HETEROCYCLIC COMPOUNDS AS ERK INHIBITORS

Field of the Invention

[01] The present application is in general in the field of medicinal chemistry and specifically related to compounds that are protein kinase inhibitors, such as inhibitors of extracellular signal- regulated kinases (also known as "ERK"). These compounds are useful for treating disease and conditions (e.g., cancers) that are associated with such protein kinases as ERK.

Background of the Invention

[02] The Ras-Raf-Mek-Erk intracellular signaling cascade is a central signaling module that transmits proliferation, survival, growth and differentiation signals into the cell interior from the activated receptor tyrosine kinases (RTKs) such as ErbB family, PDGF, FGF, and VEGF (Sebolt-Leopold, J. S. and Herrera, R., Nat. Rev. Cancer, 41 :937-947, 2004; Kolch, W., Nat. Rev. Mol. Cell Biol, 61 :827-837, 2005). This signaling axis is comprised of Ras, Raf, Mek (mitogen-activated protein kinase kinase), and Erk (extracellular signal-regulated kinases) proteins all occurring in highly homologous isoforms. Ras proteins (H, N, and K) are 21 kDa GTPases that are activated at the proximity sites of the intracellular kinase domains of RTKs. Raf kinases (A, B, and C) are intermediate downstream effectors of Ras, activated by binding to GTP-loaded Ras. Raf kinases phosphorylate Meks (Mekl and Mek2) on two closely adjacent serine residues, S218 and S222 in the case of Mekl. Meks are dual specificity theroine/tyrosine kinases that phosphorylate threonine and tyrosine residues within the TXY motif of Erks, where X represents any amino acid. Erk proteins (Erkl and Erk2), also known as MAPKs (mitogen- activated proteins), are serine/threonine kinases that phosphorylate more than 100 downstream cytosolic and nuclear target proteins that participate in cellular processes such as division, proliferation, migration, and apoptosis (Yoon, S. and Seger, R., Growth Factors, 24:21-44, 2006). These phosphorylations substantially modulate, generally stimulate, the activity of the target proteins and profoundly alter the physiological status of the cells.

[03] Pathological activation of Ras-Raf-Mek-Erk cascade signaling pathway accounts for the mechanistic aspects of most human cancers, immune dysfunction, and hyper-inflammatory conditions. Activation of the signaling pathway can occur as the result of autocrine or paracrine production of excessive RTK ligands, or constitutive activation of cell surface receptors by mutation or overexpression, or more commonly through gain-of- function mutations of B-Raf and Ras family members. Oncogenic forms of Ras are associated with 30% of all human cancers. Mutations in K-ras occur in 90% of pancreatic and in 25% to 50% of colorectal, mucinous ovarian, and non-small cell lung cancers, whereas mutations in H-ras are common in bladder, kidney, and thyroid cancers and N-ras mutations are found in melanoma, hepatocellular carcinoma, and hematologic malignancies (Adjei, A., J. Nat'l. Cancer Inst., 93 : 1062-74, 2001 ; Aviel-Ronen, S., et al, Clin Lung Cancer, 8:30-8, 2006). B-Raf mutations occur in 66% to 70% of malignant melanomas, 70% of nonpapillary thyroid cancers, 35% of low-grade ovarian serous tumors as well as a wide range of other cancers including colorectal, thyroid, lung, breast, and ovarian (Thomas, N., Melanoma Res, 16:97-103, 2006; Singer, G., et al, J Natl Cancer Inst, 95:484-6, 2003; Brose, M., et al, Cancer Res., 62:6997-7000, 2002).

[04] Inhibition of the activity of Ras-Raf-Mek-Erk signaling pathway has been the focus of drug discovery, particularly for cancer treatment (Sebolt-Leopold, J., Oncogene, 19: 16564-6599, 2000). Small molecules inhibitors of B-Raf and Mek have been shown to effectively inhibit Ras and Raf mediated cell transformation, Erk activation and dependent processes, cell proliferation in vitro, tumor growth in vivo (Mallon, R., et al., Mol. Cancer Ther., 31 :755-762, 2004; Sebolt- Leopold, J., Curr. Pharm Des, 101 : 1907-1914, 2004; Sebolt-Leopold J. and Herrera, R., Nat Rev Cancer, 41 :937-947, 2004). The demonstration of the clinical efficacy of multiple Raf and Mek small molecule inhibitors in various types of cancers has provided an ultimate validation of targeting this signaling pathway for cancer treatment (Crane, E. and Wang, K., Topics Anti- Cancer Res., 2:63-94, 2013).

[05] Given ERK proteins' downstream position in the Ras-Raf-Mek signaling cascade, inhibition of ERK can provide an alternative strategy to modulate down the activity of the pathway. As such, there is a strong rationale to develop ERK small molecule inhibitors as novel therapeutic agents for a broad spectrum of human cancers originated from brain, lung, colon, breast, gastric, pancreatic, head and neck, esophageal, renal, kidney, ovarian, skin, prostate, testicular, gynecological or thyroid. In addition, the ERK inhibitors may also be used to treat non-cancerous hyper-proliferative disorders (e.g., benign hyperplasia of the skin, restenosis, benign prostatic hypertrophy), pancreatitis, kidney disease, pain, diseases related to

vasculogenesis or angiogenesis, acute and chronic inflammatory disease (e.g., rheumatoid arthritis, athero sclerosis, inflammatory bowel disease), skin diseases (e.g., psoriasis, eczema, and scleroderma), diabetes, diabetic retinopathy, retinopathy of prematurity, age-related macular degeneration, asthma, septic shock, T-cell mediated diseases, chronic obstructive pulmonary disease (COPD) and other diseases.

Summary of the Invention

[06] The present invention in general provides compounds that exhibit surprisingly superior effect on inhibiting ERK, pharmaceutical compositions containing the compounds and uses thereof.

[07] In one aspect, the present invention provides compounds or pharmaceutically acceptable salts thereof that, among others, are surprisingly effective ERK inhibitors. These compounds are of Formula I as shown below:

I

In Formula I,

A is a fused 6,5- or 5,6-bicyclic heteroaryl with one ring bonded to B and the other ring not bonded to B, and the ring not bonded to B must have a nitrogen atom and is optionally substituted with one or more Ri groups;

B is a 5-membered heteroaryl having 1 to 3 heteroatoms each of which independently is O, S, or N; and B is optionally substituted with one or more R ₂ groups;

each optional Ri group independently is halogen, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heteroaryl, heterocyclyl, or NRaRb, and when applicable Ri is optionally substituted with 1 to 4 Rc groups;

each of Ra and Rb independently is hydrogen, alkyl, aryl, or heteroaryl, and when applicable is optionally substituted with halogen, hydroxyl, amino, or cyano; or Ra and Rb, together with the nitrogen atom to which they are attached, form an optionally substituted 3- to 7-membered heterocyclic ring with 0 to 3 additional heteroatoms each of which independently is N, S, or O;

each of the Rc groups independently is halogen, alkyl, haloalkyl, hydroxyl, hydroxyalkyl, alkoxy, carbonyl, carboxyl, amino, nitrile, sulfonyl, sulfmyl, sulfhydryl, aryl, cycloalkyl, heteroaryl, or heterocyclyl;

each optional R ₂ group can be the same as or different from the other and independently is halogen, cyano, amino, alkyl, alkenyl, alkynyl, or cylcoalkyl;

R ₃ is hydrogen, alkyl, alkenyl, alkynyl, cylcoalkyl, or aryl; or

R ₃ and an R ₂ group on the ring atom adjacent to the ring atom to which -C(0)NR ₃ R4 is attached together form a C _1-5 alkylene or alkenylene linker;

R ₅ is hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, hydroxylalkyl, alkoxy, carboxyl, or carbonyl;

R6 is hydrogen, aryl, aminocarbonyl, alkyl, alkoxy, carbonyl, cycloalkyl, heteroaryl, or heterocyclyl, and when applicable 5 is optionally substituted with 1 to 4 Rd groups; each Rd is independently halogen, alkyl, haloalkyl, hydroxyl, hydroxyalkyl, alkoxy, carbonyl, carboxyl, amino, nitrile, sulfonyl, sulfmyl, sulfhydryl, aryl, cycloalkyl, heteroaryl, or heterocyclyl; or two Rd groups, together with the same atom to which they are attached, form a 4- to 8-membered heterocyclyl or heteroaryl; or

R ₃ and R4, together with the nitrogen atom to which they are attached, form an optionally substituted 3- to 7-membered heterocyclic ring with additional 0 to 3 heteroatoms each of which independently is N, S, or O.

[08] In some each of X and

2

X independently is N or CRe; and Re is hydrogen or lower alkyl. [09] In some other embodiments, A is and each of

X ¹ and X ² independently is N, CH, or C(CH ₃ ). Examples of these embodiments include those in which X ¹ is N, CH, or C(CH ₃ ); and X ² is N or CH.

e other embodiments, A is or and A is optionally substituted with one or more Ri groups on the ring not bonded to B.

[011] Specific examples of A include

[012] In some embodiments, each Ri independently is halogen, cyano, optionally substituted amino, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cyclyl, optionally substituted heterocyclyl, or NRaRb, and when applicable Ri is optionally substituted with one or two Rc groups.

[013] In some other embodiments, each Ri independently is halogen, cyano, optionally substituted amino, optionally substituted lower alkyl, optionally substituted phenyl, optionally substituted thienyl, optionally substituted pyridinyl, optionally substituted pyrrazolyl, optionally substituted piperidinyl, optionally substituted morpholinyl, optionally substituted piperazinyl, or optionally substituted pyrrolidinyl; and each optional substituent on Ri independently is halogen, lower alkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, or phenyl. Specific examples of such Ri include F, CI, Br, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, phenyl, morphalinyl, piperazinyl, and pyridine; when applicable Ri is optionally substituted with one or two Rc groups, and each Rc independently is halogen, lower alkyl, or lower haloalkyl.

[014] In some other embodiments, Ri is a 5-membered optionally substituted heterocyclyl or heteroaryl, and Ri has 1 to 3 heteroatoms each independently being O, S, or N. Examples of

such Ri include

; and Ri is optionally substituted with halogen, cyano, amino, lower alkyl, lower haloalkyl, or lower hydroxyalkyl.

[015] In still some other embodiments, Ri n is 0, 1 , 2, or 3; X° is O, CH ₂ , or NH; and Ri is optionally substituted with halogen, cyano, amino, lower alkyl, lower haloalkyl, or lower hydroxyalkyl.

[016] In some embodiments, Ra is H and Rb is optionally substituted aryl.

[017] In some other embodiments, each Rc independently is halogen, haloalkyl, alkyl, hydroxyalkyl, alkoxy, or heterocyclyl.

[018] In some embodiments, B is ^¾ —NH, _-NH, ^N --NH, —NH, -O , ^N — O , ; at least a heteroatom in B is not bonded to A or

-C(0)NR ₃ R4; and B is optionally substituted with one or more R ₂ groups.

and B is optionally substituted with one or more R ₂ groups.

[020] In a narrower set of embodiments, B is ; and B is bonded to -C(0)NRaRb on the ring atom adjacent to the nitrogen ring atom. In these embodiments, R ₂ can be halogen, cyano, amino, alkyl, alkenyl, alkynyl, or cylcoalkyl, or R ₂ and R ₃ together form a Ci-5 alkylene or alkenylene linker.

[021] In some embodiments, R ₃ is hydrogen or lower alkyl; or R ₃ and an R ₂ group on the ring atom adjacent to the ring atom to which -C(0)NR ₃ R4 is bonded, together form a C _1-5 alkylene or alkenylene linker.

[022] In a narrower set of embodiments, R ₃ is hydrogen; or R ₃ and an R ₂ group on the ring atom adjacent to the ring atom to which -C(0)NR ₃ R4 is attached, together form a C _2-3 alkylene linker.

[023] In some embodiments, R4 is alkyl and is optionally substituted with hydroxyl, aminocarbonyl, or aryl further optionally substituted with halogen, alkyl, or alkoxy.

[024] In alternative embodiments, R ₃ and R ₄ , together with the nitrogen atom to which they are attached, form an optionally substituted 3- to 7-membered heterocyclic ring with additional 0 to 3 heteroatoms each of which independently is N, S, or O. For example, R ₃ and R4, together with the nitrogen atom to which they are attached, can form pyrrolidinyl optionally substituted with alkyl, halogen, cyano, amino, aryl, heteroaryl, cyclyl, or heterocyclyl.

[025] In some embodiments, the compounds of this invention are of Formula II, Formula ΙΓ, Formula III, or Formula ΙΙΓ as shown below:

II ΙΓ III

[026] In a narrower set of embodiments, the compounds of this invention are of Formula Il-a or Formula Il-b as shown below:

II-a Il-b

In Formula II-a, m is 0, 1 , 2, or 3.

[027] In some embodiments of this invention, R ₂ is H or CH ₃ , and R ₃ is H, CH ₃ , or alkenyl.

[028] In some other embodiments, R ₂ and R ₃ together form a linker of CH ₂ , -CH ₂ CH ₂ -, or — CH ₂ CH ₂ CH ₂ — .

[029] In some embodiments, R4 is ^R5 . In some instances, R5 is hydrogen, alkyl, or hydroxylalkyl (e.g., -CH ₂ OH or -CH ₃ ). In some other instances, R6 is aryl or aminocarbonyl and is optionally substituted with one or two Rd groups.

[030] In some embodiments, X ¹ is N, CH, or C(CH ₃ ); each Ri, when present, independently is

halog or NRaRb; and when applicable Ri is optionally substituted with one or two Rc groups;

n is 0, 1 , 2, or 3;

X ⁶ is O, CH ₂ , or NH;

Ra is H and Rb is aryl;

each Rc independently is halogen, CF ₃ , alkyl, alkoxy, or hydroxyalkyl;

R ₂ is H or CH ₃ ;

R ₃ is H, CH ₃ , or alkenyl; or R ₂ and R ₃ together form a linker of-CH ₂ CH ₂ - or

CH ₂ CH ₂ CH ₂ ; R5 is alkyl or hydroxyalkyl;

R<5 is aryl optionally substituted with one or two Rd groups; and each Rd is independently halogen, alkyl, haloalkyl, alkoxy, aryl, or hydroxyalkyl.

[031] In some other embodiments, R ₅ is H and R6 is aryl optionally substituted with one or two Rd groups.

[032] In still some other embodiments, the compounds of Formula I have one of the following formulae:

XXX

In these formulae, X is CH or N; Ri, R ₅ and 5 are as described above (e.g., R ₅ is -CH ₂ OH and

[033] Specific examples of the compound include

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-phen yl-lH-pyrazolo[3,4- b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-( 1 -(3 -chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(3 -chlorophenyl)- 1 H- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-( 1 - (3 -chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(3 -fluorophenyl)- 1 H- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-l -methyl-4-(5-phenyl-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide; (S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(2-c hlorophenyl)-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(2-f luorophenyl)-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-N-methyl- 4-(5-phenyl-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-4-(5-bromo-lH-pyrazolo[3,4-b]pyridin-3-yl)-N-(l-(3-chlor o-4-fluorophenyl)-2- hydroxy ethyl)- 1 H-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(thi ophen-2-yl)-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(3- (trifluoromethyl)phenyl)- 1 H-pyrazolo[3 ,4-b]pyridin-3 -yl)- 1 H-pyrrole-2-carboxamide;

(R)-4-(5 -phenyl- 1 H-pyrazolo [3 ,4-b]pyridin-3 -yl)-N-( 1 -phenylethyl)- 1 H-pyrrole-2- carboxamide;

(S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(5-phenyl-lH-pyr azolo[3,4-b]pyridin- 3 -yl)- 1 H-pyrrole-2-carboxamide;

(S)-N-(2-hydroxy- 1 -phenyl ethyl)-4-(5-phenyl- 1 H-pyrazolo[3 ,4-b]pyridin-3-yl)- 1 H- pyrrole-2-carboxamide;

(4-(5-phenyl- 1 H-pyrazolo[3 ,4-b]pyridin-3-yl)- 1 H-pyrrol-2-yl)(pyrrolidin- 1 - yl)methanone;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(4-c hlorophenyl)-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(pyr idin-3-yl)-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-4-(5-(3-chloro-4-fluorophenyl)-lH-pyrazolo[3,4-b]pyridin -3-yl)-N-(l-(3-chloro- 4-fluorophenyl)-2-hydroxyethyl)- 1 H-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(4-f luorophenyl)-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-4-(5-(4-chloro-2-fluorophenyl)-lH-pyrazolo[3,4-b]pyridin -3-yl)-N-(l-(3-chloro- 4-fluorophenyl)-2-hydroxyethyl)- 1 H-pyrrole-2-carboxamide; (R)-N-(l-(3-chlorophenyl)ethyl)-4-(5-phenyl-lH-pyrazolo[3,4- b]pyridin-3-yl)-lH- pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-isop ropyl-lH-pyrazolo[3,4- b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(5-c hloropyridin-3-yl)-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(R)-N-( 1 -(3-methoxyphenyl)ethyl)-4-(5 -phenyl- 1 H-pyrazolo[3 ,4-b]pyridin-3 -yl)- 1 H- pyrrole-2-carboxamide;

(R)-4-(5-phenyl-lH-pyrazolo[3,4-b]pyridin-3-yl)-N-(l-phenylp ropyl)-lH-pyrrole-2- carboxamide;

(R)-N-(l-(4-chlorophenyl)ethyl)-4-(5-phenyl-lH-pyrazolo[3,4- b]pyridin-3-yl)-lH- pyrrole-2-carboxamide;

(S)-4-(5-(3-chlorophenyl)-lH-pyrazolo[3,4-b]pyridin-3-yl)-N- (l-(3-chlorophenyl)-2- hydroxy ethyl)- 1 H-pyrrole-2-carboxamide;

(S)-4-(5-(2-chloro-4-fluorophenyl)-lH-pyrazolo[3,4-b]pyridin -3-yl)-N-(l-(3- chlorophenyl)-2-hydroxyethyl)- 1 H-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(o-t olyl)-lH-pyrazolo[3,4- b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(l -methyl- lH-pyrazol-4- yl)- 1 H-pyrazolo[3 ,4-b]pyridin-3-yl)- 1 H-pyrrole-2-carboxamide;

(S)-4-(5-( 1 H-pyrazol-4-yl)- 1 H-pyrazolo[3 ,4-b]pyridin-3-yl)-N-( 1 -(3-chloro-4- fluorophenyl)-2-hydroxyethyl)- 1 H-pyrrole-2-carboxamide;

(4-(5-phenyl- 1 H-pyrazolo[3 ,4-b]pyridin-3-yl)- 1 H-pyrrol-2-yl)(2-phenylpyrrolidin- 1 - yl)methanone;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(m-t olyl)-lH-pyrazolo[3,4- b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-morp holino-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(phe nylamino)-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide; (S)-N-( 1 -(3 -chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(piperidin- 1 -yl)- 1 H- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(pip erazin-l-yl)-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-( 1 -(3 -chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(pyrrolidin- 1 -yl)- 1 H- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(5-morpholino-lH -pyrazolo[3,4- b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-( 1 -(3 -chlorophenyl)-2-hydroxyethyl)-4-(5-( 1 -methyl- 1 H-pyrazol-4-yl)- 1 H- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(5-isopropyl-lH- pyrazolo[3,4- b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(lH-pyr azolo[3,4-b]pyridin-3- yl)- 1 H-pyrrole-2-carboxamide;

(S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(5-phenyl-lH-pyr rolo[2,3-b]pyridin-3- yl)- 1 H-pyrrole-2-carboxamide;

(S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-phen yl-lH-pyrrolo[2,3- b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(R)-N-( 1 -(3-chlorophenyl)ethyl)-4-(5-phenyl- 1 H-pyrrolo[2,3-b]pyridin-3-yl)- 1 H- pyrrole-2-carboxamide;

(S)-N-( 1 -(3 -chlorophenyl)-2-hydroxyethyl)-4-(5 -( 1 -methyl- 1 H-pyrazol-4-yl)- 1 H- pyrrolo[2,3-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(R)-N-( 1 -(3-chlorophenyl)ethyl)-4-(5-( 1 -methyl- 1 H-pyrazol-4-yl)- 1 H-pyrrolo[2,3 - b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(R)-N-(l-(3-chlorophenyl)ethyl)-4-(5-(l-(2-hydroxyethyl)-lH- pyrazol-4-yl)-lH- pyrrolo[2,3-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(R)-4-(5-(lH-pyrazol-4-yl)-lH-pyrrolo[2,3-b]pyridin-3-yl)-N- (l-(3- chlorophenyl)ethyl)- 1 H-pyrrole-2-carboxamide;

(S)-4-(5-( 1 H-pyrazol-4-yl)- 1 H-pyrrolo[2,3 -b]pyridin-3-yl)-N-( 1 -(3-chlorophenyl)-2- hydroxy ethyl)- 1 H-pyrrole-2-carboxamide; (S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(5-(pyrrolidin-l -yl)-lH-pyrrolo[2,3- b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(5-morpholino-lH -pyrrolo[2,3- b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(2-methyl-5-phen yl-lH-pyrrolo[2,3- b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l -(3 -chlorophenyl)-2-hydroxyethyl)-4-(2-methyl-5-(l -methyl- lH-pyrazol-4- yl)- 1 H-pyrrolo[2,3-b]pyridin-3-yl)- 1 H-pyrrole-2-carboxamide;

(S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(5-isopropyl-2-m ethyl-lH-pyrrolo[2,3- b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(2-methyl-5-(2-m ethylpyridin-4-yl)- lH-pyrrolo[2,3-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(2-methyl-5-morp holino-lH- pyrrolo[2,3-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide;

(S)-2-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-7-(5-phen yl-lH-pyrazolo[3,4- b]pyridin-3-yl)-3,4-dihydropyrrolo[ 1 ,2-a]pyrazin- 1 (2H)-one;

(R)-2-(l-(3-chlorophenyl)ethyl)-7-(5-phenyl-lH-pyrrolo[2,3-b ]pyridin-3-yl)-3,4- dihydropyrrolo[ 1 ,2-a]pyrazin- 1 (2H)-one;

(R)-2-(l-(3-chlorophenyl)ethyl)-7-(5-phenyl-lH-pyrazolo[3,4- b]pyridin-3-yl)-3,4- dihydropyrrolo[ 1 ,2-a]pyrazin- 1 (2H)-one;

(S)-2-(l-(3-chlorophenyl)-2-hydroxyethyl)-7-(5-phenyl-lH-pyr rolo[2,3-b]pyridin-3- yl)-3 ,4-dihydropyrrolo[ 1 ,2-a]pyrazin- 1 (2H)-one;

(S)-N-( 1 -(3 -chlorophenyl)-2-hydroxyethyl)-4-(3 -(2-methylpyridin-4-yl)- 1 H- pyrazolo[3,4-b]pyridin-5-yl)-112-pyrrole-2-carboxamide;

(S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(3-(pyridin-4-yl )-lH-indazol-5-yl)-112- pyrrole-2-carboxamide;

(S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(3-(pyridin-4-yl )-lH-pyrazolo[3,4- b]pyridin-5-yl)-lH-pyrrole-2-carboxamide; and

N-(2-oxo-2-(4-(4-(pyrimidin-2-yl)phenyl)piperazin-l-yl)ethyl )-4-(3-(pyridin-4-yl)- lH-indazol-5-yl)-lH-pyrrole-2-carboxamide. [034] In another aspect, the present invention provides pharmaceutical compositions each comprising a compound of this invention as described above (e.g., a compound of Formula I disclosed herein) and a pharmaceutically acceptable carrier. In some embodiments, each of the compositions further includes an additional therapeutic agent. Examples of such therapeutic agent include, but are not limited to, a chemotherapeutic or anti-proliferative agent, an antiinflammatory agent, an immunomodulatory or immunosuppressive agent, an agent for treating a neurological disorder, an agent for treating cardiovascular disease, an agent for treating destructive bone disorders, an agent for treating liver disease, an anti-viral agent, an agent for treating blood disorders, an agent for treating diabetes, and an agent for treating

immunodeficiency disorders.

[035] Still in another aspect, the present invention relates to a method of treating a disease, disorder, or condition mediated by ERK or associated with ERK activity in a patient, which comprises administering to the patient in need thereof a therapeutically effective amount of a compound or pharmaceutical composition of this invention. Yet still another aspect of this invention provides using a compound of this invention for the manufacture of a medicament for the treatment of the disease, disorder, or condition mediated by ERK or associated with ERK activity.

[036] Such a disease, disorder, or condition is usually alleviated by a decrease of ERK activity. Examples of such a disease, disorder, or condition include but are not limited to cancers, immune dysfunction, hyper-inflammatory disorder, hyper-proliferative disorder, cardiac disorder, neurodegenerative disorder, a condition associated with organ transplant, immunologically mediated disorder, bone disorder, pancreatitis, kidney disease, pain, diseases related to vasculogenesis or angiogenesis, acute and chronic inflammatory disease, skin disease, diabetes, diabetic retinopathy, retinopathy of prematurity, age-related macular degeneration, asthma, septic shock, T-cell mediated diseases, and chronic obstructive pulmonary disease. In particular, the disease can be a cancer, which may be originated from brain, lung, colon, breast, gastric, pancreatic, head and neck, esophageal, renal, kidney, ovarian, skin, prostate, testicular, gynecological or thyroid.

[037] The invention also provides kits comprising a compound disclosed herein or a pharmaceutically acceptable salt, solvate, or prodrug thereof, packaging, and instructions for use thereof. Such kits can be used for the treatment or prevention in an individual of a disease or condition mediated by ERK. In some embodiments, the kit comprises a pharmaceutical formulation which includes a compound of this invention (e.g., a compound of Formula I) and packaging.

[038] Set forth below is a detailed description of the compounds of this invention, methods and processes for making, testing, and using these compounds which also constitute part of this invention.

Detailed Description of the Invention

[039] As use herein, unless clearly indicated otherwise, use of the terms "a", "an" and the like refers to one or more.

[040] Reference to "about" a value or parameter herein includes (and describes) embodiments that are directed to that value or parameter per se. For example, description referring to "about X" includes description of "X."

[041] As used herein, the word "or" has the meaning of both "or" and "and" and is equivalent to "and/or" - unless otherwise specifically limited to just "or."

[042] As used herein, the term "halo" or "halogen,"' by itself or as part of another substituent (e.g„ haloalkyl), refers to and includes fluoro, chloro, bromo, or iodo.

[043] As used herein, the term "alkyl," by itself or as part of another substituent (which usually takes the short form of "alk,"e.g., alkoxy), refers to and includes saturated linear (i.e.

unbranched) or branched hydrocarbon radicals, having the number of carbon atoms designated (e.g., Ci-io means one to ten carbons). Particular alkyl groups include those having 1 to 10 carbon atoms (a "Ci_io alkyl"). More particular alkyl groups are those having 1 to 6 carbon atoms (a "Ci_6 alkyl"), 1 to 4 carbon atoms (a "Ci_4 alkyl") , 1 to 3 carbon atoms (a "Ci_3 alkyl") or 1 to 2 carbon atoms (a "Ci_2 alkyl"). Examples of "Ci_io alkyl" include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, n- heptyl, n-octyl, n-nonyl, n-decyl and the like. An alkyl group can be optionally substituted with such substituents as halogen, cyano, amino, hydroxyl etc. As used herein, the term "lower alkyl" refers to alkyl of 1 to 6 carbon atoms which are optionally substituted with one or more appropriate substituents such as halogen, amino, cyano, or hydroxyl.

[044] As used herein, the term "alkenyl," by itself or as part of another substituent, refers to and includes unsaturated linear (i.e. unbranched) or branched hydrocarbon radicals containing at least one carbon-carbon double bond, having the number of carbon atoms designated (e.g., C _2-10 means two to ten carbons). Particular alkenyl groups are those having 2 to 10 carbon atoms (i.e., a "C _2-10 alkenyl"). More particular alkenyl groups are those having 2 to 8 carbon atoms (a "C2-8 alkenyl") or 2 to 6 carbon atoms (a "C2-6 alkenyl"). Examples of "C _2-10 alkenyl" include, but are not limited to, ethenyl, 1 -propenyl, 2-propenyl, 1 -methylethenyl, 2-methyl- 1 -propenyl, 2-methyl- 2-propenyl, 2-methyl-3-butenyl, 3-methyl-3-butenyl, 1 ,2-dimethyl- 1 -propenyl, and 1 ,2-dimethyl- 2-propenyl. As used herein, the term "lower alkenyl" refers to alkenyl of 1 to 6 carbon atoms which are optionally substituted with one or more appropriate substituents such as halogen, amino, cyano, or hydroxyl.

[045] As used herein, the term "aminocarbonyl" refers to NRR'-C(=0)- wherein each of R and R' independently can be hydrogen, lower (e.g., C _1-6 ) alkyl or alkenyl which may be optionally substituted with halogen or cyano.

[046] As used herein, the term "heteroatom" refers to "S", "O" or "N" in a ring which can be saturated, unsaturated, or aromatic. The "N" heteroatom can be optionally subtitled with alkyl or alkenyl.

[047] As used herein, the term "cycloalkyl" or "cyclyl", by itself or as part of another substituent, refers to and includes saturated monocyclic hydrocarbon radicals, having the number of carbon atoms designated (e.g., C _3-10 means three to ten carbons). Particular examples of cycloalkyl or cyclyl groups include those having 3 to 10 carbon atoms (a "C _3-10 cycloalkyl"). More particular cycloalkyl groups include those having 3 to 8 carbon atoms (a "C3_8 cycloalkyl"), 3 to 6 carbon atoms (a "C _3-6 cycloalkyl") or 4 to 5 carbon atoms (a "C4.5 cycloalkyl"). Examples of "C _3- io cycloalkyl" include, but are not limited to, cyclopropyl, cyclopentyl, cyclohexyl, and the like.

[048] As used herein, the term "aryl" or "aryl group", by itself or as part of another substituent, refers to and includes monocyclic or polycyclic aromatic hydrocarbon radicals, having the number of annular carbon atoms designated (e.g., C _6-14 means six to fourteen carbons).

Particular aryl groups are those having 6 to 14 annular carbon atoms (a "C6 _-14 aryl"). Examples of "C6-14 aryl" include, but are not limited to, phenyl, naphthyl, anthracenyl, and the like. In some embodiments, an aryl may contain a single ring (e.g., phenyl). In some embodiments, an aryl may contain multiple (e.g., two or three) rings. In some embodiments, an aryl may contain multiple condensed rings where at least one of the condensed rings is aromatic (e.g., 1,2,3,4- tetrahydronaphthyl and naphthyl).

[049] As used herein, a combined term such as "arylalkyl" denotes a group including aryl and alkyl wherein aryl is a substituent on alkyl.

[050] As used herein, the term "heterocyclyl" or "heterocycle" refers to monocyclic or bicyclic radicals which may be fully saturated, partially saturated, or fully unsaturated or aromatic, having the number of annular carbon atoms designated (e.g., C _3-10 means three to ten annular carbon atoms) and containing at least one or more of the same or different heteroatoms selected from N, S or O, provided that at least one annular carbon atom is present and two annular oxygen atoms, if present, do not occupy directly neighboring positions. A "heterocyclyl" or

"heterocycle" may be a 3 to 15-membered saturated or partially unsaturated ring containing 1 to 4 heteroatoms selected from O, S and N, where the ring may be monocyclic, bicyclic or tricyclic, contain at least one annular carbon atom and 1 to 3 nitrogen atoms, and/or 1 oxygen or sulfur atom or 1 or 2 oxygen and/or sulfur atoms; provided that when more than one annular oxygen atoms are present, they do not occupy directly neighboring positions. Examples of

"heterocyclyl" or "heterocycle" include, but are not limited to, 2-oxiranyl, 2-aziridinyl, 2- tetrahydrofuranyl, 3-tetrahydrofuranyl, 2-tetrahydrothienyl, 3-tetrahydrothienyl, 2-pyrrolidinyl, 3-pyrrolidinyl, 3-isoxazolinyl, 4-isoxazolinyl, 5-isoxazolinyl, 3-isothiazolinyl, 4-isothiazolinyl, 5-isothiazolinyl, 3-pyrazolinyl, 4-pyrazolinyl, 5-pyrazolinyl, 2-oxazolinyl, 4-oxazolinyl, 5- oxazolinyl, 2-thiazolinyl, 4-thiazolinyl, 5-thiazolinyl, 2-imidazolinyl, 4-imidazolinyl, 1 ,2,4- oxadiazol-3-yl, l,2,4-oxadiazol-5-yl, l,2,4-thiadiazol-3-yl, l,2,4-thiadiazol-5-yl, l,2,4-triazol-3-yl, l,3,4-thiadiazol-2-yl, l,3,4-oxadiazol-2-yl, l,3,4-triazol-2-yl, 2,3-dihydrofuran-2-yl, 2,3- dihydrofuran-3-yl, 2,4-dihydrofuran-2-yl, 2,4-dihydrofuran-3-yl, 2,3-dihydrothiophen-2-yl, 2,3- dihydrothiophen-3-yl, 2,4-dihydrothiophen-2-yl, 2,4-dihydrothiophen-3-yl, 2-pyrrolin-2-yl.

[051] As used herein, the term "heteroaryl" refers to aromatic heterocyclyl or heterocycle as defined herein. Examples of "heteroaryl" include, but are not limited to, 2-furanyl, 3-furanyl, thiophen-2-yl, thiophen-3-yl, lH-pyrrol-2-yl, lH-pyrrol-3-yl, isoxazol-3-yl, isoxazol-4-yl, isoxazol-5-yl, isothiazol-3-yl, isothiazol-4-yl, isothiazol-5-yl.

[052] As used herein, the term "hydroxyalkyl" refers to alkyl group with at least one hydroxyl substituents. [053] As used herein, the term "substituted" whether preceded by the term "optionally" or not, refers to replacement of hydrogen radicals in a given structure with the radical of a specified substituent. Specific substituents are described above in the definitions and below in the description of compounds and examples thereof. Unless otherwise indicated, an optionally substituted group can have a substituent at each substitutable position of the group, and when more than one position in any even structure can be substituted with more than one substituent selected from a specified group, the substituent can be either the same or different in every position. A ring substituent, such as heterocycloalkyl, can be bound to another ring, such as a cycloalkyl to form a spiro-bicyclic ring system, e.g., both rings share one common atom. As one or ordinary skill in the rat will recognize, combinations of substituents envisioned by this disclosure are those combinations that result in the formation of stable or chemically feasible compounds. For convenience and as commonly understood, the term "optionally substituted" is used interchangeably with the phrase "substituted or unsubstituted" and only applies to the chemical entities that can be substituted with. As describe herein, when the term "optionally substituted" precedes a list, this term refers to all of the subsequent substitutable group sin that list.

[054] As used herein, the term "therapeutically effective amount" means that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue, system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other clinician.

[055] As used herein, the term "treatment" or "treating" refers to the treatment of a mammal afflicted with a pathological condition and refers to an effect that alleviates the condition, e.g., by killing the cancerous cells, but also to an effect that results in the inhibition of the progress of the condition, and includes a reduction in the rate of progress, a halt in the rate of progress, amelioration of the condition, and cure of the condition.

[056] As used herein, the term "pharmaceutically acceptable" pertains to 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 a subject (e.g. human) without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. Each carrier, excipient, etc. must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation. [057] As used herein, the term "pharmaceutically acceptable salt" - unless otherwise specified - refers to salts which are suitable for use in contact with the tissues of a subject (e.g., human) without excessive adverse effect. In some embodiments, pharmaceutically acceptable salts include salts of a compound of the invention having an acidic group (e.g., potassium salts, sodium salts, magnesium salts, calcium salts) or a basic group (e.g., sulfate, hydrochloride, phosphate, nitrate, carbonate).

[058] As used herein, the term "patient" refers to a mammal which include humans and non- human mammals such as cows.

[059] Unless specifically otherwise defined, all the terms used herein have their common meanings as known to a skilled person in the art.

Synthesis of Compounds

[060] Set forth below are some exemplary schemes of methods that have been used or can be used for synthesizing the compounds of this invention:

Scheme A:

[061] In Scheme A, compound I reacts with iodine in an organic solvent (such as dioxane) in the presence of a base (such as sodium hydroxide) to give rise to compound II which then reacts with SEM-Cl in the presence of a base (such as t-butoxy potassium) to provide compound III. Compounds III and IV react in the presence of a catalyst to produce compound V which in turn is converted to compound VI in a catalytic reaction. Compound VI is hydrolyzed in the presence of a base such as sodium hydroxide to give an acid of formula VII. The product is coupled with compound VIII in the presence of EDCL and alcohol to give rise to compound IX which finally is reduced, e.g., in the presence of TFA, to give compound X of this invention.

Scheme B:

[062] In Scheme B, compound XI reacts with iodine in an organic solvent (e.g., DMF) in the presence of a base (e.g., potassium hydroxide) to give compound XII which then reacts with SEM-C1 in the presence of a strong base (e.g., t-butoxy potassium) to produce compound XIII. Compounds XIII and XIV then react in the presence of a catalyst to give a coupled compound XV which then go through a replacement reaction to give compound XVI. This product is then hydrolyzed in a base condition to give rise to a carboxylic acid XVII which then undergoes a couple reaction to give compound XIX. Compound XIX is then reduced to give a compound XX of this invention.

Scheme C:

[063] In Scheme C, compound XXI is first iodinized and then reacts with SEM-C1 in the presence of a strong base (e.g., potassium t-butoxy). The compound is then coupled with compound XXIV, in the presence of a catalyst, to provide compound XXV, which then undergoes a replacement reaction to provide compound XXVI having. Compound XXVI is then hydrolyzed under basic conditions (e.g., in the presence of sodium hydroxide) to give a carboxylic acid compound XXVII which then reacts with an amino compound to give an amide. The amide is finally reduced to give the compound XXX of this invention.

[064] The present invention is further exemplified by the following examples that illustrate the preparation of the compounds of the invention. These examples are for illustration only, and do not intend to limit to scope of the present invention in any way.

Example 1: Synthesis of methyl 4-(5-(phenylamino)-l-((2-(trimethylsilyl)ethoxy)methyl)- lH-pyrazolo[3,4-blpyridin-3-yl)-l-tosyl-lH-pyrrole-2-carboxy late

[065] The title compound was synthesized according to Scheme A with the following detailed. Step 1. Preparation of 5-bromo-3-iodo- -pyrazolo[3,4-b] pyridine (II) [066] To a solution of 5-bromo-lH-pyrazolo[3,4-b]pyridine (I) (5 g, 25.25 mmol, 1 eq) in 1,4- dioxane (100 mL) and 4 N aqueous NaOH (100 mL) was added iodine (64.1 g, 252.5 mmol, 10 eq). The mixture was stirred at 60 °C overnight, and TLC showed the reaction was complete. The reaction mixture was extracted with EtOAc (100 ml x 2). The combined organic extracts were washed with saturated aqueous NaHS0 ₃ solution (100 mL x 3) and brine (50 mL), dried over Na ₂ S0 ₄ , filtered, and concentrated to give the title compound 5-bromo-3-iodo-lH- pyrazolo[3,4-b]pyridine as an off-white solid (7.5 g , yield:91%).

Step 2. Preparation of 5-bromo-3-iodo-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrazolo[3,4-b]pyridine (III)

M

[067] 5-bromo-3-iodo-lH-pyrazolo[3,4-b]pyridine prepared in Step 1 was used without any further purification. To a solution of 5-bromo-3-iodo-lH-pyrazolo[3,4-b]pyridine (7.5 g, 23.15 mmol, 1 eq) and t-BuOK (3.12 g, 27.78 mmol, 1.2 eq) in DMF (75 mL) was added SEMC1 (4.63 g, 27.78 mmol, 1.2 eq) dropwise at 0 °C. The mixture was allowed to warm to room temperature and stirred overnight. TLC showed the reaction was complete. The reaction mixture was extracted with EtOAc (100 ml x 2). The combined organic extracts were washed with Na ₂ C0 ₃ (100 ml x 2) and brine (50 ml), dried over anhydrous Na ₂ S0 ₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (petroleum ether : EtOAc = 20: 1) to give the title compound 5-bromo-3-iodo-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyraz olo[3,4- bjpyridine as an off-white solid (7.5 g, yield:71.3%).

Step 3. Preparation of Methyl 4-(5-bromo-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrazolo [3,4-b]pyridin-3-yl) -l-tosyl-lH-pyrrole-2-carboxylate (V)

[068] A solution of 5-bromo-3-iodo-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyraz olo[3,4-b] pyridine from Step 2 (5 g, 11 mmol, 1 eq), methyl 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)-l-tosyl-lH-pyrrole-2-carboxylate (IV) (5.27 g, 13 mmol, 1.2 eq), Pd(dppf)Cl ₂ (402 mg, 0.55 mmol, 0.05 eq.), Na ₂ C0 ₃ (2.322 g, 22 mmol, 2 eq.) in DMF (50 mL) and H ₂ 0 (5 mL) was stirred at 50 °C for 140 minutes. The reaction mixture was filtered, and the filtrate was extracted with EtOAc (100 mL x 3). The combined organic extracts were washed with H ₂ 0 (100 ml x 3) and brine (50 ml x 2), dried over anhydrous Na ₂ S0 ₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (Petroleum ether : EtOAc = 20: 1-15: 1) to give the title compound methyl 4-(5-bromo-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrazolo[ 3,4- b]pyridin-3-yl) -l-tosyl-lH-pyrrole-2-carboxylate as an off-white solid (3.17 g, yield:47.6%). Step 4. Preparation of Methyl 4-(5-(pyridin-3-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrazolo[3,4-b]pyridin-3-yl)-l-tosyl-lH-pyrrole-2-carboxylat e

[069] A solution of methyl 4-(5-bromo-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrazolo[ 3,4- b]pyridin-3-yl)-l-tosyl-lH-pyrrole-2-carboxylate (290 mg, 0.48 mmol, 1.0 eq), pyridin-3- ylboronic acid (65 mg, 0.528 mmol, 1.1 eq.), Na ₂ C0 ₃ (102 mg, 0.960 mmol, 2.0 eq.), and Pd(dppf)Cl ₂ (35 mg, 0.048 mmol, 0.1 eq.) in DMF (6 mL) and H ₂ 0 (1 mL) was stirred under nitrogen at 65 °C for 100 minutes. The mixture was filtered. The filtrate was diluted with EA (50 mL) and water (20 mL). The organic layer was separated, washed with brine (20 mL x 3), dried over Na2S04, and concentrated. The residue was purified by silica gel column

chromatography (PE:EtOAc = 3: 1) to give the title compound methyl 4-(5-(pyridin-3-yl)-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrazolo[3,4-b]pyridin-3-y l)-l-tosyl-lH-pyrrole-2- carboxylate (134 mg, crude yield: 45%).

Step 5. Preparation of 4-(5-(pyridin-3-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxylic acid (VII)

[070] A solution of 4-(5-(pyridin-3-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrazolo[3,4- b]pyridin-3-yl)-l-tosyl-lH-pyrrole-2-carboxylate prepared from step 4 (134 mg, 0.222 mmol, 1.0 eq.) and NaOH (44 mg, 1.11 mmol, 5.0 eq.) in MeOH (1 mL) and H ₂ 0 (1 mL) was stirred at 90 °C for 4 hours. TLC showed the reaction was complete. The reaction mixture was cooled to the room temperature and concentrated to remove most MeOH. The resultant aqueous solution was acidified to pH about 3 with concentrated hydrochloric acid and extracted with EA (50 mL). The organic layer was washed with H ₂ 0 (20 mL) and brine (20 mL), dried over Na ₂ S0 ₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (MeOH:DCM = 10: 1) to give the title compound 4-(5-(pyridin-3-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxylic acid (67 mg, crude yield: 69%).

Step 6. Preparation of (S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(pyr idin-3- yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrazolo[3,4-b]p yridin-3-yl)-lH-pyrrole-2- carboxamide (IX)

[071] To a solution of 4-(5-(pyridin-3-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxylic acid prepared from step 5 (66 mg, 0.151 mmol), (5)-2-amino-2-(3-chloro-4-fluorophenyl)ethan-l-ol (34 mg, 0.181 mmol), EDCI (35 mg, 0.181 mmol), HOBt (24 mg, 0.181 mmol) in DCM (1.5 mL) was added DIPEA (58 mg, 0.453 mmol). The mixture was stirred at the room temperature overnight. The mixture was concentrated, and partitioned between EtOAc (50 mL) and H ₂ 0 (20 mL). The organic layer was collected, washed with brine (20 mL), dried over Na ₂ S0 ₄ , filtered, and concentrated to give the title compound (S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(pyr idin-3-yl)-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrazolo[3,4-b]pyridin-3-y l)-lH-pyrrole-2-carboxamide (76 mg, yield: 84%).

Step 7. Preparation of (S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(pyr idin-3- yl)-lH-pyrazolo[3,4-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide (X)

[072] A solution of (S)-N-(l-(3-chloro-4-fluorophenyl)-2-hydroxyethyl)-4-(5-(pyr idin-3-yl)-l- ((2-(trimethylsilyl)ethoxy)methyl)- 1 H-pyrazolo [3 ,4-b]pyridin-3 -yl)- 1 H-pyrrole-2-carboxamide prepared from step 6 (76 mg, 0.125 mmol) in DCM (1.5 mL) and TFA (1.5 mL) was stirred at the room temperature for 3 hours. TLC showed the reaction was complete. The mixture was concentrated to dryness and the residue was dissolved in MeOH (2 mL) and ammonia (28%, 2 mL), and stirred for 1.5 hours. The organic solvent was removed by concentration under vacuo and the resulting aqueous solution was extracted with EtOAc (50 mL). The EtOAc phase was collected, washed with brine (20 mL), dried by Na ₂ S0 ₄ , filtered, and concentrated. The residue was purified by column chromatography to give the title compound (S)-N-(l-(3-chloro-4- fluorophenyl)-2-hydroxyethyl)-4-(5-(pyridin-3-yl)-lH-pyrazol o[3,4-b]pyridin-3-yl)-lH-pyrrole- 2-carboxamide (4 mg yield: 7%).

Step 8. Preparation of methyl 4-(5-(phenylamino)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrazolo[3,4-b]pyridin-3-yl)-l-tosyl-lH-pyrrole-2-carboxylat e

[073] A solution of methyl 4-(5-bromo-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrazolo[ 3,4- b]pyridin-3-yl)-l-tosyl-lH-pyrrole-2-carboxylate (250 mg, 0.41 mmol), aniline (58 mg, 0.62 mmol), Tris(dibenzylideneacetone)dipalladium(0) - or Pd2(dba)3 - (37 mg, 0.041 mmol), Xantphos (2 mg. 0.041 mmol) and Cs ₂ C(¾ (267 mg, 0.82 mmol) in dioxane was stirred at 90 °C under nitrogen overnight. The mixture was diluted with EtOAc (50 mL), washed with water (30 mL x 2), dried over anhydrous sodium sulfate, and concentrated to give the title compound of this invention methyl 4-(5-(phenylamino)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-p yrazolo[3,4- b]pyridin-3-yl)-l-tosyl-lH-pyrrole-2-carboxylate (215 mg, yield: 84%). [074] Table 1 below lists exemplary compounds of this invention that were also synthesized largely according to Scheme A.

Table 1

Example 2: Synthesis of (R)-N-(l-(3-chlorophenyl)ethyl)-4-(5-(l-methyl-lH-pyrazol-4- yl)- lH-pyrrolo[2,3-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide

[075] The title compound was synthesized according To Scheme B as detailed below.

Step 1: Preparation of 5-bromo-3-iodo- -pyrrolo[2,3-b]pyridine

[076] To a solution of 5-bromo-lH-pyrrolo[2,3-b]pyridine (9.0 g, 0.046mol) and KOH (5.16 g, 0.092 mol) in DMF (90 mL) was added I ₂ (12.75 g, 0.050 mol) over an ice-water bath. The mixture was stirred at the room temperature for 4 hours, and TLC showed the reaction was complete. Saturated NaHS0 ₃ aqueous solution (50 mL) was added to quench the reaction. The precipitate that formed was collected by filtration, washed with water (50 mL), dried under vacuum to give the title compound 5-bromo-3-iodo-lH-pyrrolo[2,3-b]pyridine as a yellow solid (14.45 g, yield: 97%).

Step 2. Preparation of 5-bromo-3-iodo-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrrolo[2,3-b] pyridine (XIII)

M

[077] To a solution of 5-bromo-3-iodo-lH-pyrrolo[2,3-b]pyridine (XII) prepared by Step 1 (10 g, 0.031 mol) and KOtBu (6.22 g, 0.037 mol) in DMF (90 mL) was slowly added SEMCl (4.15 g, 0.037) over an ice-water bath slowly. The mixture was stirred at the room temperature overnight, and TLC showed the reaction was complete. After removal of DMF by concentration, the mixture was diluted with EtOAc (200 mL), washed by water (50 mL x 3), dried over Na ₂ S0 ₄ , filtered, and concentrated. The residue was purified by flash column chromatography

(Petroleum Ether : EtOAc = 20 : 1) to give the title compound 5-bromo-3-iodo-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridine as a yellow solid (10.77 g, yield:77%). Step 3. Preparation of methyl 4-(5-bromo-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrrolo[2,3-b]pyridin-3-yl) -l-tosyl-lH-pyrrole-2-carboxylate (XV)

[078] A suspension of 5-bromo-3-iodo-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrro lo[2,3- bjpyridine (XIII) (6.3 g, 13.9 mmol), methyl 4-(4,4,5,5-tetramethyl-l ,3,2-dioxaborolan-2-yl)-l- tosyl-lH-pyrrole-2-carboxylate (XIV) (5.35 g, 13.2 mmol), Pd(dppf)C12 (1.14 g, 1.39 mmol), and Na2C03 (2.95 g, 27.8 mmol) in DMF (65 mL) and water (6.5 mL) was stirred at 65-70 °C (internal temperature) under nitrogen atmosphere for 2 hours. The reaction mixture was poured into water (250 mL) with vigorous stirring. The precipitate was collected by filtration, washed with water (50 mL x 3). The filter cake was taken in EtOAc (250 mL), washed with water (50 mL x 3), dried over Na ₂ S0 ₄ , filtered, and concentrated. The residue was purified by column chromatography (petroleum ether:EtOAc = 10: 1) to give the title compound methyl 4-(5-bromo- 1 -((2-(trimethylsilyl)ethoxy)methyl)- 1 H-pyrrolo[2,3-b]pyridin-3-yl)- 1 -tosyl- lH-pyrrole-2- carboxylate (XV) as a white solid (5.0 g, yield: 44%).

Step 4: Preparation of methyl 4-(5-(l-methyl-lH-pyrazol-4-yl)-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridin-3-yl )-l-tosyl-lH-pyrrole-2- carboxylate (XVI)

[079] A mixture of methyl 4-(5-bromo-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2 ,3- b]pyridin-3-yl) -1 -tosyl- lH-pyrrole-2-carboxylate (XV) (4.4 g, 7.2 mmol), l-methyl-4-(4,4,5,5- tetramethyl-l ,3,2-dioxaborolan-2-yl)-lH-pyrazole (1.8 g, 8.64 mmol), Pd(dppf)Cl ₂ (588 mg, 0.72 mmol), and Na ₂ C0 ₃ (1.5 g, 14.4 mmol) in DMF (72 mL) and water (7.2 mL) was stirred at 75-80 °C (internal temperature) for 2 hours under nitrogen atmosphere. The reaction mixture was diluted with water (300 mL) and extracted with EtOAc (500 mL x 2). The combined organic layer was washed with brine (300 mL x 3), dried over Na ₂ S0 ₄ , filtered, and concentrated to give the crude title compound methyl 4-(5-( 1 -methyl- lH-pyrazol-4-yl)-l -((2-

(trimethylsilyl)ethoxy)methyl)- 1 H-pyrrolo[2,3-b]pyridin-3 -yl)- 1 -tosyl- 1 H-pyrrole-2-carboxylate (XVI), which was used in the next step without and further purification.

Step 5. Preparation of 4-(5-(l-methyl-lH-pyrazol-4-yl)-l-((2-

(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridin-3 -yl)-lH-pyrrole-2-carboxylic acid (XVII)

[080] To a solution of methyl 4-(5-(l -methyl- 1 H-pyrazol-4-yl)- 1 -((2-(trimethylsilyl)- ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridin-3-yl)-l -tosyl- lH-pyrrole-2-carboxylate (XVI) (crude, ~ 8.2 mmol) in THF (60 mL) was added a solution of NaOH (3.2 g, 82 mmol) in water (60 mL). The mixture was stirred under reflux for 18 hours. The mixture was concentrated to remove most of THF. The aqueous phase was adjusted to pH of 3-4 by concentrated hydrochloric acid. The resulting suspension was extracted with EtOAc (200 mL). The organic phase was washed with water (100 mL x 2), dried over Na ₂ S0 ₄ , filtered, and concentrated to give the crude title compound 4-(5-(l -methyl- lH-pyrazol-4-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrrolo[2,3-b]pyridin-3-yl)-lH-pyrrole-2-carboxylic acid (XVII) (3.0 g), which was used in the next step without any further purification.

Step 6. Preparation of (R)-N-(l-(3-chlorophenyl)ethyl)-4-(5-(l-methyl-lH-pyrazol-4- yl)-l- ((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridin- 3-yl)-lH-pyrrole-2- carboxamide (XIX)

[081] To a mixture of 4-(5-(l -methyl- lH-pyrazol-4-yl)-l-((2-(trimethylsilyl)ethoxy)-methyl)- lH-pyrrolo[2,3-b]pyridin-3-yl)-lH-pyrrole-2-carboxylic acid (XVII) (3.0 g, ~ 6.8 mmol), (R)-l- (3-chlorophenyl)ethanamine (XVIII) (1.28 g, 8.2 mmol), and HATU (3.12 g, 8.2 mmol) in DCM (30 mL) was added DIPEA (9 g, 70 mmol). The mixture was stirred at the room temperature overnight and concentrated. The residue was taken in EtOAc (50 mL), washed with water (30 mL), dried over Na ₂ S0 ₄ , and concentrated. The reside was purified by column chromatography (Petroleum ether : EtOAc = 4: 1) to give the title compound as a yellow solid (3.25 g, yield: 83%, Lot#: JSI111038-69-1).

Step 7. Preparation of (R)-N-(l-(3-chlorophenyl)ethyl)-4-(5-(l-methyl-lH-pyrazol-4- yl)- lH-pyrrolo[2,3-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide (XX)

[082] To a solution of (R)-N-(l-(3-chlorophenyl)ethyl)-4-(5-(l-methyl-lH-pyrazol-4- yl)-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridin-3-yl )-lH-pyrrole-2-carboxamide (XIX) (3.25 g, 5.7 mmol) in 1M TBAF/THF (35 mL) was added ethylenediamine (8.50 g, 140 mmol). The mixture was stirred under reflux for 48 hours and concentrated. The residue was taken in EtOAc (100 mL), washed with water (30 mL x 5), dried over Na ₂ S0 ₄ , filtered, and concentrated. The residue was purified by column chromatography (DCM : MeOH = 12: 1) to give the title compound (R)-N-(l-(3-chlorophenyl)ethyl)-4-(5-(l -methyl- lH-pyrazol-4-yl)- lH-pyrrolo[2,3- b]pyridin-3-yl)-lH-pyrrole-2-carboxamide (XX) as a yellow solid (1.38 g, yield: 54%).

[083] Table 2 below lists some exemplary compounds of this invention that were synthesized according to Scheme B. Table 2

Example 3: Synthesis of (S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(2-methyl-5-(2- methylpyridin-4-yl)-lH-pyrrolo[2,3-b]pyridin-3-yl)-lH-pyrrol e-2-carboxamide

[084] The title compound was synthesized according to Scheme C as follows:

Step 1. Preparation of 5-bromo-3-iod -2-methyl-lH-pyrrolo[2,3-b]pyridine (XXII)

[085] To a solution of 5-bromo-2-methyl-lH-pyrrolo[2,3-b]pyridine (XXI) (3.1 g, 14.7 mmol) in acetone (90 mL) was added NIS (3.6 g, 16.2 mmol). The mixture was stirred at the room temperature for 2 hours, and diluted with EtOAc (200 mL). The mixture was washed by H ₂ 0 (50 mL x 2) and brine (50 mL), dried over Na ₂ S0 ₄ , filtered, and concentrated to give the title compound 5-bromo-3-iodo-2-methyl-lH-pyrrolo[2,3-b]pyridine (XXII) as a yellow solid (4.4 g, yield: 89%).

Step 2. Preparation of 5-bromo-3-iodo-2-methyl-l-((2-(trimethylsilyl)ethoxy)methyl) -lH- pyrrolo[2,3-b] pyridine (XXIII)

[086] To a solution of 5-bromo-3-iodo-2-methyl-lH-pyrrolo[2,3-b]pyridine (XXII) (4.4 g, 13.1 mmol) in DMF (80 mL), cooled over an ice- water bath, was added t-BuOK (2.4 g, 21.0 mmol). After stirring for 0.5 hour at that temperature, SEMC1 (3.5 g, 21.0 mmol) added slowly. The mixture was stirred at the room temperature overnight, and diluted with H ₂ 0 (100 mL) and EtOAc (300 mL). The organic layer was washed with brine (50 mL x 3), dried over Na ₂ S0 ₄ , filtered, and concentrated. The residue was purified by column chromatography (Petroleum ether : EtOAc = 8: 1) to give the title compound 5-bromo-3-iodo-2-methyl-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridine (XXIII) as a yellow solid (3.6 g, yield: 60%).

[087] Step 3. Preparation of methyl 4-(5-bromo-2-methyl-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridin-3-yl )-l-tosyl-lH-pyrrole-2- carboxylate (XXV)

[088] A solution of 5-bromo-3-iodo-2-methyl-l-((2-(trimethylsilyl)ethoxy)methyl) -lH- pyrrolo[2,3-b]pyridine (XXIII) (2.33 g, 5.0 mmol), methyl 4-(4,4,5,5-tetramethyl-l ,3,2- dioxaborolan-2-yl)-l-tosyl-lH-pyrrole-2-carboxylate (XXIV) (2.02 g, 5.0 mmol), Pd(dppf)Cl ₂ (370 mg, 0.5 mmol), and a ₂ C0 ₃ (1.06 g, 10 mmol) in DMF/H ₂ 0 (60mL/12 mL) was stirred at 50 °C for 1 hour under nitrogen atmosphere. The mixture was diluted with ice-H ₂ 0 (300 mL) and the precipitate formed was collected by filtration and purified by column chromatography (Petroleum Ether : EtOAc =8 : 1) to give the title compound methyl 4-(5-bromo-2-methyl-l-((2- (trimethylsilyl)ethoxy)methyl)- 1 H-pyrrolo[2,3-b]pyridin-3 -yl)- 1 -tosyl- 1 H-pyrrole-2-carboxylate (XXV) as a yellow solid (1.2 g, 35% yield).

Step 4. Preparation of methyl 4-(2-methyl-5-(2-methylpyridin-4-yl)-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridin-3-yl )-l-tosyl-lH-pyrrole-2- carboxylate (XXVI)

[089] A solution of methyl 4-(5-bromo-2-methyl- 1 -((2-(trimethylsilyl)ethoxy)methyl)- 1 H- pyrrolo[2,3-b]pyridin-3-yl)-l-tosyl-lH-pyrrole-2-carboxylate (XXV) (200 mg, 0.324 mmol), 2- methylpyridin-4-ylboronic acid (53 mg, 0.388 mmol), Pd(dppf)Cl ₂ (23 mg, 0.032 mmol), and Na ₂ C0 ₃ (69 mg, 0.648 mmol) in DMF/H ₂ 0 (4 mL/0.4 mL) was stirred at 65 °C overnight under nitrogen atmosphere. The mixture was diluted with EtOAc (50 mL), washed with H ₂ 0 (15 mL x 2) and brine (30 mL), dried over Na ₂ S0 ₄ , and concentrated. The residue was purified by column Chromatography (Petrolem Ether : EtOAc = 4: 1 to 2: 1) to give the title compound methyl 4-(2- methyl-5-(2-methylpyridin-4-yl)- 1 -((2-(trimethylsilyl)ethoxy)methyl)- 1 H-pyrrolo[2,3 -bjpyridin- 3-yl)-l-tosyl-lH-pyrrole-2-carboxylate (XXVI) as a yellow solid ( 132 mg, yield: 65%).

Step 5. Preparation of 4-(2-methyl-5-(2-methylpyridin-4-yl)-l-((2-

(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridin-3 -yl)-lH-pyrrole-2-carboxylic acid (XXVII)

[090] To a solution of methyl 4-(2-methyl-5-(2-methylpyridin-4-yl)-l-((2- (trimethylsilyl)ethoxy)methyl)- 1 H-pyrrolo[2,3-b]pyridin-3 -yl)- 1 -tosyl- 1 H-pyrrole-2-carboxylate

(XXVI) (132 mg, 0.21 mmol) in MeOH (6 mL) was added NaOH (4 N, 2 mL). The mixture was stirred at 90 °C overnight and concentrated to remove MeOH. The residue was diluted with H ₂ 0 (20 mL), and adjusted to pH of 3-4 by 6.0 N hydrochloric acid. The mixture was extracted with EtOAc (30 mL x 3), washed with brine (15 mL), dried over Na ₂ S0 ₄ , filtered, and concentrated to give the crude title compound 4-(2-methyl-5-(2-methylpyridin-4-yl)-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[2,3-b]pyridin-3-yl )-lH-pyrrole-2-carboxylic acid

(XXVII) as a white solid (105 mg, crude yield >100%), which was used in the next step without any further purification.

Step 6. Preparation of (S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(2-methyl-5-(2- methylpyridin-4-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrrolo[2,3-b]pyridin-3-yl)- lH-pyrrole-2-carboxamide (XXIX)

[091] A solution of 4-(2-methyl-5-(2-methylpyridin-4-yl)-l-((2-(trimethylsilyl)e thoxy)methyl)- lH-pyrrolo[2,3-b]pyridin-3-yl)-lH-pyrrole-2-carboxylic acid (XXVII) (105 mg, 0.227 mmol), 2- amino-2-(3-chlorophenyl)ethanol (XXVIII) (47 mg, 0.272 mmol), HATU (129 mg, 0.340 mmol), and DIEA (88 mg, 0.681 mol) in DMF (5 mL) was stirred at the room temperature overnight. The mixture was diluted with EtOAc (50 mL), washed with H20 (15 mL x 2) and brine (20 mL), dried over Na ₂ S0 ₄ , filtered, and concentrated. The residue was purified by prep- TLC (DCM/MeOH=20/l) to give the title compound (S)-N-(l-(3-chlorophenyl)-2- hydroxyethyl)-4-(2-methyl-5-(2-methylpyridin-4-yl)- 1 -((2-(trimethylsilyl)ethoxy)methyl)- 1 H- pyrrolo[2,3-b]pyridin-3-yl)-lH-pyrrole-2-carboxamide (XXIX) as a yellow solid (45 mg, yield: 32%).

Step 7. Preparation of (S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(2-methyl-5-(2- methylpyridin-4-yl)-l -carboxamide (XXX)

[092] To a solution of (S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(2-methyl-5- (2- methylpyridin-4-yl)- 1 -((2-(trimethylsilyl)ethoxy)methyl)- 1 H-pyrrolo[2,3-b]pyridin-3-yl)- 1H- pyrrole-2-carboxamide (XXIX) (45 mg, 0.073 mmol) in 1 M TBAF/THF (3.0 mL) was added ethane- 1 ,2-diamine (131 mg, 2.191 mmol). The mixture was stirred at 90 °C overnight, diluted with EtOAc (50 mL), washed with H ₂ 0 (15 mL x 4) and brine (15 mL), dried over Na ₂ S0 ₄ , and concentrated. The residue was purified by column chromatography (DCM: MeOH = 10: 1) to give the title compound (S)-N-(l-(3-chlorophenyl)-2-hydroxyethyl)-4-(2-methyl-5-(2- methylpyridin-4-yl)-lH-pyrrolo[2,3-b]pyridin-3-yl)-lH-pyrrol e-2-carboxamide (XXX) as a yellow solid (12 mg, yield: 35%).

[093] Table 3 below lists some exemplary compounds of this invention that were synthesized accordingly to Scheme C.

Table 3

Example 4: Colo205 Cell Proliferation Assay

[094] Compounds of this invention were tested for the inhibition of ERK2 by a Colo205 cell proliferation assay commonly known as MTT assay. In this assay, a complete media is prepared by adding 10% fetal bovine serum to RPMI-1640 medium (Life technology). Colon cancer cells (Colo205 cell line) were added to each of 88 wells of a 96 well plate at a seeding density of 5,000 The cells were allowed to attach to the plate by incubating at 37°C for 24 hours. The compound was dissolved in DMSO (SIGMA). A solution of test compound was prepared in complete media by serial dilution to obtain the following concentrations: 500μΜ, 150μΜ, 50μΜ, 15μΜ, 5μΜ, 1.5μΜ, 0.5μΜ, 0.15μΜ and 0.05μΜ. The test compound solution (ΙΟμί) is added to each of 80 cell-containing wells. The final concentrations of the compound were following: 50μΜ, 15μΜ, 5μΜ, 1.5μΜ, 0.5μΜ, 0.15μΜ, 0.05μΜ, 0.015μΜ and 0.005μΜ. The final concentration of DMSO is 0.5%. To the 8 remaining cell-containing wells, only complete media (containing 0.5% DMSO) is added to form a control group in order to measure maximal proliferation. To the remaining 8 empty wells, complete media is added to for a vehicle control group in order to measure background. The plates are incubated at 37°C for 72 hours. ΙΟμί WST-8 solution (DOJI DO, Cell Counting KIT-8) was added to each well. The plates were further incubated at 37 °C for 2 hours, and then read for the absorbance using a microplate reader at 450 nm.

Example 5: Enzymatic Assay

[095] Compounds were tested in a LanthaScreen™ time-resolved fluorescence energy transfer (TR-FRET) enzymatic assay from Invitrogen. The assay used human ERK2 (Mitogen Activated Kinase 1 , Invitrogen, Cat. PV3311) recombinantly expressed as GST-tagged full-length protein purified from E. coli and activated in vitro with MAP2K1. The substrate was a recombinant truncated version (residues 19-96) of ATF2 fused with Green Fluorescent Protein (Invitrogen, Cat. PV4445). Test compounds were prepared and diluted in DMSO in 3-fold serial dilutions to 100X of the final testing concentrations. The compounds were then further diluted to 4X by the kinase reaction buffer (Invitrogen, Cat. PV3189). The enzymatic reaction for compound testing was performed in a white 384-well polypropylene plate (Packard, Cat. 6005214) with a total reaction volume of 10 μΐ containing 20 ng/ml ERK2, 400 nM substrate, and 5 μΜ ATP that is around its K _m . The assay started with loading 2.5 μΐ of ERK2 diluted in kinase reaction buffer to wells, followed by addition of equal volume of 4X compounds for 15-min incubation at the room temperature for pre-treatment. The enzymatic reaction was initiated by addition of 5 μΐ of mixture of the substrate and ATP prepared in kinase reaction buffer. After one hour reaction, 10 μΐ mixture of EDTA (final 10 mM) and terbium-labeled anti-pATF2 (pThr71 ) antibody (final 2 nM) (Invitrogen, Cat. PV4451 ) prepared in TR-FRET antibody dilution buffer (Invitrogen, Cat. PV3574) was added to stop the enzymatic reaction and produce TR-FRET signals. After 30 minutes of incubation at room temperature, the plate was read in Tecan Infinite F200 Pro with the following settings: Excitation 340 nm (30)/Emissionl 495 nm ( 10)/Emission2 520 nm (25). The TR-FRET values were dimensionless numbers that were calculated as the ratio of the acceptor (Green Fluorescent Protein) signal to the donor (Terbium) signal. Percent of control was calculated as the percentage of compound-treated vs 1% DMSO vehicle-treated. The dose- response curves were generated and the IC50S were calculated by nonlinear sigmoid curve fitting using GraphPad Prism.

[096] The growth inhibition activities of the disclosed compounds are shown in Table 4 below. In this table, the letter "A" represents an IC ₅₀ value within the range of 0 to 1 (including 1), the letter "B" represents an IC ₅₀ value within the range of 1 to 10 (including 10) nM; the letter "C" represents an IC ₅₀ value within the range of 10 to 50 (including 50) nM; the letter "D" represents an IC ₅₀ value within the range of 50 to 200 (including 200) nM; the letter "E" represents an IC ₅₀ value within the range of 200 to 750 (including 750) nM; the letter "F" represents an IC ₅₀ value within the range of over 1000 nM; the letter "G" represents an IC ₅₀ lower than 0.1 μΜ; the letter "H" represents an IC ₅₀ within the range of 0.1 to 0.5 (including 0.5) μΜ; the letter "I" represents an IC ₅₀ within the range of 0.5 to 1.0 (including 1.0) μΜ; the letter "J" represents an IC ₅₀ within the range of 1.0 to 5.0 (including 5.0) μΜ; the letter "K" represents an IC ₅₀ within the range of 5.0 to 10.0 (including 10.0) μΜ; and the letter "L" represents an IC ₅₀ greater than 10.0 μΜ.

Table 4. In Vitro Assay Data