eluting with methanol (from 0% to 10 %) in dichloromethane to give (3,3-difluoroazetidin-l-yl)(6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l -methyl- UT-indol -2 -yl)methanone (156.00 mg, 0.232 mmol, 92.3 % yield) as a yellow solid. LC-MS (ESI): 671.2, [M+H]+.

[0176] Step 4: (6-(4-((4-(17 / -Pyrazol-4-yl)phenyl)amino)-6-(2-fluoroethyl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-l-methyl-U7-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone. To a solution of (3,3-difluoroazetidin-l-yl)(6-(7-(2-fluoroethyl)-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l -methyl- 17 / -indol-2-yl)methanone (156.00 mg, 0.232 mmol) in MeOH (5 mL) was added HC1 (2 mL). The mixture was stirred at 25 °C for 16 hours. The reaction was 53182500867.1concentrated and adjusted pH to 9 by aq. NaHCOs and extracted with EtOAc (3×8 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (Waters 3767QDA Column: XBridge C18, 19*250mm, 10 pm; Mobile Phase A: 0.1% NH4OH / H2O, B: ACN; flow rate: 20 mL / min; gradient: 46-46%; Retention Time: 6.5-8.0min of 17min) to afford (6-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)-6-(2-fluoroethyl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-2-yl)-1-methyl-1H-indol-2-yl)(3,3-difluoroazetidin-1-yl)methanone (67.27 mg, 0.115 mmol, 49.4 % yield) as a white solid. LC-MS (ESI): 587.4, [M+H]+.XHNMR (400 MHz, DMSO-d6) 5 12.91 (s, 1H), 8.47 (s, 1H), 8.42 (s, 1H), 8.13 (dd, J= 8.5, 1.3 Hz, 2H), 7.95 (s, 1H), 7.81 (d, J= 8.7 Hz, 2H), 7.67 (dd, J= 10.4, 8.7 Hz, 3H), 4.71 (dt, J= 47.6, 5.0 Hz, 6H), 4.03 (d, J= 16.0 Hz, 3H), 3.65 (s, 2H), 3.02 - 2.84 (m, 6H).Example 12

[0177] (6-(4-((4-(lH-Pyrazol -4-yl)phenyl)amino)-7-cy cl opropyl-5, 6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-lH-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone

[0178] Synthesis:

[0179] Step 1: (6-(7-Cyclopropyl-4-((4-(l-(tetrahydro-2 / / -pyran-2-yl)-l / / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-l / / -indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone. To a solution of (3,3-difluoroazetidin-l-yl)(l-methyl-6-(4-((4-(l-(tetrahydro-2J / -pyran-2-yl)-U / -pyrazol-4-yl)phenyl)amino)-5, 6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-1H-indol-2-yl)methanone (270 mg, 0.433mmol) in 54182500867.1DCE (3 mL) were added cyclopropylboronic acid (112 mg, 1.299 mmol), 4,4'-bipyridine (68 mg, 0.433 mmol), Cu(OAc)2 (79 mg, 0.433 mmol) and Na2COs (138 mg, 1.299 mmol).Reaction mixture was purged with O2 for three times. The mixture was stirred at 60 °C overnight under O2 atmosphere. The reaction mixture was diluted with water and extracted with EtOAc (3x10 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography on silica gel eluting with methanol (from 0% to 15 %) in di chloromethane to give (6-(7-cyclopropyl-4-((4-(l-(tetrahydro-27 / -pyran-2-yl)-lJH-pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-U / -indol-2-yl)(3,3-difluoroazetidin-l-yl)m ethanone (160.00 mg, 0.241 mmol, 55.6 % yield) as a yellow solid. LC-MS (ESI): 665.3, [M+H]+.

[0180] Step 2: (6-(4-((4-(U / -Pyrazol-4-yl)phenyl)amino)-7-cyclopropyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l-methyl-177-indol-2-yl)(3,3-difluoroazetidin-l-yl)methanone. To a solution of (6-(7-cyclopropyl-4-((4-( l-(tetrahydro-27 / -pyran-2-yl)- IT / -pyrazol-4-yl)phenyl)amino)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l -methyl- 1H-indol-2-yl)(3,3-difluoroazetidin-1-yl)methanone (160.00 mg, 0.241 mmol) in MeOH (5 mL) was added HC1 (2 mL). The mixture was stirred at 25 °C for 6 hours. The reaction was concentrated and adjusted pH to 9 by aq. NaHCOs and extracted with EtOAc (3x10 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (Waters 3767QDA Column: XBridge C18, 19*250mm, 10 pm; Mobile Phase A: 0.1% NH4HCO3 / H2O, B: ACN; flow rate: 20 mL / min; gradient: 46-46%; Retention Time: 6.7-8.0 min of 17min) to afford (6-(4-((4-(U / -pyrazol-4-yl)phenyl)amino)-7-cyclopropyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)-l -methyl- 177-indol -2 -yl)(3, 3 -difluoroazeti din- 1-yl)methanone (26.50 mg, 0.0456 mmol, 30.31 % yield) as a white solid. LC-MS (ESI): 581.6, [M+H]+.XHNMR (400 MHz, DMSO-d6) 5 12.89 (s, 1H), 8.48 (d, J= 8.9 Hz, 2H), 8.18 -7.93 (m, 3H), 7.85 (d, J= 8.6 Hz, 2H), 7.67 (dd, J= 15.9, 8.5 Hz, 3H), 7.04 (s, 1H), 5.14 -4.24 (m, 4H), 4.01 (s, 3H), 3.72 (s, 2H), 3.02 - 2.94 (m, 2H), 2.73 - 2.65 (m, 2H), 1.93 - 1.85 (m, 1H), 0.58 - 0.50 (m, 2H), 0.49 - 0.40 (m, 2H).Example 13

[0181] ROCK2 Inhibition

[0182] The inhibition of ROCK 1 and ROCK 2 was measured using an ADP-Glo kinase assay (Promega). The ADP-Glo™ Kinase Assay is a luminescent ADP detection assay that allows measurement of kinase activity based on the amount of ADP produced during a kinase 55182500867.1reaction. The kinase reaction is performed in the presence of ATP, S6K substrate (KRRRLASLR) and the ROCK kinase in the appropriate Kinase Reaction Buffer (lx). Upon termination of the reaction, the unconsumed ATP is depleted by addition of ADP-Glo™ Reagent. Kinase Detection Reagent is added to convert ADP to ATP and allow the newly synthesized ATP to be measured using a luciferase / luciferin reaction. The luminescence is proportional to the produced ADP and consequently to the kinase activity.Table 1Comp. # ROCK2 IC50 ROCK1 IC50(nM) (nM)Ex. 1 5.741 >100000Ex. 2 4.59 4623Ex. 3 2.712 2321Ex. 4 240.2 >100000Ex. 5 58.61 >100000Ex. 6 4.52 33046Ex. 7 12.82 >100000Ex. 8 19.17 >100000Ex. 9 14.66 41782Ex. 10 20 10023Ex. 11 9.56 14866Ex. 12 137.06 >100000Example 14

[0183] Permeability and Efflux Ratio Determination of Compound 1 (Example 1) in Caco-2 Cells.

[0184] Caco-2 cell Culture: Caco-2 were seeded in 96-transwell plate insert systems at 1 x 105 cells / cm2 until to 14-28 days for confluent cell monolayer formation. Medium was changed every 3-4 days.

[0185] Experimental Procedures: Test compounds were diluted with the transport buffer (HBSS with BSA) from a 10 mM stock solution to a concentration 10 pM and applied to the apical or basolateral side of the cell monolayer. Permeation of the test compounds from A to B direction or B to A direction was determined in duplicate over a 120 minute incubation at 37°C and 5% CO2 with a relative humidity of 95%. In addition, the efflux ratio of each compound was also determined. Test and reference compounds were quantified by LC-MS / MS analysis based on the peak area ratio of analyte / IS.56182500867.1

[0186] Data Analysis: The apparent permeability coefficient Papp (cm / s) was calculated using the equation:Papp = (dCr / dt) * Vr / (A * CO)

[0187] Where dCr / dt is the cumulative concentration of compound in the receiver chamber as a function of time (S); Vr is the solution volume in the receiver chamber (0.1 mL on the apical side, 0.3 mL on the basolateral side); A is the surface area for the transport, i.e.0.143 cm2 for the area of the monolayer; CO is the initial concentration in the donor chamber.

[0188] The efflux ratio was calculated using the equation:Efflux Ratio = Papp (BA) / Papp (AB)

[0189] Percent recovery was calculated using the equation:% Recovery = 100 * [(Vr x Cr) + (Vd x Cd)] / (Vd x CO)% Total recovery =100 x [(Vr x Cr) + (Vd x Cd) + (Vc x Cc)] / (Vd x CO)

[0190] Where Vd is the volume in the donor chambers (0.1 mL on the apical side, 0.3 mL on the basolateral side); Cd and Cr are the final concentrations of transport compound in donor and receiver chambers, respectively. Cc is the compound concentration in the cell lysate solution. Vc is the volume of insert well (0.1 mL in this assay).Table 2. LC / MS / MS Conditions.DetectionShimadzu NexeraLC-30AD & SCIEX TQ-6500+methodMatrix HBSSInternalTolbutamide / Verapamilstandard (s)MS ESI Positive-ion for Atenolol, Proranolol & Comp. 1conditions ESI Negative-ion for DigoxinMobile A: 0.1%FA in H2O B: 0.1%FA in ACNphaseColumn ACQUITY UPLC HSS T3( 1.8 pm, 2.1x50 mm)For Digoxin+Atenolol+Propranolol For Comp. 1 0.6 mL / min 0.6 ml / min Time Pump B Time Pump B 0 5 0 10 LC 0.2 5 0.5 95 conditions 0.7 95 1 951.2 95 1.01 10 1.21 5 1.5 10 1.7 5

[0191] The results are summarized in Table 3 below:57182500867.1Table 3. Results Summary for Permeability and Efflux Ratio Determination of Compound 1 in Caco-2 Cells.Summary Papp (10-6 cm / s) Efflux Ratio Recovery % Compound ID A to B B to A Ato B B to A A to B B to A Atenolol 0.32 NA 100.4 NAPropranolol 24.63 NA 106.2 NADigoxin 0.31 16.38 53.05 99.9 95.8Comp. 1 0.32 1.86 5.92 75.2 92.9 93.7 94.1Example 15

[0192] The Nano BRET® TE Intracellular Kinase Assays can analyze the affinity of test compounds by competitive displacement of a fluorescent Nano BRET® Tracer reversibly bound to a Nano Luc®-kinase fusion protein in cells. Introduction of competing test compounds results in a dose-dependent decrease in the Nano BRET® signal for estimating intracellular affinity against the target protein.

[0193] The objective of this study is to quantitatively measure the inhibition from Compound 1 in ROCK1 and ROCK2 expressed in HEK293 live cells by Nano BRET readout at Pharmaron.

[0194] The NanoBRET® Target Engagement (TE) Intracellular Kinase Assays were used to quantitatively measure the inhibition of Compound 1 on human ROCK1 and Human ROCK2 kinase in vivo.

[0195] The Nano BRET® Assays analyze the affinity of test compounds by competitive displacement of a fluorescent Nano BRET® Tracer reversibly bound to a Nano Luc®-ROCK kinase fusion protein in cells. Introduction of competing test compound, here Compound 1, results in a dose-dependent decrease in the Nano BRET® signal for estimating intracellular affinity against the target ROCK1 or ROCK2 protein.

[0196] Before testing the compound, the tracer was optimized with a tracer concentration gradient of 2pM, 2x dilution, 11 points, to find the appropriate concentration to test compound, and select the tracer concentration of 0.5pM and 0.25 pM test compounds. Results showed that for the inhibition from Compound 1 on human ROCK1 kinase, the half-maximal inhibitory concentration was not reached with the highest concentration of 10 pM; but for human ROCK2 kinase, the IC50 is 79.04nM with Trace 16 concentration 0.5 pM and 64.8nM with Trace 16 at 0.25 pM.58182500867.1

[0197] For the concentration range we measured (0.17nM to lOOOOnM), Compound 1 showed high selective inhibition on ROCK2 (IC5064.8 / 79.04nM) compared to ROCK1, which did not reach the IC50 with the highest concentration (lOOOOnM).Methods

[0198] Cell seeding: HEK293 cells were cultivated in T-75 flasks in a cell culture incubator set at 37°C, 5% CO2, 95% relative humidity. Allow cells to reach 80-90% confluence before detaching and splitting. Remove medium from cell flask by aspiration, trypsinize and allow cells to dissociate from the flask. Neutralize trypsin using Cell Culture Medium and centrifuge at 200 * g for 5 minutes to pellet cells. Aspirate medium and resuspend cells in Assay Medium. Adjust density to 2 * 105 cells / mL using Assay Medium. Prepare a lOpg / mL solution of DNA in Opti-MEM® I Reduced Serum Medium, no phenol red that consists of the following ratios: 9.0pg / mL of Transfection Carrier DNA, LOpg / mL of Nano Luc® ROCK1 / Nano Luc® ROCK2 and ImL of Opti-MEM® I. mix thoroughly. Add 30pL of Fu GENE® HD Transfection Reagent into each milliliter of DNA mixture to form lipid: DNA complex. Mix by inversion 5-10 times. Incubate at ambient temperature for 20 minutes to allow complexes to form. In a sterile, conical tube, mix 1 part of lipid: DNA complex (e.g. ImL) with 20 parts of HEK293 cells (e.g. 20mL) in suspension at 2 * 105 cells / mL. Mix gently by inversion 5 times. Dispense 40pL cells + lipid: DNA complex into a sterile tissue-culture treated 384-well assay plate and incubate 20-30 hours.

[0199] Tracer optimization: Transfer 40 pL of 400 pM stock solution to a 384 pp-plate, perform 2-fold series dilution, 11 points, transferring 20 pL tracer into 20 pL tracer dilution buffer. Incubate the plate at 37°C, 5% CO2 for 2 hours.

[0200] Compound preparation and treatment: Compounds are dissolved in DMSO to make 10 mM stock solution. Transfer 40 pL of 10 mM stock solution to a 384 pp-plate (LABCYTE, PP-0200). Perform 3-fold, 10-point dilution via transferring 12 pL compound into 24 pL DMSO by Apricot. Transfer 40 nL / well the serial dilution of test compound to 384-well solid white plate by Echo; the DMSO concentration is 0.1% and consistent in all wells. The final tracer 16 concentration is 0.5uM: Prepare 400 pM Nano BRET™ Tracer Reagent. Transfer 50 nL / well tracer 16 to 384 well plate by ECHO 655. The final tracer concentration is 0.5 pM. The final tracer 16 concentration is 0.25uM: Dilute 400 pM Nano BRET™ Tracer Reagent to 200 pM. Transfer 50 nL / well 200 pM tracer 16 to 384 well plate by ECHO 655. Mix cell plates on an orbital shaker for 15 seconds at 700 rpm. Incubate the plate at 37°C, 5% CO2 for 2 hours.59182500867.1

[0201] Detection: Prepare 3X Complete Substrate in Opti-MEM® I Reduced Serum Medium, no phenol red as flows: Nano BRET™ Nano-Gio® Substrate 48uL; Extracellular Nano Luc® Inhibitor 16uL; Opti-MEM® reduced serum medium, no phenol red 7936uL. Add 20pl of 3X Complete Substrate plus Inhibitor Solution to each well of the 384-well plate. Incubate for 2-3 minutes at room temperature. Read the plate on the Envision.

[0202] Data analysis: BRET Ratio mBU = Acceptor sample / Donor sample×1000

[0203] GraphPad Prism 8.0.2 (263) protocol non-linear regression (curve fit), Dose-response, log(inhibitor) vs. response- variable slope (four parameters).

[0204] Positive control: Cells+DMSO with tracer.

[0205] Negative control: Cells+DMSO without tracer.

[0206] Results: The tracer optimization curves of ROCK1 and ROCK2 expressed in HEK293 cells by Nano BRET is shown in FIG. 1. Dose response curves of Compound 1 in ROCK1 and ROCK2 expressed in HEK293 cells by Nano BRET is shown in FIG. 2. A summary of the IC50 data of Compound 1 in ROCK1 and ROCK2 is shown in Table 4. Table 4. IC50 Summary of Compound 1 in ROCK1 and ROCK2.Trace 16 Conc.(iiM) Cell Compound ID ROCK1 IC50 (nM) ROCK2 IC50 (nM) 0.5 HEK293 Comp. 1 NA 79.040.25 HEK293 Comp. 1 NA 64.80

[0207] Conclusion: For the concentration range measured (0.17nM to lOOOOnM), Compound 1 showed high selective inhibition on ROCK2 (IC5064.8 / 79.04nM) compared to ROCK1, which did not reach the IC50 with the highest concentration (lOOOOnM).182500867.1

Claims

We Claim:

1. A compound having the Formula I:iR3jor a pharmaceutically acceptable salt thereof, wherein:X1is selected from the group consisting of CH2, O and NR4;X2is selected from the group consisting of CH2, O and NR4;wherein one of X1or X2is CH2; and one of X1or X2is O or NR4;R1is selected from the group consisting of H, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3- C7 cycloalkyl, C5-C10 aryl, C5-C10 heteroaryl, 3- to 10-membered heterocyclyl, -Ci-Ce alkyl-(C3-C? cycloalkyl), -Ci-Ce alkyl-(C5-Cio aryl), -Ci-Ce alkyl-(C5-Cio heteroaryl), - Ci-Ce alkyl-(3- to 10-membered heterocyclyl), -(Ci-Ce alkyl)-NRnR12, -(Ci-Ce alkyl)- OR11, and -C(=O)-N(R11)(R12), and wherein each alkyl, alkenyl, cycloalkyl, aryl, heteroaryl and heterocyclyl may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;each R11and R12are independently selected from the group consisting of H and Ci-Ce alkyl;or alternatively, R11and R12are taken together when both are attached to the same nitrogen to form a 4- to 7- membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from the group consisting of N, O and S, and which heterocyclic ring is unsubstituted or is substituted with 1 to 3 substituents selected from the group consisting of halo, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, -CN, -NH2, C1-C3 perfluoroalkyl, -OH, -O-(Ci-Ce alkyl), and -(Ci-Ce alkyl)-OH;R2and R3are each independently selected from the group consisting of H, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, 3- to 10-membered heterocyclyl, Ce-Cio aryl, 5- to 14-membered heteroaryl, wherein each alkyl, alkenyl, cycloalkyl, heterocyclyl, aryl,61182500867.1and heteroaryl may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;alternatively R2and R3are taken together with the nitrogen to which they are attached to provide (i) a 4- to 6-membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from N, O and S, or (ii) a 5- to 10-membered hetero bicyclic ring system having from 0 to 3 additional ring heteroatoms selected from N, O and S; wherein the heterocyclic ring or the bicyclic ring system are unsubstituted or are substituted with from 1 to 4 substituents selected from the group consisting of halo, Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, -CN, Ci-Ce fluoroalkyl, C1-C3 perfluoro alkyl, - OR11, oxo, and -NH2; andR4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

2. The compound of claim 1, having the Formula IA:or a pharmaceutically acceptable salt thereof, wherein:X1is selected from the group consisting of CH2, O and NR4;X2is selected from the group consisting of CH2, O and NR4;wherein one of X1or X2is CH2; and one of X1or X2is O or NR4;R1Ais selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;R2Aand R3Aare each independently selected from the group consisting of H, Ci-Ce alkyl, C2- Ce alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, wherein each alkyl, alkenyl, cycloalkyl, may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;62182500867.1alternatively R2Aand R3Aare taken together with the nitrogen to which they are attached to provide a 4- to 6-membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from N, O and S, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of halo, C1-C3 alkyl, -CN, C1-C3 fluoroalkyl, C1-C3 perfluoro alkyl, -NH2, -OH, and -O-C1-C3 alkyl; andR4is selected from the group consisting of H, Ci-Ce alkyl and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

3. The compound of claim 1, having the Formula IB:(R > IBor a pharmaceutically acceptable salt thereof, wherein:X1is selected from the group consisting of CH2, O and NR4;X2is selected from the group consisting of CH2, O and NR4;wherein one of X1or X2is CH2; and one of X1or X2is O or NR4;R1Bis selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;each R5is independently selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;n is 1 or 2;m is 0, 1 or 2; andR4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.63182500867.

14. The compound of claim 1, having the Formula IIA:or a pharmaceutically acceptable salt thereof, wherein:X1is selected from the group consisting of O and NR4;R1Ais selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;R2Aand R3Aare each independently selected from the group consisting of H, Ci-Ce alkyl, C2- Ce alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, wherein each alkyl, alkenyl, cycloalkyl, may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;alternatively R2Aand R3Aare taken together with the nitrogen to which they are attached to provide a 4- to 6-membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from N, O and S, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of halo, C1-C3 alkyl, -CN, C1-C3 fluoroalkyl, C1-C3 perfluoro alkyl, -NH2, -OH, and -O-C1-C3 alkyl; and R4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

5. The compound of claim 1, having the Formula IIIA:182500867.1or a pharmaceutically acceptable salt thereof, wherein:R1Ais selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;R2Aand R3Aare each independently selected from the group consisting of H, Ci-Ce alkyl, C2- Ce alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, wherein each alkyl, alkenyl, cycloalkyl, may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2; andalternatively R2Aand R3Aare taken together with the nitrogen to which they are attached to provide a 4- to 6-membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from N, O and S, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of halo, C1-C3 alkyl, -CN, C1-C3 fluoroalkyl, C1-C3 perfluoro alkyl, -NH2, -OH, and -O-C1-C3 alkyl.

6. The compound of claim 1, having the Formula IIIB:or a pharmaceutically acceptable salt thereof, wherein:65182500867.1R1Bis selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;each R5is independently selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;n is 1 or 2; andm is 0, 1 or 2.

7. The compound of claim 1, having the Formula IVA:HN N N R1AI NN'R3A / IVA or a pharmaceutically acceptable salt thereof, wherein:R1Ais selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;R2Aand R3Aare each independently selected from the group consisting of H, Ci-Ce alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C7 cycloalkyl, wherein each alkyl, alkenyl, cycloalkyl, may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;alternatively R2Aand R3Aare taken together with the nitrogen to which they are attached to provide a 4- to 6-membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from N, O and S, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of halo, C1-C3 alkyl, -CN, C1-C3 fluoroalkyl, C1-C3 perfluoro alkyl, -NH2, -OH, and -O-C1-C3 alkyl; andR4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.182500867.

18. The compound of claim 1, having the Formula IVB:or a pharmaceutically acceptable salt thereof, wherein:R1Bis selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;each R5is independently selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;n is 1 or 2;m is 0, 1 or 2; andR4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

9. The compound of claim 1, having the Formula VA:or a pharmaceutically acceptable salt thereof, wherein:R1Ais selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;67182500867.1R2Aand R3Aare each independently selected from the group consisting of H, Ci-Ce alkyl, C2- Ce alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, wherein each alkyl, alkenyl, cycloalkyl, may optionally be substituted with one to three substituents selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;alternatively R2Aand R3Aare taken together with the nitrogen to which they are attached to provide a 4- to 6-membered heterocyclic ring having from 0 to 2 additional ring heteroatoms selected from N, O and S, wherein the heterocyclic ring is unsubstituted or is substituted with from 1 to 4 substituents selected from the group consisting of halo, C1-C3 alkyl, -CN, C1-C3 fluoroalkyl, C1-C3 perfluoro alkyl, -NH2, -OH, and -O-C1-C3 alkyl; andR4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

10. The compound of claim 1, having the Formula VB:HN NNor a pharmaceutically acceptable salt thereof, wherein:R1Bis selected from the group consisting of H and Ci-Ce alkyl, optionally substituted with one to three substituents selected oxo, halo, -CN, -OH, and NH2;each R5is independently selected from C1-C3 alkyl, C1-C3 perfluoroalkyl, oxo, halo, -CN, -OH, and NH2;n is 1 or 2;m is 0, 1 or 2; and182500867.1R4is selected from the group consisting of H, Ci-Ce alkyl, and C3-C6 cycloalkyl, wherein the Ci-Ce alkyl and C3-C6 cycloalkyl are optionally substituted by 1 to 3 substituents selected from halo, hydroxy, and C1.3 alkoxy.

11. The compound of claim 1, having a formula selected from:69182500867.1or a pharmaceutically acceptable salt thereof.

12. The compound of claim 1, which is (6-(4-((4-(lH-pyrazol-4-yl)phenyl)amino)-7,8-dihydro-5H-pyrano[4, 3-d]pyrimidin-2-yl)-l -methyl- lH-indol-2-yl)(3,3-difluoroazeti din- 1-yl)methanone, having the formula:or a pharmaceutically acceptable salt thereof.

13. A method for the treatment of a disease or disorder mediated by R0CK2, wherein the method comprises administering to a subject in need thereof an effective amount of a compound according to any of claims 1 to 12 or a pharmaceutically acceptable salt thereof.

14. The method of claim 13, wherein the disease or disorder is selected from the group consisting of fibrotic diseases, inflammatory diseases, autoimmune diseases, cardiovascular 70182500867.1disorders, central nervous system disorders, neoplastic diseases, metabolic syndromes, ocular diseases, renal diseases, pulmonary diseases, muscular dystrophy, sickle cell disease, and viral diseases.

15. The method of claim 14, wherein the disease or disorder is selected from the group consisting of fibrotic diseases, inflammatory diseases, and autoimmune diseases.

16. The method of claim 15, wherein autoimmune disease is selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus (SLE; lupus), psoriasis, psoriatic arthritis, multiple sclerosis, Crohn’s disease, ulcerative colitis, atopic dermatitis, eczema, or graft-versus-host disease (GVHD; acute and chronic), idiopathic pulmonary fibrosis and scleroderma.

17. The method of claim 14, wherein the disease or disorder is selected from the group consisting of a cardiovascular disorder, a central nervous system disorder, a neoplastic disease, or a metabolic syndrome.

18. The method of claim 15, wherein the inflammatory disorder is selected from the group consisting of cardiovascular inflammation, pulmonary inflammation, renal inflammation, arteriosclerosis and sepsis.

19. The method of claim 15, wherein the fibrotic disorder is selected from the group consisting of idiopathic pulmonary fibrosis, renal fibrosis, kidney fibrosis, ocular fibrosis, cardiac fibrosis, NASH, scleroderma, systemic sclerosis, and cirrhosis.

20. The method of claim 17, wherein the neoplastic disease is selected from the group consisting of ovarian cancer, breast cancer and pancreatic cancer.

21. The method of claim 17, wherein the cardiovascular disease is selected from the group consisting of hypertension, cardiomyopathy, cardiac remodeling, atherosclerosis, restenosis, cardiac hypertrophy, cerebral ischemia, cerebral vasospasm, and erectile dysfunction.71182500867.

122. The method of claim 14, wherein the pulmonary disease is selected from the group consisting of idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, and asthma.

23. The method of claim 17, wherein the central nervous system disorder is selected from the group consisting of neuronal degeneration or spinal cord injury, traumatic brain injury, cerebral cavernous malformation, Huntington’s disease, Parkinson’s disease, Alzheimer’s disease, Amyotrophic lateral sclerosis (ALS), and multiple sclerosis.

24. The method of claim 14, wherein the renal disease is selected from the group consisting of polycystic kidney disease, renal fibrosis and diabetic renal disease.

25. The method of claim 14, wherein the metabolic disease is selected from the group consisting of insulin resistance, hyperinsulinemia, type 2 diabetes, obesity, metabolic syndrome and glucose intolerance.

26. The method of claim 14, wherein the ocular disease is selected from the group consisting of ocular hypertension, age related macular degeneration (AMD; wet and dry), choroidal neovascularization (CNV), choroidal tumor, diabetic macular edema (DME), iris neovascularization, uveitis, glaucoma, primary open-angle glaucoma, acute angle-closure glaucoma, pigmentary glaucoma, congenital glaucoma, normal tension glaucoma, secondary glaucoma, neo vascular glaucoma, geographic atrophy, and retinitis of prematurity (ROP).

27. The method of claim 13, wherein the disease is Duchenne muscular dystrophy.

28. The method of claim 14, wherein the viral infection is a coronavirus infection such as SARS-CoV-1, SARS-CoV-2, and MERS-CoV.72182500867.1

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