WO2018057588A1

WO2018057588A1 - Furo[3,2-b]pyridine compounds useful as inhibitors of the par-2 signaling pathway

Info

Publication number: WO2018057588A1
Application number: PCT/US2017/052446
Authority: WO
Inventors: Bingcan Liu; Stéphane DORICH; Mylène DE LÉSÉLEUC; Kristina Dupont-Gaudet; Clint Alwyn JAMES; Louis Vaillancourt; Marc-André BEAULIEU; Claudio Sturino
Original assignee: Vertex Pharmaceuticals Incorporated
Priority date: 2016-09-21
Filing date: 2017-09-20
Publication date: 2018-03-29

Abstract

Disclosed are chemical entities which are compounds of Formula (I): (I) and pharmaceutically acceptable salts thereof, wherein A, R^1, R², E, n and Z are as defined herein. These chemical entities are useful as inhibitors of the PAR-2 signaling pathway. These chemical entities and pharmaceutically acceptable compositions comprising such chemical entities can be employed for treating various diseases, disorders, and conditions.

Description

Furo[3,2-£]pyridine Compounds Useful as

Inhibitors of the PAR-2 Signaling Pathway

RELATED APPLICATIONS

[0001] The present application claims priority to United States Provisional Application No.

62/397,899 filed on September 21, 2016. The entire contents of this application are incorporated herein by reference.

BACKGROUND

[0002] Protease-Activated Receptors (PARs) are a family of G-protein coupled receptors (GPCRs) comprising PAR-1, 2, 3, and 4. PARs are typically activated when enzymes (such as thrombin or trypsin) proteolytically cleave a portion of their N-terminal sequence. This cleavage exposes a region of the N-terminal extracellular domain (called the "tethered ligand") which is believed to bind to residues contained within the second extracellular loop of the PAR receptors, resulting in the stabilization of an active conformation. Short synthetic peptides mimicking the tethered ligand sequence have been successfully used to activate all of the PAR receptors, except PAR-3.

[0003] PAR-2 is activated by several host and pathogen-derived serine proteases, including trypsin, mast cell tryptase, certain tissue kallikreins, and members of the coagulation cascade TF-FVIIa and FVa-FXa. Synthetic ligands such as SLIGKV-NH₂ can selectively activate human PAR-2, although modified PAR-2 synthetic agonists such as 2-fluoryl-LIGRLO-NH₂ have been reported to be more potent activators of this receptor.

[0004] PAR-2 has been shown to be an important receptor in mediating inflammation, pain and itch. For example, PAR-2 activation results in inflammatory cytokine and chemokine release from keratinocytes, endothelial cells and from human epithelial cell lines such as A549. Moreover, the administration of PAR-2 activating proteases and synthetic agonists in vivo induce inflammatory responses. In particular, several studies have shown that intraplantar administration of PAR-2 agonists in rodents results in an edema response that is dependent in part on neuronal PAR-2 activation.

[0005] Similar studies have implicated PAR-2 as a mediator of neurogenic inflammation, nociception and in transmission of pain. This is mediated in part by the activation of PAR-2 dependent signaling pathways in dorsal root ganglia, the release of neuropeptides from C-fibers in peripheral tissues and spinal cord and the potentiation of transient receptor potential vaniloid 1 and 4 receptors in sensory neurons.

[0006] Several studies have demonstrated a role for PAR-2 activation in pruritus. Both direct activation of PAR-2 on nerve endings and indirect effects of PAR-2 on resident cells including keratinocytes are thought to contribute to itch.

[0007] Further, both in vitro and in vivo studies have demonstrated a role for PAR-2 activation in tissue remodeling. First, activation of PAR-2 can promote fibroblast and myofibroblast proliferation, and the secretion of growth factors such as CTGF and extracellular matrix components including collagen. In addition, PAR-2 activation was shown to be implicated in cellular migration and activation of this pathway has recently been shown to promote tumor growth and metastasis.

[0008] Numerous studies relying on the use of PAR-2 deficient mice, blocking PAR-2 antibodies or PAR-2 antagonists such as GB88 revealed an important role for PAR-2 activation in the pathophysiology of a variety of diseases including asthma, chronic pain, rheumatoid arthritis, periodontitis, inflammatory bowel diseases, irritable bowel syndrome, skin diseases, cancer, fibrotic diseases and neurological disease (reviewed in Yau et al, Journal of Medicinal Chemistry, July 2013). Other studies have shown that diet-induced obesity, adipose inflammation, and metabolic dysfunction correlating with PAR-2 expression are attenuated by PAR-2 antagonism.

[0009] For all of these reasons, there is a need for the development of potent and selective inhibitors of the PAR-2 signaling pathway as novel therapeutic agents.

SUMMARY

[0010] The present invention relates to chemical entities, including compounds and pharmaceutically acceptable salts thereof, useful as inhibitors of the PAR-2 signaling pathway. The present invention also relates to pharmaceutically acceptable compositions comprising the chemical entities of this invention; methods of treating of various diseases, disorders, and conditions using the chemical entities of this invention; processes for preparing the chemical entities of this invention; intermediates for the preparation of the chemical entities of this invention; and methods of using the chemical entities in in vitro applications.

[0011] In some aspects, the present invention provides a chemical entity which is compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

each of R and R² independently is selected from -H, halo, -CH₃, and -CF₃, provided that at least one of ¹ and R² is not -H;

A is cyclohexyl in which each of R¹ and R² is 4-methyl, or

A is phenyl in which R¹ is at the 4-position and R² is at the 3-position;

n is 1 or 2;

E is -CH₂- or -C(O)-; and

Z is N-X or CH-X³;

X is R⁵, -C(0)R⁵ or -S(0)₂R⁵;

X³ is -(CR₂)_r-C(0)OR⁶, -(CR₂)_r-N(R)R⁶, -(CR₂)_r-C(0)N(R)R⁶ or -(CR₂)_r-C(0)N(R)S(0)₂R⁶;

each instance of r independently is 0, 1 or 2;

each of R⁵ and R⁶ independently is -(V)_a-Y; wherein

V is C_1-6 aliphatic wherein up to three carbon units of said C_1-6 aliphatic are optionally and

independently replaced with -0-, -N R-, -S-, -C(O)- or -S(0)₂-; wherein V is optionally substituted with 1-4 occurrences of J^v;

each instance of J independently is halogen, -C0₂H, -CN, haloCi_₄alkyl or Ci_₄alkyl, wherein up to one methylene unit of each of said C^alkyl and haloC^alkyl is optionally replaced with -0-, -NR-, -S- or -C(O)-;

Y is H; -CN; a 3-7 membered, saturated, partially unsaturated or aromatic, monocyclic ring having 0-4 heteroatoms independently selected from oxygen, nitrogen and sulfur; or a 6-10 membered, saturated, partially unsaturated or aromatic, bicyclic ring having 0-6 heteroatoms independently selected from oxygen, nitrogen and sulfur; wherein Y is optionally substituted with 1-4 occurrences of J^Y;

each instance of J^Y independently is -H; oxo; halogen; -C0₂H; -CN; phenyl; 5-6-membered

heteroaryl having 1-4 heteroatoms independently selected from oxygen, nitrogen and sulfur; or C_1-6 aliphatic, wherein up to three carbon units of said C_1-6 aliphatic are optionally and independently replaced with -0-, -N R-, -S-, -C(O)- or -S(0)₂-; and wherein each of the phenyl, 5-6 membered heteroaryl and the C_1-6aliphatic is optionally and independently substituted with 1-4 substituents independently selected from the group consisting of halogen, -CN,

-OP(0)(OH)₂, -P(0)( )(OH) and

each R independently is -H or Ci_₄ alkyl; and

a is 0 or 1;

provided that the compound of Formula I is not any of the excluded compounds listed herein.

[0012] In some aspects, the present invention provides a pharmaceutical composition comprising a chemical entity described herein and a pharmaceutically acceptable carrier, adjuvant, or excipient.

[0013] In some aspects, the present invention provides a method for treating a PAR-2-mediated disease in a patient comprising administering to the patient an effective amount of a chemical entity described herein.

[0014] In some aspects, the present invention provides a method for treating inflammation or nociception (pain) in a patient comprising administering to the patient an effective amount of a chemical entity described herein.

[0015] In some aspects, the present invention provides a method for treating inflammatory bowel disease, Crohn's disease, irritable bowel syndrome, ulcerative colitis, asthma, rheumatoid arthritis, osteoarthritis, fibrosis, gingivitis, atopic dermatitis, psoriasis, systemic lupus erythematosus (SLE), scleroderma, interstitial lung disease, polymyositis, periodontitis, vasculitis, Netherton syndrome, atopic dermatitis, dermatomyositis, uveitis, Alzheimer's disease, Parkinson's disease, multiple sclerosis, inflammatory pain, post-operative incision pain, neuropathic pain, fracture pain, osteroporotic fracture pain, gout joint pain, cancer, diet-induced obesity, adipose inflammation, and/or metabolic dysfunction correlating with PAR-2 expression in a patient comprising administering an effective amount of a chemical entity described herein.

[0016] In some aspects, the present invention provides a method of inhibiting proteolytic activation of PAR-2 in a cell comprising administering to a patient or to a biological sample an effective amount of a chemical entity described herein.

[0017] In some aspects, the present invention provides a method of inhibiting PAR-2 activity in a cell comprising administering to a patient or to a biological sample an effective amount of a of chemical entity described herein. [0018] In some aspects, the present invention includes methods of preparing the chemical entities of Formula (I).

DETAILED DESCRIPTION OF CERTAIN EM BODIM ENTS OF TH E I NVENTION

[0019] In some embodiments, the present invention provides a chemical entity (a "provided chemical entity") which is compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

each of R and R² independently is selected from -H, halo, -CH₃, and -CF₃, provided that at least one of R¹ and R² is not -H;

A is cyclohexyl in which each of R¹ and R² is 4-methyl, or

A is phenyl in which R¹ is at the 4-position and R² is at the 3-position;

n is 1 or 2;

E is -CH₂- or -C(O)-; and

Z is N-X or CH-X³;

X is R⁵, -C(0)R⁵ or -S(0)₂R⁵;

each instance of r independently is 0, 1 or 2;

each of R⁵ and R⁶ independently is -(V)_a-Y; wherein

each instance of J independently is halogen, -C0₂H, -CN, haloC^alkyl or C_1-4alkyl, wherein up to one methylene unit of each of said C^alkyl and haloC^alkyl is optionally replaced with -0-, -NR-, -S- or -C(O)-;

Y is H; -CN; a 3-7 membered, saturated, partially unsaturated or aromatic, monocyclic ring

having 0-4 heteroatoms independently selected from oxygen, nitrogen and sulfur; or a 6-10 membered, saturated, partially unsaturated or aromatic, bicyclic ring having 0-6 heteroatoms independently selected from oxygen, nitrogen and sulfur; wherein Y is

optionally substituted with 1-4 occurrences of J^Y;

each instance of J^Y independently is -H; oxo; halogen; -C0₂H; -CN; phenyl; 5-6-membered heteroaryl having 1-4 heteroatoms independently selected from oxygen, nitrogen and sulfur; or Ci__s aliphatic, wherein up to three carbon units of said Ci_-6 aliphatic are optionally and independently replaced with -0-, -NR-, -S-, -C(O)- or -S(0)₂-; and wherein each of the phenyl, 5-6 membered heteroaryl and the Ci_₆aliphatic is optionally and independently substituted with 1-4 substituents independently selected from the group consisting of halogen, -CN,

-OP(0)(OH)₂, -P(0)(R)(OH) and

each R independently is -H or C_H alkyl; and

a is 0 or 1;

provided that the compound of Formula I is not any of the following:

-7-

[0020] As used herein, the term "excluded compounds" refers to the foregoing compounds which, along with their pharmaceutically acceptable salts, are explicitly excluded from the chemical entities of the present invention.

[0021] In some embodiments, a provided chemical entity is a compound of Formula (II):

or a pharmaceutically acceptable salt thereof, wherein A, R¹, R² and X are as defined for Formula (I), above, and in embodiments of Formula (II), below, both singly and in combination. 2] In some embodiments, in a compound of Formula (II)—

-21-

-22-

w

herein G¹ is and G² is

[0023] In some embodiments, a provided chemical entity is a compound of Formula (III):

or a pharmaceutically acceptable salt thereof, wherein A, R¹, R² and X are as defined for Formula (I), above, and in embodiments of Formula (III), below, both singly and in combination.

[0024] In some embodiments, in a compound of Formula (III)—

[0025] In some embodiments, a provided chemical entity is a compound of Formula (IV):

or a pharmaceutically acceptable salt thereof, wherein A, R¹, R² and X³ are as defined for Formula (I), above, and in embodiments of Formula (IV), below, both singly and in combination.

[0026] In some embodiments, in a compound of Formula (IV)—

or X³ is -NH₂;

[0027] In some embodiments, a provided chemical entity is a compound of Formula (V):

or a pharmaceutically acceptable salt thereof, wherein A, R¹, R², E and Z are as defined for Formula (I), above, and in embodiments of Formula (V), below, both singly and in combination.

[0028] In some embodiments, in a compound of Formula (V)—

E is -CH₂- or -C(O)-;

or E is -CH₂-;

or E is -C(O)-; and

9] In some embodiments, a provided chemical entity is a compound of Formula (VI):

or a pharmaceutically acceptable salt thereof, wherein A, ¹, R², n and E are as defined for Formula (I), above, and in embodiments of Formula (VI), below, both singly and in combination, and wherein:

X⁴ is N;

X⁵ is N, CH or CR¹⁰, wherein

R¹⁰ is halogen; -C0₂H; -CN; phenyl; 5-6-membered heteroaryl having 1-4 heteroatoms

independently selected from oxygen, nitrogen and sulfur; or Ci_₆ aliphatic, wherein up to three carbon units of said Ci_₆ aliphatic are optionally and independently replaced with -0-, - NR-, -S-, -C(O)- or -S(0)₂-; and wherein each of the phenyl, 5-6 membered heteroaryl and the Ci_6 aliphatic is optionally and independently substituted with 1-4 substituents independently selected from the group consisting of halogen, -CN,

-OP(0)(OH)₂, -P(0)(R)(OH) and

each of R⁷ , R⁸ and R⁹ independently is selected from -H, halo, C_1-4 alkyl, C_1-4 haloalkyl, -OH, -OR^J6, -NH₂, -N HR^J6, -N(R^J6)₂, -C(0)R^J6, -C0₂H, -C(0)OR^J6, -C(0)NH₂, -C(0)NH R^J6, -C(0)N(R^J6)₂, -S(0)₂NH₂, -S(0)₂N HR^J6, -S(0)₂N(R^J6)₂, -C(0)N(R^J6)S(0)₂R^J6, -C(0)N(R^J6)S(0)₂N HR^J6,

-C(0)N(R^J6)S(0)₂N(R^J6)₂, -CN and tetrazolyl, wherein said tetrazolyl is unsubstituted or substituted with R^J6;

provided that at least one of R⁷ , R⁸ and R⁹ is not -H,

wherein each instance of R^J6 independently is Ci_₃ alkyl. 0] In some embodiments, in a compound of Formula (VI)—

n is 1 or 2;

or n is 1;

or n is 2;

E is -CH₂- or -C(O)-;

or E is -CH₂-;

or E is -C(O)-;

X⁵ is N, CH or C ¹⁰;

or X⁵ is N or CH;

or X⁵ is N;

or X⁵ is CH;

R⁷ is -H, halo, d_₄ alkyl, -OH, -OR^JS, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^JS, -CN or unsubstituted tetrazolyl;

or R^7' is -H, -F, -CI, -CH₃, Et, -OH, -OMe, -NH₂, -NHMe, -C0₂H, -C(0)OMe, -C(0)OEt, -CN or unsubstituted tetrazolyl;

or R^7' is -H, -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl;

R⁸ is -H, halo, d_₄ alkyl, -OH, -OR^JS, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^JS, -CN or unsubstituted tetrazolyl;

or R⁸ is -H, -F, -CI, -CH₃, Et, -OH, -OMe, -NH₂, -NHMe, -C0₂H, -C(0)OMe, -C(0)OEt,-CN or unsubstituted tetrazolyl;

or R^8' is -H, -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl;

R^9' is -H, halo, C₁₄ alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6,-CN or unsubstituted tetrazolyl; or R^9' is -H, -F, -CI, -CH₃, Et, -OH, -OMe, -NH₂, -NHMe, -C0₂H, -C(0)OMe, -C(0)OEt,-CN or unsubstituted tetrazolyl;

or ⁹ is -H, -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl; and

R¹⁰ is halo, C_w alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6, -CN or unsubstituted tetrazolyl; or R¹⁰ is -F, -CI, -CH₃, Et, -OH, -OMe, -NH₂, -NHMe, -C0₂H, -C(0)OMe, -C(0)OEt,-CN or

unsubstituted tetrazolyl;

or R¹⁰ is -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl. 1] In some embodiments, in a compound of Formula (VI)—

n is 1; E is -CH₂-; X⁵ is CH;

R^7' is -H, halo, d_-4 alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6, -CN or unsubstituted

tetrazolyl;

or R^7' is -H, -F, -CI, -CH₃, Et, -OH, -OMe, -NH₂, -NHMe, -C0₂H, -C(0)OMe, -C(0)OEt,-CN or

unsubstituted tetrazolyl;

or R^7' is -H, -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl;

R^8' is -H, halo, C_1-4 alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6, -CN or unsubstituted

tetrazolyl; or R^8' is -H, -F, -CI, -CH₃, Et, -OH, -OMe, -NH₂, -NHMe, -C0₂H, -C(0)OMe, -C(0)OEt,-CN or unsubstituted tetrazolyl;

or ⁸ is -H, -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl; and

R⁹ is -H, halo, C_1-4 alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6, -CN or unsubstituted

tetrazolyl;

or R^9' is -H, -F, -CI, -CH₃, Et, -OH, -OMe, -NH₂, -NHMe, -C0₂H, -C(0)OMe, -C(0)OEt,-CN or

unsubstituted tetrazolyl;

or R^9' is -H, -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl. 2] In some embodiments, a provided chemical entity is a compound of Formula (VI.A):

or a pharmaceutically acceptable salt thereof, wherein A, R¹, R², n and E are as defined for Formula (I), above, and in embodiments of Formula (VI), below, both singly and in combination, and wherein:

X⁴ is N;

X⁵ is N, CH or CR^10', wherein

independently selected from oxygen, nitrogen and sulfur; or C_1-6 aliphatic, wherein up to three carbon units of said C_1-6 aliphatic are optionally and independently replaced with -0-, - NR-, -S-, -C(O)- or -S(0)₂-; and wherein each of the phenyl, 5-6 membered heteroaryl and the C_1-6aliphatic is optionally and independently substituted with 1-4 substituents independently selected from the group consisting of halogen, -CN,

R⁸ and R⁹ independently is selected from halo, C_1-4 alkyl, C_1-4 haloalkyl, -OH, -OR NHR^J6, -N(R^J6)₂, -C(0)R^J6, -C0₂H, -C(0)OR^J6, -C(0)NH₂, -C(0)NHR^J6, -C(0)N(R^J6)₂, -S(0)₂NH₂, -S(0)₂NH ^J6, -S(0)₂N(R^J6)₂, -C(0)N(R^J6)S(0)₂R^J6, -C(0)N(R^J6)S(0)₂NHR^J6, -C(0)N(R^J6)S(0)₂N(R^J6)₂, -CN and tetrazolyl, wherein said tetrazolyl is unsubstituted or substituted with R^J6;

wherein each instance of R^J6 independently is C_1-3 alkyl. 3] In some embodiments, in a compound of Formula (VI.A)—

n is 1 or 2;

or n is 1;

or n is 2;

E is -CH₂- or -C(O)-;

or E is -CH₂-;

or E is -C(O)-;

X⁵ is N, CH or CR¹⁰;

or X⁵ is N or CH;

or X⁵ is N;

or X⁵ is CH; R⁷ is halo, d_-4 alkyl, -OH, -O ^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6, -CN or unsubstituted tetrazolyl; or R⁷ is -F, -CI, -CH₃, Et, -OH, -OMe, -NH₂, -NHMe, -C0₂H, -C(0)OMe, -C(0)OEt, -CN or

unsubstituted tetrazolyl;

or R⁷ is -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl;

or R⁷ is -F, -CI, -CH₃, -C0₂H or -CN;

R⁸ is halo, C₁₄ alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6, -CN or unsubstituted tetrazolyl; or R⁸ is -F, -CI, -CH₃, Et, -OH, -OMe, -NH₂, -NHMe, -C0₂H, -C(0)OMe, -C(0)OEt,-CN or

unsubstituted tetrazolyl;

or R⁸ is -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl;

or R⁸ is -F, -CI, -CH₃, -C0₂H or -CN;

R⁹ is halo, C₁₄ alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6,-CN or unsubstituted tetrazolyl; or R⁹ is -F, -CI, -CH₃, Et, -OH, -OMe, -NH₂, -NHMe, -C0₂H, -C(0)OMe, -C(0)OEt,-CN or

unsubstituted tetrazolyl;

or R⁹ is -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl;

or R⁹ is -F, -CI, -CH₃, -C0₂H or -CN; and

R¹⁰ is halo, C_M alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6, -CN or unsubstituted tetrazolyl; or R¹⁰ is -F, -CI, -CH₃, Et, -OH, -OMe, -NH₂, -NHMe, -C0₂H, -C(0)OMe, -C(0)OEt,-CN or

unsubstituted tetrazolyl;

or R¹⁰ is -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl;

or R¹⁰ is -F, -CI, -CH₃, -C0₂H or -CN. 4] In some embodiments, in a compound of Formula (VI.A)—

n is 1;

E is -CH ; X⁵ is CH;

R⁷ is halo, C_1-4 alkyl, -OH, -O ^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6, -CN or unsubstituted tetrazolyl; or R⁷ is -F, -CI, -CH₃, Et, -OH, -OMe, -NH₂, -NHMe, -C0₂H, -C(0)OMe, -C(0)OEt, -CN or

unsubstituted tetrazolyl;

or R⁷ is -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl;

or R⁷ is -F, -CI, -CH₃, -C0₂H or -CN;

or R⁷ is -CH₃, -C0₂H or -CN;

or R⁷ is -CH₃ or -C0₂H;

unsubstituted tetrazolyl;

or R⁸ is -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl;

or R⁸ is -F, -CI, -CH₃, -C0₂H or -CN;

or R⁸ is -CH₃, -C0₂H or -CN;

or R⁸ is -CH₃ or -C0₂H; and

R⁹ is halo, C_1-4 alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6,-CN or unsubstituted tetrazolyl; or R⁹ is -F, -CI, -CH₃, Et, -OH, -OMe, -NH₂, -NHMe, -C0₂H, -C(0)OMe, -C(0)OEt,-CN or

unsubstituted tetrazolyl;

or R⁹ is -F, -CI, -CH₃, -C0₂H, -CN or unsubstituted tetrazolyl;

or R⁹ is -F, -CI, -CH₃, -C0₂H or -CN;

or R⁹ is -CH₃, -C0₂H or -CN;

or R⁹ is -CH₃ or -C0₂H. 5] In some embodiments, in a compound of Formula (VI.A)—

n is 1;

E is -CH X⁵ is CH; and 7 is -CH₃, R⁸ is -CH₃ and R⁹ is -C0₂H;

or R⁷ is -CH₃, R⁸ is -C0₂H and R⁹ is -CH₃;

or R⁷ is -C0₂H, R⁸ is -CH₃ and R⁹ is -CH₃.

[0036] In some embodiments, in a compound of Formula (VI.A)—

n is 1;

E is -CH₂-; X⁵ is CH; and

R⁷ is -CH₃, R⁸ is -CH₃ and R⁹ is -C0₂H;

or R⁷ is -CH₃, R⁸ is -C0₂H and R⁹ is -CH₃;

or R⁷ is -C0₂H, R⁸ is -CH₃ and R⁹ is -CH₃.

[0037] In some embodiments, in a compound of Formula (VI.A)—

n is 1;

E is -CH₂-; X⁵ is CH; and

R⁷ is -CH₃, R⁸ is -CH₃ and R⁹ is -C0₂H;

or R⁷ is -CH₃, R⁸ is -C0₂H and R⁹ is -CH₃;

or R⁷ is -C0₂H, R⁸ is -CH₃ and R⁹ is -CH₃.

Definitions

[0038] As used herein, the term "including" and other forms thereof such as "include", "includes", etc. are intended to be open-ended unless otherwise specified or clear from context. That is, "including" is to be understood as "including but not limited to" unless otherwise specified or clear from context.

[0039] As used herein, the term "chemical entity" refers to a compound described herein, generically or specifically, or a pharmaceutically acceptable salt thereof. The term "free compound", or "free acid" or "free base", as applicable, refers to the compound as described. Statements herein regarding "compounds" apply equally to chemical entities and, as applicable, vice-versa. Accordingly, no significance is intended by the use of "chemical entity" in some contexts and "compound" in others, with respect to the description of the compound.

[0040] Compounds of this invention include those described generally herein, and are further illustrated by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, HANDBOOK OF CHEMISTRY AND PHYSICS, 75^TH Ed. Additionally, general principles of organic chemistry are described in M. Loudon and J. Parise, ORGANIC CHEMISTRY, 6^TH Ed., W.H. Freeman & Co.: New York (2016), and M .B. Smith, MARCH'S ADVANCED ORGANIC CHEMISTRY, 7^TH Ed., John Wiley & Sons, Inc.: Hoboken (2013), the entire contents of each of which are hereby incorporated by reference.

[0041] As described herein, a specified number range of atoms includes any integer therein. For example, a group having from 1-4 atoms could have 1, 2, 3, or 4 atoms.

[0042] As described herein, compounds of the invention may optionally be substituted with one or more substituents, such as are illustrated generally herein, or as exemplified by particular classes, subclasses, and species of the invention. It will be appreciated that the phrase "optionally substituted" is used interchangeably with the phrase "substituted or unsubstituted." In general, the term "substituted", whether preceded by the term "optionally" or not, refers to the replacement of hydrogen radicals in a given structure with the radical of a specified substituent. Unless otherwise indicated, an optionally substituted group may have a substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this invention are preferably those that result in the formation of stable or chemically feasible compounds. [0043] Unless otherwise indicated, a substituent connected by a bond drawn from the center of a ring means that the substituent can be bonded to any position in the ring. In example (i) below, for instance, J¹ can be bonded to any position on the pyridyl ring. For bicyclic rings, a bond drawn through both rings indicates that the substituent can be bonded from any position of the bicyclic ring. In example (ii) below, for instance, J¹ can be bonded to the 5-membered ring (on the nitrogen atom, for instance), and to the 6-membered ring.

(") (ii)

[0044] The term "stable", as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, recovery, purification, and use for one or more of the purposes disclosed herein. In some embodiments, a stable compound or chemically feasible compound is one that is not substantially altered when kept at a temperature of 40°C or less, in the absence of moisture or other chemically reactive conditions, for at least a week.

[0045] The term "aliphatic" or "aliphatic group", as used herein, means a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted, hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation that has a single point of attachment to the rest of the molecule.

[0046] Unless otherwise specified, aliphatic groups contain 1-20 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-10 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-8 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-6 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. Aliphatic groups may be linear or branched, substituted or unsubstituted alkyl, alkenyl, or alkynyl groups. Specific examples include methyl, ethyl, isopropyl, n-propyl, sec-butyl, vinyl, n-butenyl, ethynyl, and ferf-butyl.

[0047] The term "cycloaliphatic" (or "carbocycle" or "carbocyclyl") refers to a monocyclic C₃-C₈ hydrocarbon or bicyclic C₈-Ci₂ hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of attachment to the rest of the molecule wherein any individual ring in said bicyclic ring system has 3-7 members. Examples of cycloaliphatic groups include cycloalkyl and cycloalkenyl groups. Specific examples include, cyclohexyl, cyclopropenyl, and cyclobutyl.

[0048] The term "heterocycle", "heterocyclyl", or "heterocyclic" as used herein means non- aromatic, monocyclic, bicyclic, or tricyclic ring systems in which one or more ring members are an independently selected heteroatom. In some embodiments, the "heterocycle", "heterocyclyl", or "heterocyclic" group has three to fourteen ring members in which one or more ring members is a heteroatom independently selected from oxygen, sulfur, nitrogen, or phosphorus, and each ring in the system contains 3 to 7 ring members.

[0049] Examples of heterocycles include 3-lH-benzimidazol-2-one, 3-(l-alkyl)-benzimidazol-2-one, 2-tetrahydrofuranyl, 3-tetrahydrofuranyl, 2-tetrahydrothiophenyl, 3-tetrahydrothiophenyl,

2- morpholino, 3-morpholino, 4-morpholino, 2-thiomorpholino, 3-thiomorpholino, 4-thio- morpholino, 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, 1-tetrahydropiperazinyl, 2-tetra- hydropiperazinyl, 3-tetrahydropiperazinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 1- pyrazolinyl,

3- pyrazolinyl, 4-pyrazolinyl, 5-pyrazolinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 2-thiazolidinyl, 3-thiazolidinyl, 4-thiazolidinyl, 1-imidazolidinyl, 2-imidazolidinyl, 4-imidazolidinyl, 5-imidazolidinyl, indolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzothiolane, benzodithiane, and l,3-dihydro-imidazol-2-one.

[0050] Cyclic groups, (e.g. cycloaliphatic and heterocycles), can be linearly fused, bridged, or spirocyclic.

[0051] The term "heteroatom" means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including, any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quaternized form of any basic nitrogen or; a substitutable nitrogen of a heterocyclic ring, for example N (as in 3,4-dihydro-2/-/-pyrrolyl), NH (as in pyrrolidinyl) or N ⁺ (as in N-substituted pyrrolidinyl)).

[0052] The term "unsaturated", as used herein, means that a moiety has one or more units of unsaturation. Examples of unsaturated groups include, propyne, butene, cyclohexene, tetrahydropyridine and cyclooctatetraene. The term "alkoxy", or "thioalkyl", as used herein, refers to an alkyl group, as previously defined, attached through an oxygen ("alkoxy") or sulfur ("thioalkyl") atom. [0053] The terms "haloalkyl" (e.g., haloC^alkyl), "haloalkenyl", "haloaliphatic", and "haloalkoxy" mean alkyl, alkenyl or alkoxy, as the case may be, substituted with one or more halogen atoms. This term includes perfluorinated alkyl groups, such as -CF₃ and -CF₂CF₃.

[0054] The terms "halogen", "halo", and "hal" mean F, CI, Br, or I.

[0055] The term "aryl" used alone or as part of a larger moiety as in "aralkyl", "aralkoxy", or "aryloxyalkyl", refers to carbocyclic aromatic ring systems. The term includes monocyclic, bicyclic, and tricyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 7 ring members. The term "aryl" may be used interchangeably with the term "aryl ring".

[0056] The term "heteroaryl", used alone or as part of a larger moiety as in "heteroaralkyl" or "heteroarylalkoxy", refers to monocyclic, bicyclic, and tricyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic, at least one ring in the system contains one or more heteroatoms, and wherein each ring in the system contains 3 to 7 ring members. The term "heteroaryl" may be used interchangeably with the term "heteroaryl ring" or the term "heteroaromatic". Examples of heteroaryl rings include 2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, benzimidazolyl, 3-isoxazolyl, 4- isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2- pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (e.g., 3- pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (e.g., 5-tetrazolyl), triazolyl (e.g., 2- triazolyl and 5-triazolyl), 2-thienyl, 3-thienyl, benzofuryl, benzothiophenyl, indolyl (e.g., 2-indolyl), pyrazolyl (e.g., 2-pyrazolyl), isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, purinyl, pyrazinyl, 1,3,5-triazinyl, quinolinyl (e.g., 2-quinolinyl, 3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g., 1-isoquinolinyl, 3-isoquinolinyl, or 4-isoquinolinyl).

[0057] It should be understood that the term "heteroaryl" includes certain types of heteroaryl rings that exist in equilibrium between two different forms. More specifically, for example, species such hydropyridine and pyridinone (and likewise hydroxypyrimidine and pyrimidinone) are meant to be encompassed within the definition of "heteroaryl."

OH O [0058] The terms "protecting group" and "protective group" as used herein, are interchangeable and refer to an agent used to temporarily block one or more desired functional groups in a compound with multiple reactive sites. In certain embodiments, a protecting group has one or more, or preferably all, of the following characteristics: a) is added selectively to a functional group in good yield to give a protected substrate that is b) stable to reactions occurring at one or more of the other reactive sites; and c) is selectively removable in good yield by reagents that do not attack the regenerated, deprotected functional group. As would be understood by one skilled in the art, in some cases, the reagents do not attack other reactive groups in the compound. In other cases, the reagents may also react with other reactive groups in the compound. Examples of protecting groups are detailed in Greene, T.W., Wuts, P. G in " Protective Groups in Organic Synthesis", Third Edition, John Wiley & Sons, New York: 1999 ("Greene") (and other editions of the book), the entire contents of which are hereby incorporated by reference. The term "nitrogen protecting group", as used herein, refers to an agent used to temporarily block one or more desired nitrogen reactive sites in a multifunctional compound. Preferred nitrogen protecting groups also possess the characteristics exemplified for a protecting group above, and certain exemplary nitrogen protecting groups are also detailed in Chapter 7 in Greene.

[0059] In some embodiments, a methylene or carbon unit of an alkyl or aliphatic chain is optionally replaced with another atom or group. Examples of such atoms or groups include nitrogen, oxygen, sulfur, -C(O)-, -C(=N-CN)-, -C(=N )-, -C(=NOR)-, -SO-, and -S0₂-. These atoms or groups can be combined to form larger groups. Examples of such larger groups include -OC(O)- , -C(0)CO-, -CO , -C(0)NR-, -C(=N-CN), -N RCO-, -N RC(0)0-, -S0₂N R-, -NRSO , -NRC(0)N R- , -OC(0)NR-, and -N RS0₂NR-, wherein R is, for example, H or C_1-6 aliphatic. It should be understood that these groups can be bonded to the methylene or carbon units of the aliphatic chain via single, double, or triple bonds. An example of an optional replacement (nitrogen atom in this case) that is bonded to the aliphatic chain via a double bond would be -CH₂CH=N-CH₃. In some cases, especially on the terminal end, an optional replacement can be bonded to the aliphatic group via a triple bond. One example of this would be CH₂CH₂CH₂C≡N. It should be understood that in this situation, the terminal nitrogen is not bonded to another atom.

[0060] It should also be understood that, the term "methylene unit" or "carbon unit" can also refer to branched or substituted methylene or carbon units. For example, in an isopropyl moiety [-CH(CH₃)₂], a nitrogen atom (e.g., N R) replacing the first recited "methylene unit" would result in dimethylamine [-N(CH₃)₂]. In instances such as these, one of skill in the art would understand that the nitrogen atom will not have any additional atoms bonded to it, and the " " from "NR" would be absent in this case.

[0061] Unless otherwise indicated, the optional replacements form a chemically stable compound.

Optional replacements can occur both within the chain and/or at either end of the chain; i.e. both at the point of attachment and/or also at the terminal end. Two optional replacements can also be adjacent to each other within a chain so long as it results in a chemically stable compound. For example, a C₃ aliphatic can be optionally replaced by 2 nitrogen atoms to form - C-N≡N. The optional replacements can also completely replace all of the carbon atoms in a chain. For example, a C₃ aliphatic can be optionally replaced by -NR-, -C(O)-, and -NR- to form -NRC(0)NR- (a urea).

[0062] Unless otherwise indicated, if the replacement occurs at the terminal end, the replacement atom is bound to a hydrogen atom on the terminal end. For example, if a methylene unit of -CH2CH2CH3 were optionally replaced with -0-, the resulting compound could be -OCH2CH3, -CH2OCH3, or -CH2CH2OH. It should be understood that if the terminal atom does not contain any free valence electrons, then a hydrogen atom is not required at the terminal end (e.g., -CH₂CH₂CH=0 or -CH₂CH₂C≡N).

[0063] Unless otherwise indicated, structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, geometric, conformational, and rotational) forms of the structure. For example, the R and S configurations for each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers are included in this invention. As would be understood to one skilled in the art, a substituent can freel rotate around any

rotatable bonds. For example, a substituent drawn as

[0064] Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, geometric, conformational, and rotational mixtures of the present compounds are within the scope of the invention.

[0065] Unless otherwise indicated, all tautomeric forms of the compounds of the invention are within the scope of the invention. [0066] In some aspects, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a ¹³C- or ¹⁴C-enriched carbon are within the scope of this invention. Such compounds are useful, for example, for therapeutics and/or analytical tools or probes in biological assays. Especially deuterium (²H)-labeled compounds can also be used for therapeutic purposes.

[0067] In some embodiments, the invention is directed to isotope-labeled chemical entities, which are isotope-labeled compounds of Formula (Ι'), or pharmaceutically acceptable salts thereof, wherein the formula and variables of Formula (Γ) are each and independently as described above for Formula (I) or any other embodiments described above, provided that one or more atoms therein have been replaced by an atom or atoms having an atomic mass or mass number which differs from the atomic mass or mass number of the atom which usually occurs naturally. Examples of isotopes which are commercially available and suitable for the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, for example,

2_U H, 3_U H, 13, C-, I C, 15 _M N, 18 O, 17 O, 31_n P, 32 _n P, 35_c S, 18_c F and 36_r C,I, respectively.

[0068] The isotope-labeled chemical entities of the invention can be used in a number of beneficial ways. They can be suitable for medicaments and/or various types of assays, such as substrate tissue distribution assays. For example, tritium (³H)- and/or carbon-14 (¹⁴C)-labeled compounds are particularly useful for various types of assays, such as substrate tissue distribution assays, due to relatively simple preparation and excellent detectability. For example, deuterium (²H)- labelled compounds are therapeutically useful with potential therapeutic advantages over the non-²H-labelled compounds. In general, deuterium (²H)-labeled compounds can have higher metabolic stability as compared to those compounds that are not isotope-labeled owing to the kinetic isotope effect described below. Higher metabolic stability generally translates directly into an increased in vivo half-life or lower dosages, which under most circumstances would represent a preferred embodiment of the present invention. The isotope-labeled compounds of the invention can usually be prepared by carrying out the procedures described herein, replacing a non-isotope-labeled reactant by a readily available isotope-labeled reactant.

[0069] In some embodiments, the isotope-labeled compounds of the invention are deuterium (²H)- labeled compounds. In some embodiments, the invention is directed to deuterium (²H)-labeled chemical entities of Formula (Γ). In some embodiments, one, two, three or four hydrogen atoms are replaced by deuterium. [0070] Deuterium (²H)-labeled compounds of the invention can manipulate the oxidative metabolism of the compound by way of the primary kinetic isotope effect. The primary kinetic isotope effect is a change of the rate for a chemical reaction that results from exchange of isotopic nuclei, which in turn is caused by the change in ground state energies necessary for covalent bond formation after this isotopic exchange. Exchange of a heavier isotope usually results in a lowering of the ground state energy for a chemical bond and thus causes a reduction in the rate-limiting bond breakage. If the bond breakage occurs in or in the vicinity of a saddle- point region along the coordinate of a multi-product reaction, the product distribution ratios can be altered substantially. For explanation: if deuterium is bonded to a carbon atom at a non- exchangeable position, rate differences of k_M/k_D = 2-7 are typical. If this rate difference is successfully applied to, for example, a compound of Formula (Ι'), the profile of this compound in vivo can be drastically modified and result in improved pharmacokinetic properties. For a further discussion, see S. L. Harbeson and . D. Tung, Deuterium In Drug Discovery and Development, Ann. Rep. Med. Chem. 2011, 46, 403-417, incorporated in its entirety herein by reference.

[0071] The concentration of the isotope(s) (e.g., deuterium) incorporated into the isotope-labelled compounds of the invention may be defined by the isotopic enrichment factor. The term "isotopic enrichment factor" as used herein means the ratio between the isotopic abundance and the natural abundance of a specified isotope. In some embodiments, if a substituent in a compound of the invention is denoted deuterium, such compound has an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation).

[0072] When discovering and developing therapeutic agents, the person skilled in the art attempts to optimize pharmacokinetic parameters while retaining desirable in vitro properties. It may be reasonable to assume that many compounds with poor pharmacokinetic profiles are susceptible to oxidative metabolism. In vitro liver microsomal assays currently available provide valuable information on the course of oxidative metabolism of this type, which in turn permits the rational design of the deuterium (²H)-labeled compounds of the invention which can have improved stability through resistance to such oxidative metabolism. Significant improvements in the pharmacokinetic profiles of such compounds can thereby be obtained, and can be expressed quantitatively in terms of increases in the in vivo half-life (t^), concentration at maximum therapeutic effect (C_max), area under the dose response curve (AUC), and bioavailability; and in terms of reduced clearance, dose and materials costs.

[0073] The following is intended to illustrate the above: a deuterium (²H)-labeled compound of the invention, which has multiple potential sites of attack for oxidative metabolism, for example benzylic hydrogen atoms and hydrogen atoms bonded to a nitrogen atom, is prepared as a series of analogues in which various combinations of hydrogen atoms are replaced by deuterium atoms, so that some, most or all of these hydrogen atoms have been replaced by deuterium atoms. Half-life determinations enable favorable and accurate determination of the extent to which the improvement in resistance to oxidative metabolism has improved. In this way, it is determined that the half-life of the parent compound can be extended by up to 100% as the result of deuterium-hydrogen exchange of this type.

[0074] Deuterium-hydrogen exchange in a deuterium (²H)-labeled compound of the invention can also be used to achieve a favorable modification of the metabolite spectrum of the starting compound in order to diminish or eliminate undesired toxic metabolites. For example, if a toxic metabolite arises through oxidative carbon-hydrogen (C-H) bond cleavage, it can reasonably be assumed that the deuterated analogue will greatly diminish or eliminate production of the unwanted metabolite, even if the particular oxidation is not a rate-determining step. Further information on the state of the art with respect to deuterium-hydrogen exchange may be found, for example in Hanzlik et al., J. Org. Chem. 55, 3992-3997, 1990, eider et al., J. Org. Chem. 52, 3326-3334, 1987, Foster, Adv. Drug Res. 14, 1-40, 1985, Gillette et al, Biochemistry 33(10) 2927- 2937, 1994, and Jarman et al. Carcinogenesis 16(4), 683-688, 1993.

Pharmaceutically Acceptable Salts

[0075] The compounds of this invention can exist in free form for treatment, or where appropriate, as a pharmaceutically acceptable salt.

[0076] A "pharmaceutically acceptable salt" means any non-toxic salt of a compound of this invention that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this invention or an inhibitorily active metabolite or residue thereof. As used herein, the term "inhibitorily active metabolite or residue thereof" means that a metabolite or residue thereof is also an inhibitor of the PAR-2 signaling pathway. [0077] Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describe pharmaceutically acceptable salts in detail \r\ J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. These salts can be prepared in situ during the final isolation and purification of the compounds. Acid addition salts can be prepared by 1) reacting the purified free compound in its free-base form with a suitable organic or inorganic acid and 2) isolating the salt thus formed.

[0078] Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, glycolate, gluconate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2- hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like.

[0079] Base addition salts can be prepared by 1) reacting the purified free compound in its free acid form with a suitable organic or inorganic base and 2) isolating the salt thus formed. Salts derived from appropriate bases include alkali metal (e.g., sodium, lithium, and potassium), alkaline earth metal (e.g., magnesium and calcium), ammonium and N⁺(C₁.₄alkyl)₄ salts. This invention also envisions the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Water or oil-soluble or dispersible products may be obtained by such quaternization.

[0080] Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate. Other acids and bases, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid or base addition salts.

Pharmaceutically Acceptable Derivatives or Prodrugs

[0081] In addition to the compounds of this invention, pharmaceutically acceptable derivatives or prodrugs of the compounds of this invention may also be employed in compositions to treat or prevent the diseases, conditions and disorders. Specific examples are described below.

[0082] The compounds of this invention can also exist as pharmaceutically acceptable derivatives. A "pharmaceutically acceptable derivative" is an adduct or derivative which, upon administration to a patient in need, is capable of providing, directly or indirectly, a compound as otherwise described herein, or a metabolite or residue thereof. Examples of pharmaceutically acceptable derivatives include esters and salts of such esters.

[0083] A "pharmaceutically acceptable derivative or prodrug" means any pharmaceutically acceptable ester, salt of an ester or other derivative or salt thereof of a compound, of this invention which, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this invention or an inhibitorily active metabolite or residue thereof. Particularly favored derivatives or prodrugs are those that increase the bioavailability of the compounds of this invention when such compounds are administered to a patient (e.g., by allowing an orally administered compound to be more readily absorbed into the blood) or which enhance delivery of the parent compound to a biological compartment (e.g., the brain or lymphatic system) relative to the parent species.

[0084] Pharmaceutically acceptable prodrugs of the compounds of this invention include esters, amino acid esters, phosphate esters, metal salts and sulfonate esters.

Pharmaceutical Compositions

[0085] The present invention also provides chemical entities and compositions that are useful as inhibitors of the PA -2 signaling pathway.

[0086] In some aspects the present invention provides pharmaceutically acceptable compositions that comprise any of the chemical entities as described herein, and additionally comprise a pharmaceutically acceptable carrier, adjuvant or excipient.

[0087] The pharmaceutically acceptable carrier, adjuvant, or excipient, as used herein, includes any and all solvents, diluents, or other liquid vehicle, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like, as suited to the particular dosage form desired. Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1980) discloses various carriers used in formulating pharmaceutically acceptable compositions and known techniques for the preparation thereof. Except insofar as any conventional carrier medium is incompatible with the compounds of the invention, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutically acceptable composition, its use is contemplated to be within the scope of this invention.

[0088] Some examples of materials which can serve as pharmaceutically acceptable carriers include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, or potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, wool fat, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols; such a propylene glycol or polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen- free water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator.

[0089] The chemical entities of the invention can be formulated into pharmaceutical compositions for administration to animals or humans. In some embodiments, these pharmaceutical compositions comprise an amount of the PAR-2 signaling pathway inhibitor effective to treat or prevent the diseases or conditions described herein and a pharmaceutically acceptable carrier, adjuvant, or excipient. [0090] The exact amount of compound required for treatment will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the disease, disorder or condition, the particular agent, its mode of administration, and the like. The chemical entities of the invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The expression "dosage unit form" as used herein refers to a physically discrete unit of agent appropriate for the patient to be treated. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific effective dose level for any particular patient or organism will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed, and like factors well known in the medical arts.

[0091] In some embodiments, these compositions optionally further comprise one or more additional therapeutic agents. Some embodiments provide a simultaneous, separate or sequential use of a combined preparation.

Uses and Methods of Treatment

[0092] In some aspects, the present invention provides chemical entities that are inhibitors of the PA -2 signaling pathway and compositions comprising such chemical entities , as described above. In some aspects, the present invention provides methods and uses for treating or preventing a disease, condition, or disorder where PAR-2 is implicated in the disease, condition, or disorder, which employ administering a chemical entity of the invention, such as a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the invention comprising such chemical entity. Such methods and uses typically employ administering an effective amount of a chemical entity or pharmaceutical composition of the invention to a patient or subject.

[0093] The terms, "disease", "disorder", and "condition" may be used interchangeably herein to refer to any deviation from or interruption of the normal structure or function of any body part, organ, or system that is manifested by a characteristic set of symptoms and signs. Diseases, disorders and conditions of particular interest in the context of the present invention are those mediated by PA -2 or modulated the PAR-2 signaling pathway, or where PAR-2 is implicated in the disease state.

[0094] As used herein, the terms "subject" and "patient" are used interchangeably. The terms "subject" and "patient" refer to an animal (e.g., a bird such as a chicken, quail or turkey, or a mammal), particularly a mammal including non-primates (e.g., a cow, pig, horse, sheep, rabbit, guinea pig, rat, cat, dog, or mouse) and primates (e.g., a monkey, chimpanzee or human), and more particularly a human. In some embodiments, the subject is a non-human animal such as a farm animal (e.g., a horse, cow, pig or sheep), or a pet (e.g., a dog, cat, guinea pig or rabbit). In preferred embodiments, the subject is a human.

[0095] As used herein, an "effective amount" refers to an amount sufficient to elicit the desired biological response. In the present invention, certain examples of the desired biological reponse is to treat or preventa disease, condition, or disorder where PAR-2 is implicated in the disease state, to treat or prevent a PAR-2 mediated disease, condition, or disorder, to modulate the PAR- 2 signaling pathway, to inhibit the PAR-2 signaling pathway, or to enhance or improve the prophylactic or therapeutic effect(s) of another therapy used against a PAR-2-implicated or -mediated disease, condition, or disorder, or a disease, condition, or disorder modulated by the PAR-2 signaling pathway. The precise amount of compound administered to a subject will depend on the mode of administration, the type and severity of the disease, condition, or disorder and on the characteristics of the patient, such as general health, age, sex, body weight and tolerance to drugs. Persons skilled in the art will be able to determine appropriate dosages depending on these and other factors. When co-administered with other agents, an "effective amount" of the second agent will depend on the type of drug used. Suitable dosages are known for approved agents and can be adjusted by the person skilled in the art according to the condition of the patient, the type of condition(s) being treated and the amount of a compound described herein being used. For example, chemical entities described herein can be administered to a subject in a dosage range from between approximately 0.01 to 100 mg/kg body weight/day for therapeutic or prophylactic treatment. The chemical entities and compositions, according to the methods of the present invention, may be administered using any amount and any route of administration effective for eliciting the desired biological response.

[0096] As used herein, the terms "treat," "treatment" and "treating" can refer to both therapeutic and prophylactic treatments. For example, therapeutic treatments include the reduction or amelioration of the progression, severity and/or duration of one or more conditions, diseases or disorders and/or of one or more symptoms (specifically, one or more discernible symptoms) thereof, resulting from the administration of one or more therapies (e.g., one or more therapeutic agents such as a chemical entity or composition of the invention). In some embodiments, treatment refers to reduction or amelioration of the progression, severity and/or duration of one or more conditions, diseases or disorders, resulting from the administration of one or more therapies. In some embodiments, treatment refers to reduction or amelioration of the severity and/or duration of one or more conditions, diseases or disorders, resulting from the administration of one or more therapies. In some embodiments, treatment refers to reduction or amelioration of the progression, severity and/or duration of one or more symptoms (specifically, one or more discernible symptoms) of one or more conditions, diseases or disorders, resulting from the administration of one or more therapies. In some embodiments, treatment refers to reduction or amelioration of the severity and/or duration of one or more symptoms (specifically, one or more discernible symptoms) of one or more conditions, diseases or disorders, resulting from the administration of one or more therapies.

[0097] In some embodiments, the invention provides a method of treating a PA -2 mediated disease, condition, or disorder in a subject in need thereof.

[0098] In some embodiments, the invention provides a method for treating a disease, condition, or disorder where PAR-2 is implicated in the disease state.

[0099] In some embodiments, the invention provides a method for treating a disease, condition, or disorder where inhibition of PAR-2 signaling pathway is implicated in the treatment of the disease.

[0100] In some embodiments, the invention provides a method for treating a disease, condition, or disorder by modulating the PAR-2 signaling pathway.

[0101] In some embodiments, said disease, condition, or disorder is selected from such diseases, conditions, and disorders in which inhibitors of the PAR-2 signaling pathway may show therapeutic benefit. In some embodiments, said disease, condition, or disorder is selected from inflammatory disease or nociception (pain). In some embodiments, the nociception is caused by inflammation, cancer or injury. In some embodiments, said disease, condition, or disorder is selected from inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis), irritable bowel syndrome, asthma, rheumatoid arthritis, osteoarthritis, fibrosis (liver fibrosis, pulmonary fibrosis, cystic fibrosis, renal fibrosis, peritoneal fibrosis, pancreatic fibrosis, scleroderma, cardiac fibrosis, skin fibrosis, or intestinal fibrosis), gingivitis, periodontitis, vasculitis (e.g., Wegener's granulomatosis), atopic dermatitis, psoriasis, Netherton syndrome, systemic lupus erythematosus (SLE), scleroderma, interstitial lung disease, polymyositis, dermatomyositis, uveitis, Alzheimer's disease, Parkinson's disease, multiple sclerosis, inflammatory pain, postoperative incision pain, neuropathic pain, fracture pain, osteoporotic fracture pain, and gout joint pain in a patient. Additional diseases that show an increased proteolytic activity may benefit from inhibitors of the PA -2 signaling pathway.

[0102] In some embodiments, said disease, condition, or disorder is selected from diet-induced obesity, adipose inflammation, and metabolic dysfunction. In some embodiments, the metabolic dysfunction correlates with PAR-2 expression.

[0103] In some embodiments, said disease, condition, or disorder is selected from cancers. The term "cancer" means a disease characterized by unregulated cell growth. Examples of cancer for which the compounds of the invention can be used include colorectal cancer, pancreatic cancer, breast cancer, gastric cancer, ovarian cancer, squamous cell carcinoma, uterine endometrial cancer, nasopharyngeal carcinoma, esophageal adenocarcinoma, renal cell carcinoma and glioblastoma. Additional cancers that show an increased proteolytic activity or involvement of tissue factor and the coagulation cascade may benefit from inhibitors of the PAR-2 signaling pathway.

[0104] In some embodiments, said disease, condition, or disorder is selected from defects of excessive angiogenesis as manifested in solid tumor growth, tumor metastasis, multiple myeloma, lymphoma, ocular angiogenesis-mediated disorders (diabetic retinopathy, macular degeneration, and other ocular angiogenesis disorders), and angiogenesis-mediated inflammatory disorders.

[0105] In some embodiments, said disease, condition, or disorder is fibrosis. Examples of fibrosis include liver fibrosis, pulmonary fibrosis, cystic fibrosis, renal fibrosis, peritoneal fibrosis, pancreatic fibrosis, scleroderma, and cardiac fibrosis.

[0106] In some embodiments, the invention provides a method for treating or preventing inflammation, nociception (pain) or pruritus in a patient. In some embodiments the invention provides a method for treating inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis), irritable bowel syndrome, asthma, rheumatoid arthritis, osteoarthritis, fibrosis (liver fibrosis, pulmonary fibrosis, cystic fibrosis, renal fibrosis, peritoneal fibrosis, pancreatic fibrosis, scleroderma, cardiac fibrosis, skin fibrosis, or intestinal fibrosis), gingivitis, periodontitis, vasculitis (e.g., Wegener's granulomatosis), atopic dermatitis, psoriasis, Netherton syndrome, systemic lupus erythematosus (SLE), scleroderma, interstitial lung disease, polymyositis, dermatomyositis, uveitis, Alzheimer's disease, Parkinson's disease, multiple sclerosis, inflammatory pain, post-operative incision pain, neuropathic pain, fracture pain, osteoporotic fracture pain, and gout joint pain in a patient. Yet another embodiment provides a method for treating inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis) and osteoarthritis.

[0107] In some embodiments, the invention provides a method for treating or preventing inflammation or pain. In some embodiments, the invention provides a method for reducing inflammation.

[0108] In some embodiments of this invention provides a method for treating or preventing an inflammatory disease.

[0109] In some embodiments, the invention relates to a method of inhibiting the PAR-2 signaling pathway in a patient. In some embodiments, the invention provides a method for inhibiting PAR- 2 in a patient.

[0110] In some embodiments, the invention provides a method for inhibiting proteolytic activation of PAR-2 in a cell. In some embodiments, the invention provides a to a method of inhibiting PAR- 2 signaling pathway activity in a cell.

[0111] The invention also provides uses of a chemical entity or composition of the invention for the methods described above. In some embodiments, the invention provides uses in the manufacture of a medicament for such uses, for example, for treating a PAR-2 mediated disease in a patient, for treating or preventing inflammation or nociception (pain) in a patient, and for treating inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis), irritable bowel syndrome, asthma, rheumatoid arthritis, osteoarthritis, fibrosis (liver fibrosis, pulmonary fibrosis, cystic fibrosis, renal fibrosis, peritoneal fibrosis, pancreatic fibrosis, scleroderma, cardiac fibrosis, skin fibrosis, or intestinal fibrosis), gingivitis, periodontitis, vasculitis (e.g., Wegener's granulomatosis), atopic dermatitis, psoriasis, Netherton syndrome, systemic lupus erythematosus (SLE), scleroderma, interstitial lung disease, polymyositis, dermatomyositis, uveitis, Alzheimer's disease, Parkinson's disease, multiple sclerosis, inflammatory pain, postoperative incision pain, neuropathic pain, fracture pain, osteoporotic fracture pain, gout joint pain, fibrosis, cancer, diet-induced obesity, adipose inflammation, or metabolic dysfunction correlating with PAR-2 expression in a patient. In some embodiments, the invention provides uses in the manufacture of a medicament for treating inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis) or osteoarthritis. In some embodiments, the invention provides uses in the manufacture of a medicament for use in treating inflammation or pain.

[0112] In some aspects, the invention provides the use of a chemical entity or composition of the invention in the manufacture of a medicament for use in inhibiting proteolytic activation of PA - 2 in a cell. In some aspects, the invention provides a chemical entity or composition of the invention in the manufacture of a medicament for inhibiting PAR-2 activity in a cell.

[0113] In some embodiments, the invention provides co-administering to a patient an additional therapeutic agent, wherein said additional therapeutic agent is appropriate for the disease, condition or disorder being treated; and said additional therapeutic agent is administered together with a chemical entity of the invention as a single dosage form, or separately from said compound as part of a multiple dosage form.

[0114] As used herein, the terms "in combination" or "co-administration" can be used interchangeably to refer to the use of more than one therapy (e.g., one or more prophylactic and/or therapeutic agents). The use of the terms does not restrict the order in which therapies (e.g., prophylactic and/or therapeutic agents) are administered to a patient, nor does it require administration in any specific proximity in time, so long as in the judgment of a suitable physician the patient is understood to be receiving the one or more therapies at the same time. For example, receiving therapy A on days 1-5 of a 28-day schedule and therapy B on days 1, 8 and 15 of a 21-day schedule would be considered "in combination" or a "co-administration".

[0115] Co-administration also encompasses administration of the first and second amounts of the compounds of the co-administration in an essentially simultaneous manner, such as in a single pharmaceutical composition, for example, capsule or tablet having a fixed ratio of first and second amounts, or in multiple, separate capsules or tablets for each. In addition, such coadministration also encompasses use of each compound in a sequential manner in either order.

Modes of Administration and Dosage Forms

[0116] The pharmaceutically acceptable compositions of this invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (as by powders, ointments, or drops), bucally, as an oral or nasal spray or via inhalation, or the like, depending on the identity and/or severity of the disease being treated. In certain embodiments, the chemical entities of the invention may be administered orally or parenterally at dosage levels of about 0.01 mg/kg to about 50 mg/kg, and preferably from about 1 mg/kg to about 25 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect.

[0117] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0118] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S. P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

[0119] The injectable formulations can be sterilized, for example, by filtration through a bacterial- retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0120] In order to prolong the effect of a compound of the present invention, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends upon its rate of dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide- polyglycolide. Depending upon the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.

[0121] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

[0122] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

[0123] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.

[0124] The active compounds can also be in microencapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

[0125] Dosage forms for topical or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, eardrops, and eye drops are also contemplated as being within the scope of this invention. Additionally, the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0126] The compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intraarticular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally or intravenously. [0127] Sterile injectable forms of the compositions of this invention may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally- acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or di-glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents which are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants, such as Tweens, Spans and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.

[0128] The pharmaceutical compositions of this invention may be orally administered in any orally acceptable dosage form including capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavouring or colouring agents may also be added.

[0129] Alternatively, the pharmaceutical compositions of this invention may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

[0130] The pharmaceutical compositions of this invention may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs. [0131] Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches may also be used.

[0132] For topical applications, the pharmaceutical compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of this invention include mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

[0133] For ophthalmic use, the pharmaceutical compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic uses, the pharmaceutical compositions may be formulated in an ointment such as petrolatum.

[0134] The pharmaceutical compositions of this invention may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.

[0135] The amount of inhibitor that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated, the particular mode of administration. Preferably, the compositions should be formulated so that a dosage of between 0.01 - 100 mg/kg body weight/day of the inhibitor can be administered to a patient receiving these compositions.

[0136] It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated. The amount of inhibitor will also depend upon the particular compound in the composition.

Administering with Another Agent

[0137] Depending upon the particular conditions to be treated or prevented, additional drugs, which are normally administered to treat or prevent that condition, may be administered together with the chemical entities of this invention.

[0138] Those additional agents may be administered separately, as part of a multiple dosage regimen. Alternatively, those agents may be part of a single dosage form, mixed together with the inhibitor of the PAR-2 signaling pathway in a single composition.

Biological Samples

[0139] As inhibitors of the PAR-2 signaling pathway, the chemical entities and compositions of this invention are also useful in biological samples. In some aspects, the invention relates to inhibiting PAR-2 activity in a biological sample, which method comprises contacting said biological sample with a chemical entity described herein or a composition comprising said chemical entity. The term "biological sample", as used herein, means an in vitro or an ex vivo sample, including cell cultures or extracts thereof; biopsied material obtained from a mammal or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof. The term "chemical entities described herein" includes chemical entities of formula I.

[0140] Inhibition of PAR-2 signaling pathway activity in a biological sample is useful for a variety of purposes that are known to one of skill in the art. Examples of such purposes include blood transfusion, organ-transplantation, and biological specimen storage.

Study of GPCRs

[0141] In some aspects, the invention relates to the study of GPCRs in biological and pathological phenomena; the study of pathways mediated by such GPCRs; and the comparative evaluation of new GPCRs. Examples of such uses include biological assays such as enzyme assays and cell- based assays.

[0142] The activity of the compounds as inhibitors of the PAR-2 signaling pathway may be assayed in vitro, in vivo or in a cell line. In vitro assays include assays that determine inhibition of either synthetic activators of PAR-2 such as SLIGKV-NH2 or protease-dependent activators such as trypsin activation of PAR-2. [0143] In some aspects, the invention provides a method for modulating PAR-2 activation by contacting a compound described herein with PAR-2.

Synthetic Methods

[0144] In general, the chemical entities of the invention can be prepared by methods described herein or by other methods known to those skilled in the art. Exemplary preparations of the chemical entities of the invention are described below.

[0145] The chemical entities of the disclosure may be prepared in light of the specification according to the schemes below as well as according to steps generally known to those of ordinary skill in the art. The chemical entities may be analyzed by known methods such as LC-MS (liquid chromatography/mass spectrometry) and NMR (nuclear magnetic resonance).

[0146] The phrase "standard aqueous workup" refers to removal of water miscible solvents (if applicable) and partitioning the reaction mixture between a suitable organic solvent (chosen from EtOAc, DCM, hexanes, Et₂0, heptane, MTBE, or 2-methyl-THF), followed by separation of the layers, extracting the aqueous phase 1-6 times more, combination of the organic phases and washing with H₂0, saturated aqueous NaCI (brine), drying over Na₂S0₄ or MgS0₄, followed by filtration and removal of the solvent in vacuo.

[0147] Mass spectrometry samples were analyzed on a Waters UPLC Acquity mass spectrometer operated in single MS mode with electrospray ionization. Samples were introduced into the mass spectrometer using chromatography. Mobile phase for the mass spectrometry analyses consisted of 0.1% formic acid and acetonitrile-water mixture. As used herein, the term "Rt time" refers to the LC-MS retention time, in minutes, associated with the chemical entity. Unless otherwise indicated, the LC-MS methods utilized to obtain the reported retention time are as detailed below:

Method A: 5%-85% acetonitrile-water over 6 minutes run time, Waters AcquityHSS T3 1.8μιη, 2.1 mm ID x50 mm. Flow rate is 1.0 mL/min.

Method B: 50%-100% acetonitrile-water over 6 minutes run time, Waters AcquityHSS T3 1.8μιη, 2.1 mm ID x50 mm. Flow rate is 1.0 mL/min.

Method C: 5%-85% acetonitrile-water over 3 minutes run time, Waters AcquityHSS T3 Ι.δμιη, 2.1 mm ID x50 mm. Flow rate is 1.0 mL/min. Method D: 10%-100% acetonitrile-water over 6 minutes run time, Waters XSelect CHS C18 2.5μιη, 4.6 mm ID x30 mm Column XP. Flow rate is 1.0 mL/min.

Method E: 5%-85% acetonitrile-water over 6 minutes run time, Waters Acquity UPLC^® HSS C18 SB Ι.δμιη, 2.1 mm ID x50 mm. Flow rate is 1.0 mL/min.

Method F: 5-85% acetonitrile-water over 2 minutes run time, Waters AcquityHSS T3 Ι.δμιη, 2.1 mm ID x50 mm. Flow rate is 1.0 mL/min

Method G: 50%-90% acetonitrile-water over 6 minutes run time, Waters AcquityHSS T3 Ι.δμιη, 2.1 mm ID x50 mm. Flow rate is 1.0 mL/min.

Method H: 10%-100% acetonitrile-water over 5 minutes run time, Waters XSelect CHS™ C18 2.5μιη, 4.6 mm ID x30 mm Column XP. Flow rate is 1.0 mL/min.

Method I: 50-100% acetonitrile-water over 2 minutes run time, Waters AcquityHSS T3 Ι.δμιη, 2.1 mm ID x50 mm. Flow rate is 1.0 mL/min.

Method J: 15%-98% acetonitrile-water over 1.5 minutes run time, Waters AcquityHSS T3

Ι.δμιη, 2.1 mm ID x30 mm. Flow rate is 1.5 mL/min.

Method K: 0%-50% acetonitrile-water over 6 minutes run time, Waters AcquityHSS T3 Ι.δμιη, 2.1 mm ID x50 mm. Flow rate is 1.0 mL/min.

Method L: 0%-50% acetonitrile-water over 3 minutes run time, Waters AcquityHSS T3 Ι.δμιη, 2.1 mm ID x50 mm. Flow rate is 1.0 mL/min.

Method M : 15-9δ% acetonitrile-lOmM ammonium bicarbonate, pH 10 in water over 1.5 minutes run time, Waters CSHC18, 1.7um, 2.1 mm ID x 30 mm. Flow rate is 1.3 mL/min. 8] Purification by reverse phase HPLC is carried out under standard conditions using a Phenomenex Gemini 21.2 mm ID x 250 mm column, 5 μ and Gemini 21.2 mm ID x 75 mm column, 5 μ, 110A. Elution is performed using a linear gradient CH₃CN-H₂0 (with or without 0.01% TFA or formic acid buffer) as mobile phase. Solvent system is tailored according to the polarity of the compound, Flow rate, 20 mL/min. Compounds are collected either by UV or Waters 3100 Mass Detector, ESI Positive Mode. UV directed reverse phase purification is also carried out on a Isco Combiflash EZ PREP equipped with a Gemini-NX 30X250 mm, C18, 5 micron column, using mixtures of MeCN-H₂0 tailored to the polarity of the compound with 0.1% formic acid or 10 mM Ammonium bicarbonate as modifier. [0149] Fractions containing the desired compound are combined, concentrated (rotary evaporator) to remove excess CH₃CN and the resulting aqueous solution is lyophilized to afford the desired material.

[0150] Nuclear magnetic resonance (NMR) spectra are recorded on INOVA 400 MHz Varian or a Bruker Ascend™ 400 MHz instrument. The residual solvent protons (^XH) are used as internal standards. The following solvents are used: chloroform-d (CDCI₃), methanol-d₄ (CD₃OD), DMSO- d₆. ^XH NM R data are presented as follows: chemical shift in ppm downfield from tetramethylsilane (multiplicity, coupling constant, integration). The following abbreviations are used in reporting NMR data: s, singlet; d, doublet; t, triplet; q, quartet; p, pentuplet; h, hextuplet; dd, doublet of doublets; ddd, doublet of doublets of doublets; dddd, doublet of doublets of doublets of doublets; dt, doublet of triplets; dtd, doublet of triplets of doublets; ddt, doublet of doublets of triplets; dq, doublet of quartets; dp, doublet of pentuplets; td, triplet of doublets; qd, quintet of doublets; m, multiplet.

[0151] Purification by flash chromatography on silica gel is carried out under standard conditions using, but not restricted to, either of the following instruments and supplies: Biotage^® SP1 or SP2 purification system with Biotage^® SNAP Cartridge KP-Sil column lOg, 25g, 50g, lOOg or 340g or, Biotage^® SNAP Cartridge KP-C18-HS 12g, 30g, 60g or 120g and, CombiFlash^®Rf Teledyne Isco purification system with Silica RediSep^®Rf normal phase column 12g, 24g, 40g, 80g, 120g, 220g or 330g. Solvent system is tailored according to the polarity of the compound. Fractions containing the desired compound are combined and concentrated (rotary evaporator) to remove the solvent and to afford the desired material.

List of Abbreviations

[0152] The following abbreviations are used in the examples below:

Ac acetyl

AcOH acetic acid

ACN acetonitrile

Ac₂0 acetic anhydride

aq aqueous

BF₃-OEt₂ boron trifluoride diethyl ether

Bn benzyl CDI carbonyl diimidazole

CH3CN acetonitrile

Cone. concentrate

CV column volume

D ²H (deuterium)

DBU l,8-diazabicyclo[5.4.0]undec-7-ene

DCM dichloromethane (or methylene chloride)

DIPEA A/,/\/-diisopropylethylamine

DMAP 4-dimethylaminopyridine

DM F dimethylformamide

DMSO dimethylsulfoxide

dppf diphenylphosphinoferrocene

Equiv equivalent

EtOAc ethyl acetate

FA formic acid

g gram(s)

HATU O-iy-azabenzotriazol-l-yl^/V^/V" Λ -tetramethyluroniumhexafluorophosphate h hour(s)

Hex hexanes

HPLC high performance liquid chromatography

JohnPhos (2-biphenyl)-di-tert-butylphosphine

LCMS liquid chromatography mass spectrometry

M molar

MHz megahertz

mg milligram(s)

mL milliliter(s)

mM millimolar

MeOH methanol

MeONa sodium methoxide min minute(s)

MS mass spectrometry

MTBE methyl ieri-butyl ether

μΜ micromolar

N normal (molar) concentration

¹H NM proton nuclear magnetic resonance

NMO /V-methylmorpholine-ZV-oxide

ON overnight

Pd₂(dba)₃ tris(dibenzylideneacetone)dipalladium(0)

Pd/C palladium on carbon

Pd(OAc)₂ palladium(ll) acetate

PdCI₂(dppf).DCM (l,l'-bis-(diphenylphosphino)-ferrocene)palladium (II) dichloride

Pd(PPh₃)₄ tetrakis(triphenylphospine)palladium (0)

psi pound per square inch

Py pyridine

r.b.f. (rbf) round bottom flask

RT (rt or r. t.) room temperature

RuPhos 2-dicyclohexylphosphino-2',6'-diisopropoxybiphenyl

S-Phos 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl

T3P propylphosphonic anhydride

TBAF tetrabutylammonium fluoride

TBDMSOTf ieri-butyldimethylsilyl trifluoromethanesulfonate

TBS ieri-butyldimethylsilyl

TEA triethylamine

Tf trifluoromethanesulfonyl

TFA trifluoroacetic acid

TFAA trifluoroacetic anhydride

THF tetrahydrofuran

TLC thin layer chromatography TMS trimethylsilyl

TMSI trimethylsilyl iodide

TMSN3 trimethylsilyl azide

TMSOTf trimethylsilyl trifluoromethanesulfonate

TPAP tetrapropylammonium perruthenate

UPLC ultra performance liquid chromatography

XPhos 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl

XantPhos 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene

[0153] The following generic schemes and examples illustrate how to prepare the compounds of the present disclosure.

General Synthetic Route 1

General Synthetic Route 2

R = H, Me

General Synthetic Route 4

General Synthetic Route 6

General Synthetic Route 8

General Procedure 1: Amide Formation

[0154] The appropriate carboxylic acid (1.0 equiv) is dissolved in DM F or NMP (0.03 to 0.4M) before HATU (1.1 to 1.5 equiv), or T3P (1 to 5 equiv) the corresponding amine (1.0 to 1.5 equiv) and Hunig's base (3.0 to 5.0 equiv) are added (the addition order of the reagents may vary) The mixture is stirred at room temperature for 45 min. to 72h. Any one of these 3 work-up procedures can be employed:

1. Water or aq. ammonium chloride is added and the aq. phase is extracted with EtOAc. The combined organic phase is washed with brine, dried over MgS0₄, filtered and the filtrate evaporated under reduced pressure affording the title compound which is used in the subsequent step without further purification.

2. Water or aq. ammonium chloride is added and the aq. phase is extracted with EtOAc. The combined organic phase is washed with IN HCI, sat. aq. NaHC0₃ and brine, dried over Na₂S0₄, filtered and the filtrate evaporated under reduced pressure affording the title compound which is used in the subsequent step without further purification.

3. The volatiles are removed under reduced pressure and the residue is purified by flash chromatography on silica gel or by mass-directed reverse phase HPLC, affording the title compound.

General Procedure 2: tert-Butyl Carbamate Deprotection

[0155] The appropriate boc-protected amine (1.0 equiv) is dissolved in an appropriate solvent such as DCM, 1,4-dioxane or MeOH (or combinations thereof) (0.1 M) before an acid such as 4N HCI solution in 1,4-dioxane or TFA (20.0 to 30.0 equiv) are added. The solution is stirred at room temperature for 1 to 4h before the volatiles are removed under reduced pressure, affording the title compound which is used in the subsequent step without further purification. In some cases, MeOH is added to solubilize the salts and the resulting solution is allowed to stir for an additional lh prior to concentration under reduced pressure.

General Procedure 3: Amide Formation

[0156] A solution of the appropriate piperazine (1.0 equiv) and HATU (1.3 equiv) in NMP or DMF (400 μί to 800 μί) is added to either the corresponding NMP solution of carboxylic acid (200 μί, 0.1 mM), or the corresponding neat acid (1.0 to 1.1 equiv). Hunig's base (3.0 to 5.0 equiv) is added and the solution stirred at room temperature for 20 minutes to 16 hours. Any one of these 3 work-up procedures can be employed: 1. Aq. ammonium chloride is added and the aq. phase is extracted with EtOAc. The combined organic phase is washed with brine, dried over MgS0₄, filtered and the filtrate evaporated under reduced pressure affording the title compound which is used in the subsequent step without further purification.

2. Aq. ammonium chloride is added and the aq. phase is extracted with EtOAc. The combined organic phase is washed with IN HCI, sat. aq. NaHC0₃ and brine, dried over Na₂S0₄, filtered and the filtrate evaporated under reduced pressure affording the title compound which is used in the subsequent step without further purification.

General Procedure 4: Pyridine Synthesis

[0157] To a solution of the appropriate acetophenone (1.05 to 1.3 equiv), aldehyde (1.0 equiv) and ethyl 2-cyanoacetate (1.0 equiv) in ethanol (1.0 M) is added ammonium acetate (7.0 to 10 equiv) and the solution is stirred at 80°C for 6 to 18h. The solution is cooled to room temperature before water is added and the resulting precipitate recovered by filtration and dried under reduced pressure. If only partial aromatization is observed, the solid is dissolved in DCM (0.25 to 0.9 M) and DDQ (0.5 to 1.0 equiv) is added. The mixture is stirred at room temperature for 4 to 96h, filtered and the filtrate evaporated under reduced pressure. The residue obtained is triturated from hot ethanol and water, or methanol at room temperature affording the title product as a solid.

General Procedure 5: lodination

[0158] To a stirring suspension of the appropriate 2-pyridone (1 equiv) and pyridine (1.1 to 1.3 equiv) in acetonitrile (0.5 to 1 M) at 0°C is added dropwise triflic anhydride (1.05 to 1.25 equiv) and the resulting solution is stirred at room temperature for 30 minutes. Nal (2 to 10 equiv) and HCI 12N (1.1 to 5 equiv) or triflic acid (1 to 1.25 equiv) are added and the solution stirred at 90°C for 16h. Water and aq. NaHC0₃ is added and the aq. phase is extracted with EtOAc or DCM. The combined organic phase is washed with brine, dried over MgS0₄, filtered, and the filtrate is evaporated under reduced pressure. The residue is purified either by flash chromatography on silica gel, or by trituration affording the title compound as a solid.

General Procedure 6: Alternative lodination Conditions [0159] To a stirring suspension of the appropriate 2-pyridone (1 equiv) and pyridine (1.1 to 1.3 equiv) in acetonitrile (0.5 to 1 M) at 0°C is added dropwise triflic anhydride (1.05 to 1.25 equiv) and the resulting solution is stirred at room temperature for 30 minutes. The volatiles are removed under reduced pressure and the residue dissolved in N MP (0.5 to 1M) before Nal (2 to 10 equiv) and HCI 12N (1.5 to 5 equiv), or triflic acid (1.5 to 5 equiv) are added and the solution stirred at 80°C or 90°C for 16h. Water and aq. NaHC0₃ are added, and the aq. phase is extracted with EtOAc. The combined organic phase is washed with brine, dried over MgS0₄, filtered and the filtrate evaporated under reduced pressure. The residue is purified either flash chromatography on silica gel or trituration affording the title compound as a solid.

General Procedure 7: Nitrile Hydrolysis

[0160] To a round bottom flask containing the appropriate 2-iodo-pyridine-3-carbonitrile (1 equiv) is added sulfuric acid (10 equiv) and the mixture is stirred at 80°C to 90°C for 4 to 72h. The solution is cooled to room temperature and added dropwise to a saturated solution of sodium carbonate or potassium carbonate in water (10 to 20 equiv). The resulting white precipitate is collected by filtration, washed with water and dried under reduced pressure, affording the product as a white solid.

General Procedure 8: Hofmann Rearrangement

[0161] To a solution of KOH (7 equiv) in water (2.6 volume equiv) at 0°C is added bromine (1.3 equiv) dropwise and the solution stirred for 5 minutes before the appropriate 2-iodo-pyridine-3- carboxamide (1 equiv) is added portion wise. The suspension is stirred at 0°C for 5 minutes before careful addition of water (0.9 volume equiv) and THF (0.9 volume equiv) and stirring is pursued for 15 minutes. The solution is stirred at room temperature for 10 minutes to 2 hours. Water is added and the aq. phase is extracted with EtOAc. The combined organic phase is washed with sat. aqueous Na₂S0₃ and brine, and dried over MgS0₄ and filtered. The filtrate is evaporated under reduced pressure affording the title compound which is used in the subsequent step without further purification or purified by flash chromatography on silica gel.

General Procedure 9: Sandmeyer Reaction

[0162] To a solution of the appropriate 3-aminopyridine (1.0 equiv) in TFA (4.7 volume equiv) is added dropwise isopentyl nitrite (1.5 to 6.0 equiv) and the solution stirred for 30 minutes. The solution is diluted with methanol (10 to 20 volume equiv) and K₂C0₃ is added until pH 12 is obtained. The solution is stirred for 1 to 16h before water is added, the pH is adjusted to 5 using 12N HCI and the aq. phase is extracted with EtOAc or DCM. The combined organic phase is washed with brine, dried over MgS0₄ and filtered. The filtrate is evaporated under reduced pressure affording the title product compound which is used in the subsequent step without further purification or purified by flash chromatography on silica gel.

General Procedure 10: Sonogashira Cross-Coupling

[0163] To a round bottom flask containing a solution of the appropriate 2-iodo-3-hydroxy-pyridine (1.0 equiv) in THF (5.1 volume equiv) is added potassium carbonate (7.0 equiv), PdCI₂(PPh₃)₂ (0.04 equiv) and Cul (0.08 equiv) and the solution is warmed to 80°C. A solution of ethyl prop-2- ynoate (2.5 to 4.0 equiv) in THF (2.6 volume equiv) is added dropwise over 30 minutes to 1 hour. The solution is cooled to room temperature and acidified using 12N HCI. Water is added and the aq. phase is extracted with EtOAc. The combined organic phase is washed with brine, dried over MgS0₄, filtered and the filtrate evaporated under reduced pressure. The residue is purified by flash chromatography on silica gel affording the title compound as a solid.

General Procedure 11: Ester Hydrolysis

[0164] Method A: To a solution of the appropriate ester (1.0 equiv) in solvents selected from ethanol, methanol, dioxane, THF, NMP, DMF and DMSO, or combinations thereof (0.15 to 0.3 M) is added a 2M aqueous solution of LiOH (2.0-5.0 equiv), NaOH (2.0-5.0 equiv), or KOH (2.0-5.0 equiv), and the solution is stirred for lh to 3 days at temperature ranging from r.t. to 80 °C. Any one of these 5 work-up procedures can be employed:

1. Water is added and the solution acidified to pH 2-4 using an appropriate acid such as 1 to 12N HCI, formic acid or aqueous citric acid. The precipitate is collected by filtration, washed with water and dried under reduced pressure, affording the title compound as a solid. Alternatively the compound can be dissolved in DMSO and purified by mass-directed reverse-phase preparative HPLC affording the desired compound.

2. Water is added, the solution acidified to to pH 2-4 using an appropriate acid such as 1 to 12N HCI or aqueous citric acid or formic acid and the aq. phase is extracted with EtOAc. The combined organic phase is washed with brine, dried over MgS0₄, filtered and the filtrate evaporated under reduced pressure affording the title compound as a solid. Alternatively the compound can be dissolved in DMSO or NMP and purified by mass-directed reverse- phase preparative HPLC affording the desired compound.

3. Water is added and the solution acidified using 12N HCI. The precipitate is collected by filtration, washed with water and dried under reduced pressure affording the title compound as a solid. Alternatively the compound can be dissolved in DMSO and purified by mass-directed reverse-phase preparative HPLC affording the desired compound.

4. The reaction mixture is filtered and purified by mass-directed reverse-phase preparative HPLC affording the desired compound.

5. The reaction mixture is treated with Amberlite I 120(H), filtered, concentrated and purified by mass-directed reverse-phase preparative HPLC affording the desired compound.

[0165] Method B. A solution of the ethyl ester (1.0 equiv) in t-BuOH (0.05 to 0.5 M) was heated at temperatures ranging from RT to 100°C and treated with KOtBu (2 to 8 equiv). The reaction is heated until judged complete by LCMS. The reaction is cooled to RT and poured slowly into aqueous HCI (excess). The precipitate is filted and the solid dried under vacuum. The product may also be isolated by standard aqueous workup. In some cases, it proves advantageous to treat the cooled reaction mixture with a stoichiometric amount (relative to KOtBu) of acetic acid before treatment with HCI.

General Procedure 12: Sulfonamide Formation

[0166] To a solution of the appropriate amine (1.0 equiv) in DMF (0.05-0.25M) is added triethylamine (2.5 to 5.0 equiv) followed by the appropriate sulfonyl chloride (1.2 to 2.4 equiv). The resulting solution is stirred at a temperature ranging from room temperature to 50°C for 4 to 16 hours. Upon reaction completion, the crude mixture is purified directly by mass-directed reverse-phase HPLC affording the desired compound.

General Procedure 13: Ester Hydrolysis

[0167] To a solution of the appropriate piperazine amide (1.0 equiv) in dioxane (0.15 to 0.3 M) is added a 2M aqueous solution of LiOH or NaOH (2.0-4.0 equiv) and the solution is stirred for 1- 22h at rt or reflux. Either one of these 2 work-up procedures can be employed:

1. Water is added, the solution acidified to pH 4 using 12 N HCI and the aq. phase is extracted with EtOAc. The combined organic phase is washed with brine, dried over MgS0₄, filtered and the filtrate evaporated under reduced pressure affording the title compound as a solid.

2. The solution is acidified to pH 4 using 4N HCI in dioxane and evaporated in vacuo. The residue is dissolved in DMSO and purified by mass-directed reverse-phase preparative HPLC affording the desired compound. General Procedure 14: Alkylation or arylation of amides

[0168] To a solution of the appropriate piperazinone (1.0 equiv) in NMP (0.1-0.25M) is added 60% sodium hydride (1.2equiv). The resulting solution is stirred at a room temperature for 15-30 minutes before the appropriate haloalkyl or haloaryl/haloheteroaryl (1.0 to 5.0 equiv) is added. The mixture is stirred at a temperature from T to 150°C for 1 to 16 hours. Upon completion of the reaction, the crude mixture is purified by mass-directed reverse-phase affording the desired compound.

General Procedure 15: Suzuki cross-coupling

[0169] To a rbf containing a solution of the appropriate aryl halide (1 equiv) in dioxane (0.03 to 0.4M) is added Pd₂dba₃.CHCl3 (0.02 to 0.25 equiv) and S-Phos (0.05 to 0.5, 2 equiv of Pd atom). The resulting solution is degassed with nitrogen for 5 minutes prior to addition of respectively K3PO4 (2 to 5 equiv. of a 2M aq. solution) and associated boronic acid or boronate ester (1 to 2.5 equiv). It is then stirred between 60 and 105°C for 2 to 18h. DCM or EtOAc and NaHC0₃ are added to the mixture, the layers are separated and the aqueous phase is extracted with DCM or EtOAc twice. The combined organic layers are dried with MgS0₄, filtered and concentrated under reduced pressure to afford a crude product which is used in the subsequent step withouth further purification or purified by filtration over silica gel or mass-directed reverse-phase HPLC.

General Procedure 16: Hydrogenation

[0170] To a flask containing palladium on carbon or platinum oxide (0.05 to 0.2 equiv) and the appropriate olefin (1 equiv) in EtOH or MeOH (0.03 to 0.1 M) or MeOH:DCM (under a nitrogen atmosphere is evacuated under reduced pressure and re-filled with hydrogen 3 times and the suspension is stirred under hydrogen pressure at room temperature overnight. The suspension is filtered over celite, washed with DCM and the volatiles are removed under reduced pressure. The residue is purified by mass-directed reverse phase HPLC to afford upon lyophilization the desired compound.

General Procedure 17: Silyl ether protection

[0171] To a solution of the appropriate primary alcohol (1 equiv) and trialkyl chlorosilane (1.5 equiv) in DM F (0.6 M) is added imidazole (2.5 equiv) at rt and the solution is stirred at rt for 18h.

[0172] Aq. sat. NaHC0₃ is added and the aq. phase is extracted with EtOAc. The combined organic phase is washed with IN HCI, sat. aq. NaHC0₃ and brine, dried over Na₂S0₄, filtered and the filtrate evaporated under reduced pressure affording the title compound which is used in the subsequent step without further purification.

[0173] Aq. and EtOAc are added to the reaction, the organic phase is extracted with EtOAc to afford the title compound used as is for the next step without further purification.

General Procedure 18: C-7 Selective Addition

[0174] To a round bottom flask containing a solution of tert-butyl-[(5-chloro-4-oxido-furo[3,2- b]pyridin-4-ium-2-yl)methoxy]-diphenyl-silane (1 equiv) and iodocopper (0.4 equiv) in THF (0.5 M) at -5°C is added trifluoro-tetrahydrofuran-l-ium-l-yl-boron (2.2 equiv) and the solution is allowed to stir for 30 minutes. The mixture is kept at -5°C and to this is slowly added tert- butyl(chloro)magnesium (2.4 equiv) over 60 minutes. Aq. ammonium chloride is added and the aq. phase is extracted with DCM. The phases are seperated, DDQ (0.11 equiv) is directly added to the organic phase and the mixture is allowed to stir for 20 minutes at room temperature. DDQ (0.10 equiv) is added again and the mixture is allowed to stir for 4h at rt. Water is added, the phases are seperated and the volatiles are removed under reduced pressure affording the title compound which is used in the subsequent step without further purification

General Procedure 19: Silyl ether deprotection

[0175] To a round bottom flask containing a solution of tert-butyl-[(7-tert-butyl-5-chloro-furo[3,2- b]pyridin-2-yl)methoxy]-diphenyl-silane (1 equiv) in TH F (0.4 M ) at rt is added TBAF (2 equiv) and the solution is allowed to stir for 3h. Water along with DCM is added and the phases are seperated. The solvents are evaporated under reduced pressure and the residue purified by flash chromatography on silica gel affording the title compound as a solid.

General Procedure 20: Lev Oxidation

[0176] To a solution of the appropriate primary alcohol (1 equiv) and N MO monohydrate (10 equiv) in acetonitrile (0.15 to 0.2 M ) in a water bath is added TPAP (0.05 equiv to 0.1 equiv) portionwise and the solution is stirred at room temperature for lh. The solution is cooled to 0°C, /^'-PrOH (10 equiv) is added slowly and the reaction is stirred for 15 to 30 minutes. The reaction mixture is diluted with water and the volatiles are removed under reduced pressure. The aq. phase is basified to pH 13 using 2N NaOH and washed with MTBE and the organic phase discarded. The aq. phase is acidified to pH 3 with 2N HCI, the product formed is collected by filtration, washed with water and heptane and dried under reduced pressure affording the title compound as a solid. General Procedure 21: TFA deprotection of PMB protected amine

[0177] A solution of the appropriate poro-methoxybenzyl protected amine (1 equiv) in trifluoroacetic acid (0.06 to 0.2 M) is heated in the microwave or thermally at temperatures ranging from T to 130°C for the appropriate time to complete the reaction. The trifluoroacetic acid is removed under reduced pressure. The residue is purified by mass-directed reverse phase HPLC to afford upon lyophilization the desired compound.

General Procedure 22: C-N aryl bond formation

[0178] Method A: The appropriate piperazine (1.0 equiv) is dissolved in DM F, dioxane, DMSO or NM P (0.01 to 0.4M), treated with a base selected from Cs₂C0₃, DBU and Hunig's base (2.0 to 5.0 equiv) and the appropriate aryl halide (1.0 to 5.0 equiv). The mixture is stirred at a temperature ranging from r.t. to 150 °C for 30 min to 24h, in the microwave of thermally.

[0179] Method B: The appropriate piperazine or piperazinone (1.0 equiv) is dissolved in DM F, dioxane, DMSO or NMP (0.01 to 0.4M), treated with a base selected from NaOtBu, K₃P0₄ and K₂C0₃ (2.0 to 5.0 equiv), the appropriate aryl halide (1.0 to 5.0 equiv) and the solution is flushed with nitrogen prior to adding a palladium catalyst selected from palladium acetate, RuPhos(IV), and a ligand selected from RuPhos, JohnPhos, XPhos and XantPhos. The mixture is stirred at a temperature ranging from r.t. to 150 °C for 30 min to 24h, in the microwave of thermally.

[0180] Method C: The appropriate piperazinone (1.0 equiv) is dissolved in toluene, DMF, dioxane or NM P (0.01 to 0.4M), treated with a base selected from Cs₂C0₃, K₃P0₄ and K₂C0₃ (2.0 to 5.0 equiv), added the appropriate aryl halide (1.0 to 5.0 equiv) and Cul (1.0 equiv to 3 equiv), optionally with ligands such as Λ/,Λ/'-dimethylethylenediamine or Ν,Ν,Ν'Ν'- tetramethylethylenediamine (0.1 equiv to 3 equiv). The mixture is stirred at a temperature ranging from RT to 150 °C for 30 min to 24h, in the microwave of thermally.

[0181] Method D: Under nitrogen, sodium hydride 60% dispersed in mineral oil (1.15 eq) is added to a stirred mixture of the appropriate piperazinone (1.0 eq) in DMF. The mixture is stirred for 15 min-1 hour at rt and then, the appropriate aryl halide (1.5 eq) is added. The reaction mixture is stirred for 2-16 hours at 60°C thermally.

[0182] Any one of these 3 work-up procedures can be employed: 1. Water is added, and the aq. phase is extracted with EtOAc. The combined organic phase is washed with brine, dried over MgS0₄, filtered and the filtrate evaporated under reduced pressure affording the title compound as a solid.

2. The volatiles are removed under reduced pressure and the residue is purified by flash chromatography on silica gel or by mass-directed reverse phase HPLC, affording the title compound.

3. Water is added (a solid crashed out) and the resulting precipitate is allowed to stir for lh.

The heterogenous mixture is filtered on Buchner and the desired solid is washed with water, followed by heptane to give the desired product.

[0183] In the case where the product is substituted with an ester functional group, the ester group may be hydrolyzed by either of General Procedure 11 or General Procedure 23.

General Procedure 23: Acidic ester hydrolysis

[0184] A solution of the appropriate tert-butylester (1.0 equiv) in solvents selected from methanol, ethanol, dioxane, THF, NMP, DCM or DMF (or combinations thereof) (0.01 to 1 M) is treated with an appropriate acid such as HCI in dioxane (1.0 equiv to 10 equiv) or aqueous HCI (1.0 equiv to 10 equiv) or TFA and stirred at a temperature ranging from r.t. to 80°C for 30 min to 24h.

General Procedure 24: Preparation of Aryl tetrazoles

Step I:

[0185] Reaction is carried out using the appropriate heteroaryl nitrile and 2,2-dimethylpiperazine using General Procedure 22A, to afford the desired product.

Step II:

[0186] A mixture of the appropriate nitrile (1 eq.), sodium azide (3 eq) and triethylamine hydrochloride (3 eq.) in DM F (0.1 to 0.5M) is heated in the microwave for 2 to 3 hours at 130°C. The cooled mixture is evaporated to dryness and dried under vacuum. The residue is purified by reverse phase chromatography, using 0 to 30% water in ACN gradient to afford the desired tetrazole.

General Procedure 25: Preparation of Alkyl tetrazoles Step I:

[0187] A mixture of the appropriate acid (1.0 equiv), in THF (0.05 - 1 M) is treated with CDI (1-2 equiv) and allowed to stir at T from 15 min to lh. The mixture is then treated with NH₄OH (1-50 equiv.) and the mixture is allowed to stir at RT for lh to overnight. Water is added, and the aq. phase is extracted with EtOAc or DCM. The combined organic phase is washed with brine, dried over MgS0₄, filtered and the filtrate evaporated under reduced pressure affording the title compound. Optionally, the product is purified by silica gel chromatography.

Step II:

[0188] A mixture of the amide prepared above (1.0 equiv) in DCM (0.05 to 1M) is treated with TFAA (1-2 equiv) and the mixture is allowed to stir at temperatures ranging from RT to reflux for 15 min to overnight. The mixture is treated with H₂0 and allowed to stir for 10 min and the layers are separated and the aq. phase is extracted with DCM (2-4X) and the combined extracts are washed (H₂0, brine), dried (Na₂S0₄ or MgS0₄) and the organic phase is concentrated in vacuo. The residue is optionally purified by silica gel chromatography, preparative HPLC (H20-MeCN- formic acid), or used as is in the next step.

Step III:

[0189] Option A. A mixture of the nitrile prepared in the previous step (1.0 equiv), NaN₃ (1.0 - 15 equiv.) Et₃NH-HCI (1-15 equiv) in DMF (0.05 to 1 M) is heated at 150°C in an oil bath, or optionally in a microwave for 10 min to 12h. Water is added and the mixture extracted with EtOAc or DCM. Opionally, water is added and the mixture is lyophyllised directly to afford the desired compound.

[0190] Option B. A mixture of the nitrile prepared in the previous step (1.0 equiv), (Bu₃Sn)₂0 (0.1 to 2 equiv), TMSN₃ (1.0 - 15 equiv.) in dioxane (0.05 to 1 M) is heated at 50 to 200°C either thermally or in a microwave. The reaction is concentrated and purified by preparative HPLC to afford the desired product.

General Procedure 26: Preparation of Sulfonylureas [0191] Option A: A mixture of the amine (1.0 equiv) and Et₃N or iPr₂NEt (1-3 equiv) in MeCN (0.05 to 1 M) is treated with CDI (1-2 equiv) and the mixture is allowed to stir at T for 30 min to 6h. The mixture is then treated with the appropriate sulfonamide (3-5 equiv) and DBU (3-5 equiv) and the mixture is allowed to stir at 60°C for 1 to 24h. The reaction is cooled to RT, diluted with NM P, filtered and purified by mass directed preparative HPLC.

[0192] Option B: A mixture of the amine (1.0 equiv) and Et₃N or iPr₂NEt (1-5 equiv) in DCM (0.1 to 1 M) is treated with the appropriate sulfonylisocyanate (1-3 equiv) and stirred at RT for 1 to 18h. The mixture is concentrated in vacuo, dissolved in NM P, filtered and purified by UV or mass directed reverse phase HPLC.

General Procedure 27: Preparation of Ureas and Carbamates

[0193] Method A. Step I: A solution of the appropriate amine (1 equiv) in DCM (0.05 to 1M) containing pyridine (5-10 equiv) is cooled to 0°C and treated with a solution of phosgene (1-2 equiv, 15% w/w in toluene). The mixture is allowed to stir for times ranging from 30 min to overnight and the mixture is treated with Hunig's base (2-5 equiv) followed by the amine, amine salt, or alcohol (1-2 equiv) and the resulting mixture is allowed to stir at RT until judged complete by TLC or UPLCMS analysis (30 min to overnight). Standard aqueous workup affords the crude product which is of sufficient purity for use in subsequent steps (if necessary). In reactions affording final products, these are purified by mass directed reverse phase preparitve HPLC.

[0194] Step II (optional): In cases where the product contains an ester, it is hydrolysed using the methods in either General Procedures 11 or 23.

[0195] Method B. A mixture of the amine (1.0 equiv) and Et₃N or iPr₂NEt (1-3 equiv) in MeCN (0.05 to 1 M) is treated with CDI (1-2 equiv) and the mixture is allowed to stir at RT for 30 min to 6h. The mixture is then treated with the appropriate amine (3-5 equiv) and DBU (3-5 equiv) and the mixture is allowed to stir at 60°C for 1 to 24h. The reaction is cooled to RT, diluted with NMP, filtered and purified by mass directed preparative HPLC.

General Procedure 28: Preparation of Acylsulfonamides

[0196] A mixture of the carboxylic acid (1 equiv) in DCM (0.01 to 1 M) is sequentially treated with DMAP (1-4 equiv), EDCI (1-3 equiv) and the appropriate sulfonamide (1-3 equiv) at RT. The mixture is allowed to stir overnight (12-24 h) at RT and concentrated in vacuo. Purification by reverse phase HPLC affords the pure compounds. General Procedure 29: Preparation of squarate derivatives:

[0197] Option A: Preparation of squaric carbamates:

[0198] A solution of the appropriate piperazine analogue (1.0 equiv) in EtOH (0.1 to 1M) is treated with NEt₃ (1-3 equiv) followed by 3,4-diethoxycyclobut-3-ene-l,2-dione (2-4 equiv). The reaction mixture is heated at reflux until judged complete by TLC or LCMS. The solvents are removed in vacuo and the residue is triturated with Et₂0 to afford the crude product which is suitable for use in subsequent reactions. In cases where the product is a final product, the crude material is dissolved in NMP or DMSO and purified by mass or UV directed reverse HPLC.

[0199] Option B: Preparation of squaric urea analogues:

[0200] Step I: A solution of the appropriate squaric carbamate analogue (1.0 equiv) in EtOH (0.1 to 1M) is treated with NEt₃ (5-10 equiv) followed by the appropriate amine, or amine salt (4-10 equiv). The reaction mixture is heated at temperatures ranging from 50 to 80°C until judged complete by TLC or LCMS (12 to 36h). The reaction is diluted with NM P or DMSO and purified by mass or UV directed reverse HPLC. In some cases a side-product of hydrolysis results during the reaction. These analogues were isolated from the purification.

[0201] Step II (optional): In cases where the product contains an ester, it is hydrolysed using the methods in either General Procedures 11, 13 or 23.

General Procedure 30: Preparation of carboxylic acid glucuronide derivatives: [0202] A mixture of the appropriate acid (1 equiv), allyl (3S,4S,6R)-3,4,5,6- tetrahydroxytetrahydropyran-2-carboxylate (1.5 to 3 equiv), PPh₃ (1-3 equiv) in THF (0.05 to 0.5 M) was cooled in an ice bath and treated dropwise with DIAD (1 to 3 equiv). After stirring overnight, the mixure is treated with Pd(PPh₃)₄ (0.01 to 0.1 equiv) and pyrrolidine and stirring is continued at RT until the reaction is judged complete by LCMS (2 to 8h). The solvent is removed in vacuo and the residue is dissolved in solvents chosen from NMP, MeCN, DMSO, MeOH or DMF and purified by UV or mass directed reverse phase HPLC.

List of Intermediates

Preparation of Intermediates:

Preparation of Intermediate A: 7-tert-Butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2- carboxylic acid

Scheme 1

Step I: 4-tert-Butyl-6-(4-chloro-3-fluoro-phenyl)-2-hydroxy-pyridine-3-carbonitrile

[0203] Using l-(4-chloro-3-fluoro-phenyl)ethanone (11.1 g, 64.3 mmol) as the starting material and following General Procedure 4, the title compound (7.16 g, 47% yield) is obtained as a yellow solid after a final trituration in MeOH (40 mL). ^XH NM (400 M Hz, CDCI₃) δ 7.73 (dd, J = 8.6, 2.1 Hz, 1H), 7.69 - 7.61 (m, 2H), 6.70 (s, 1H), 1.55 (s, 9H). ¹⁹F NMR (376 MHz, CDCI₃) δ -112.44 (dd, J = 9.6, 7.3 Hz). ESI-MS m/z calc. 304.07788, found 305.24 (M+l)⁺; Retention time: 1.79 minutes using method C.

Step II: 4-tert-Butyl-6-(4-chloro-3-fluoro-phenyl)-2-iodo-pyridine-3-carbonitrile

[0204] Using 4-ieri-butyl-6-(4-chloro-3-fluoro-phenyl)-2-hydroxy-pyridine-3-carbonitrile (6.59 g, 21.6 mmol) as the starting material in MeCN (66 mL) and following General Procedure 5 using triflic acid (2.30 mL, 25.94 mmol) in the second step, the crude product is obtained. The residue is purified by flash chromatography on silica gel eluting with 0-60% DCM: Hexanes. The title compound (4.52 g, 50% yield) is obtained as a white solid. ^XH NMR (400 MHz, CDCI₃) δ 7.85 (dd, J = 10.1, 2.1 Hz, 1H), 7.74 (ddd, J = 8.4, 2.1, 0.8 Hz, 1H), 7.71 (s, 1H), 7.52 (dd, J = 8.4, 7.4 Hz, 1H), 1.56 (s, 9H). ¹⁹F NMR (376 M Hz, CDCI₃) δ -113.81 (dd, J = 10.1, 7.4 Hz). ESI-MS m/z calc. 413.97958, found 415.2 (M+l)⁺; Retention time: 3.03 minutes using method B.

Step III: 4-tert-Butyl-6-(4-chloro-3-fluoro-phenyl)-2-iodo-pyridine-3-carboxamide

[0205] Using 4-tert-butyl-6-(4-chloro-3-fluoro-phenyl)-2-iodo-pyridine-3-carbonitrile (4.47 g, 10.8 mmol) as the starting material and following General Procedure 7, the reaction mixture is neutralized using potassium carbonate in water (27.0 g in 170 mL). The resulting white precipitate is extracted with CHCI₃/iPrOH mixture (4:1, 3 x 120 mL). The combined organic extracts are washed with brine (60 mL), dried over MgS0₄, filtered and concentrated, affording the title compound (4.64 g, 99% yield) as an off-white solid. ^XH NMR (400 M Hz, CDCI₃) δ 7.79 (dd, J = 10.2, 2.0 Hz, 1H), 7.71 - 7.65 (m, 2H), 7.49 (dd, J = 8.4, 7.4 Hz, 1H), 6.01 (broad s, 1H), 5.78 (broad s, 1H), 1.49 (s, 9H). ¹⁹F NM R (376 MHz, CDCI₃) δ -114.49 (dd, J = 10.2, 7.4 Hz). ESI-MS m/z calc. 431.99017, found 433.22 (M+l)⁺; Retention time: 1.89 minutes using method C.

Step IV: 4-tert-Butyl-6-(4-chloro-3-fluoro-phenyl)-2-iodo-pyridin-3-amine

[0206] Using 4-tert-butyl-6-(4-chloro-3-fluoro-phenyl)-2-iodo-pyridine-3-carboxamide (1.00 g, 2.31 mmol) as the starting material and following General Procedure 8, the title compound (915 mg, 98% yield) is obtained as a light brown solid which is used in the next step without purification. XH NM (400 MHz, CDCI₃) δ 7.70 (dd, J = 10.6, 2.1 Hz, 1H), 7.60 (ddd, J = 8.4, 2.1, 0.8 Hz, 1H), 7.44 (s, 1H), 7.40 (dd, J = 8.3, 7.6 Hz, 1H), 4.51 (broad s, 2H), 1.45 (s, 9H). ¹⁹F NMR (376 M Hz, CDCI₃) δ -115.55 (dd, J = 10.6, 7.5 Hz). ESI-MS m/z calc. 403.99524, found 405.21 (M+l)⁺; Retention time: 2.52 minutes using method C.

Step V: 4-tert-Butyl-6-(4-chloro-3-fluoro-phenyl)-2-iodo-pyridin-3-ol

[0207] The reaction is performed using 4-ieri-butyl-6-(4-chloro-3-fluoro-phenyl)-2-iodo-pyridin-3- amine (2.27 g, 5.61 mmol) as the starting material and following General Procedure 9 with isopentyl nitrite (2.01 g, 17.1 mmol). After the hydrolysis step, the reaction mixture is diluted with EtOAc and H₂0 (10 mL each) and the layers are separated. The aqueous layer is extracted with EtOAc (2 x 50 mL). The combined organic extracts are washed with brine (50 mL), dried over MgS0₄, filtered and concentrated under reduced pressure. The residue is purified by flash chromatography on silica gel eluting with 0-60% DCM : Hexanes. The title compound (1.17 g, 51% yield) is obtained as a white solid. ^XH NM R (400 MHz, CDCI₃) δ 7.72 (dd, J = 10.5, 2.0 Hz, 1H), 7.65 - 7.58 (m, 1H), 7.47 (s, 1H), 7.46 - 7.39 (m, 1H), 5.65 (s, 1H), 1.43 (s, 9H). ¹⁹F NM R (376 M Hz, CDCI₃) δ -115.16 (dd, J = 10.4, 7.6 Hz). ESI-MS m/z calc. 404.97928, found 406.19 (M+l)⁺; Retention time: 1.75 minutes using method I.

Step VI: Ethyl 7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carboxylate

[0208] Using 4-ieri-butyl-6-(4-chloro-3-fluoro-phenyl)-2-iodo-pyridin-3-ol (1.21 g, 2.98 mmol) as the starting material and following General Procedure 10, the reaction mixture is poured carefully into a flask containing IN HCI (42 mL) and EtOAc (75 mL). The layers are separated and the aqueous layer is extracted with EtOAc (2 x 25 mL). The combined organic extracts are washed with half-saturated brine (50 mL), dried over MgS0₄ and filtered through Celite^®, rinsing with EtOAc and concentrated. The residue is purified by flash chromatography on silica gel eluting with 0-10% EtOAc: Hexanes. The mixed fractions are combined, concentrated and purified again by flash chromatography on silica gel eluting with 0-60% DCM: Hexanes. The title compound (408 mg, 36% yield) is obtained as a pale yellow solid. ^XH NMR (400 M Hz, CDCI₃) δ 7.85 (dd, J = 10.4, 2.0 Hz, 1H), 7.74 (dd, J = 8.4, 1.7 Hz, 1H), 7.67 (s, 1H), 7.61 (s, 1H), 7.56 - 7.46 (m, 1H), 4.47 (q, J = 7.1 Hz, 2H), 1.58 (s, 9H), 1.45 (t, J = 7.1 Hz, 3H). ¹⁹F NM R (376 M Hz, CDCI₃) δ -115.06 (dd, J = 10.3, 7.6 Hz). ESI-MS m/z calc. 375.10376, found 376.02 (M+l)⁺; Retention time: 1.64 minutes using method I.

Step VII: 7-tert-Butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carboxylic acid [0209] Using ethyl 7-ieri-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carboxylate (398 mg, 1.06 mmol) as the starting material and following General Procedure 11, the reaction mixture is neutralized with IN HCI (2.2 mL), and diluted with DCM (20 mL) and H₂0 (10 mL). The layers are separated. The aqueous layer is further diluted with H₂0 (10 mL), acidified with IN HCI (0.2 mL) and extracted with DCM (20 mL, 2 x 10 mL). The combined organic extracts are dried over MgS0₄, filtered and concentrated, affording the title compound (417 mg, 100% yield) as a pale yellow solid. ^XH NM (400 M Hz, CDCI₃ and DMSO-d6) δ 7.72 (dd, J = 10.4, 2.0 Hz, 1H), 7.61 (dd, J = 8.4, 2.1 Hz, 1H), 7.51 (s, 1H), 7.47 (s, 1H), 7.41 - 7.32 (m, 1H), 1.44 (s, 9H). ¹⁹F NM R (376 MHz, CDCI₃ and DMSO-d6) δ -115.37 (dd, J = 10.4, 7.6 Hz). ESI-MS m/z calc. 347.07245, found 348.34 (M+l)⁺; Retention time: 1.18 minutes using method I.

Preparation of Intermediate B: 7-(tert-butyl)-5-(4-chlorophenyl)furo[3,2-blpyridine-2-carboxylic acid

Scheme 2

Step I: 4-tert-Butyl-6-(4-chlorophenyl)-2-hydroxy-pyridine-3-carbonitrile

[0210] Using l-(4-chlorophenyl)ethanone (18.50 g, 119.7 mmol) as the starting material and following General Procedure 4, the title compound is obtained as a pink solid (9.09 g, 34% yield) after a final trituration in hot EtOH. ^XH NM R (400 MHz, Chloroform-d) δ 7.88 - 7.78 (m, 2H), 7.61 - 7.54 (m, 2H), 6.70 (s, 1H), 1.55 (s, 9H). ESI-MS m/z calc. 286.08728, found 287.35 (M+l)⁺; Retention time: 1.72 minutes using method C.

Step II: 4-tert-Butyl-6-(4-chlorophenyl)-2-iodo-pyridine-3-carbonitrile

[0211] Using 4-ieri-butyl-6-(4-chlorophenyl)-2-hydroxy-pyridine-3-carbonitrile (9.07 g, 31.6 mmol) as the starting material and following General Procedure 5 using triflic acid (3.36 mL, 38.0 mmol) in the second step, the crude product is obtained. The residue is purified by flash chromatography on silica gel eluting with 0-80% DCM: Hexanes. The title compound (6.58 g, 52% yield) is obtained as a light yellow solid. ^XH NMR (400 MHz, CDCI₃) δ 8.00 - 7.90 (m, 2H), 7.72 (s, 1H), 7.52 - 7.41 (m, 2H), 1.56 (s, 9H). ESI-MS m/z calc. 395.98902, found 397.22 (M+l)⁺; Retention time: 2.93 minutes using method B.

Step III: 4-tert-Butyl-6-(4-chlorophenyl)-2-iodo-pyridine-3-carboxamide

[0212] Using 4-ieri-butyl-6-(4-chlorophenyl)-2-iodo-pyridine-3-carbonitrile (6.60 g, 16.6 mmol) as the starting material and following General Procedure 7, the reaction mixture is neutralized using potassium carbonate in water (42.00 g in 250 mL). The resulting white precipitate is extracted with CHCI₃//PrOH mixture (4:1, 3 x 100 mL). The combined organic extracts are washed with H₂0 (100 mL), brine (100 mL), dried over MgS0₄, filtered and concentrated, affording the title compound (6.97 g, 100% yield) as an off-white solid. ¹H NMR (400 MHz, CDCI₃) δ 7.94 - 7.83 (m, 2H), 7.70 (s, 1H), 7.49 - 7.39 (m, 2H), 6.03 (broad s, 1H), 5.80 (broad s, 1H), 1.49 (s, 9H). ESI-MS m/z calc. 413.99957, found 415.25 (M+l)⁺; Retention time: 1.9 minutes using method C.

Step IV: 4-tert-Butyl-6-(4-chlorophenyl)-2-iodo-pyridin-3-amine

[0213] Using 4-ieri-butyl-6-(4-chlorophenyl)-2-iodo-pyridine-3-carboxamide (3.03 g, 7.31 mmol) as the starting material and following General Procedure 8, the title compound (2.66 g, 94% yield) is obtained as a dark purple solid which is used in the next step without purification. ^XH NMR (400 MHz, CDCI₃) δ 7.84 - 7.77 (m, 2H), 7.45 (s, 1H), 7.39 - 7.34 (m, 2H), 4.45 (s, 2H), 1.45 (s, 9H). ESI-MS m/z calc. 386.00467, found 387.23 (M+l)⁺; Retention time: 2.46 minutes using method C.

Step V: 4-tert-Butyl-6-(4-chlorophenyl)-2-iodo-pyridin-3-ol

[0214] Using 4-ieri-butyl-6-(4-chlorophenyl)-2-iodo-pyridin-3-amine (2.99 g, 7.73 mmol) as the starting material and following General Procedure 9, the reaction mixture after the hydrolysis step is diluted with EtOAc and H₂0 (150 mL each) and the layers are separated. The aqueous layer is extracted with EtOAc (2 x 75 mL). The combined organic extracts are washed with brine (150 mL), dried over MgS0₄, filtered and concentrated. The residue is purified by flash chromatography on silica gel eluting with 0-30% DCM : Hexanes. The title compound (2.35 g, 78% yield) is obtained as a white solid. ^XH NM (400 M Hz, CDCI₃) δ 7.85 - 7.79 (m, 2H), 7.48 (s, 1H), 7.43 - 7.36 (m, 2H), 5.60 (broad s, 1H), 1.43 (s, 9H). ESI-MS m/z calc. 386.98868, found 388.16 (M+l)⁺; Retention time: 1.74 minutes using method I.

Step VI: Ethyl 7-tert-butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2-carboxylate

[0215] Using 4-ferf-butyl-6-(4-chlorophenyl)-2-iodo-pyridin-3-ol (3.13 g, 8.07 mmol) as the starting material and following General Procedure 10, the reaction mixture is poured carefully into a flask containing IN HCI (120 mL) and EtOAc (100 mL). The layers are separated and the aqueous layer is extracted with EtOAc (100 mL, 2 x 50 mL). The combined organic extracts are washed with half-saturated brine (100 mL), dried over MgS0₄ and filtered through Celite^®, rinsing with EtOAc and concentrated. The residue is purified by flash chromatography on silica gel eluting with 0-10% EtOAc: Hexanes. The title compound (1.08 g, 37% yield) is obtained as a pale yellow solid. ^XH NM R (400 MHz, CDCI₃) δ 8.00 - 7.86 (m, 2H), 7.68 (s, 1H), 7.62 (s, 1H), 7.53 - 7.39 (m, 2H), 4.47 (q, J = 7.1 Hz, 2H), 1.58 (s, 9H), 1.45 (t, J = 7.1 Hz, 3H). ESI-MS m/z calc. 357.11316, found 358.61 (M+l)⁺; Retention time: 1.84 minutes using method I.

Step VII: 7-tert-Butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2-carboxylic acid (Intermediate B)

[0216] Using ethyl 7-ieri-butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2-carboxylate (1.06 g, 2.96 mmol) as the starting material and following General Procedure 11, the reaction mixture is neutralized with IN HCI (6 mL) and extracted with DCM (20 mL, 2 x 10 mL). The combined organic extracts washed with brine (10 mL), and the brine layer is further extracted with DCM (2 x 10 mL). The combined organic extracts are dried over MgS0₄, filtered and concentrated, affording the title compound (1.14 g, 100% yield) as a pale yellow solid. ^XH NM R (400 MHz, CDCI₃ and DMSO-d6) δ 7.81 - 7.67 (m, 2H), 7.44 (s, 1H), 7.42 (s, 1H), 7.32 - 7.20 (m, 2H), 1.38 (s, 9H). ESI-MS m/z calc. 329.08188, found 330.31 (M+l)⁺; Retention time: 1.07 minutes using method I.

Preparation of Intermediate C: 7-(tert-butyl)-5-(4,4-dimethylcvclohexyl)furo[3,2-blpyridine-2- carboxylic acid

Scheme 3

Step I: (5-chlorofuro[3,2-b]pyridin-2-yl)methanol

[0217] To a round bottom flask containing a solution of 6-chloro-2-iodo-pyridin-3-ol (50.0 g, 196 mmol) in 1,4-dioxane (150.0 mL) is added DIPEA (63.2 g, 85.2 mL, 489.2 mmol), PdCI₂(PPh₃)₂ (6.87 g, 9.79 mmol) and Cul (3.73 g, 19.6 mmol). A solution of prop-2-yn-l-ol (14.3 g, 14.8 mL, 254 mmol) in 1,4-dioxane (100 mL) is added dropwise over 45 minutes at 75°C and stirring is pursued for 18h. The solution is cooled to room temperature, filtered on a Celite pad and the filtrate concentrated under reduced pressure. The filtrate is diluted with water and ethyl acetate and the phases are separated. The aqueous phase is washed twice with ethyl acetate and the combined organic phase is washed with brine, dried over MgS0₄, filtered and the filtrate evaporated under reduced pressure. The residue is purified by silica gel chromatography affording the title compound as a tan solid. (5-chlorofuro[3,2-b]pyridin-2-yl)methanol (27.04 g, 147.3 mmol, 75.3%) ^XH NM (400 MHz, CDCI₃) δ 7.66 (d, J = 8.6 Hz, 1H), 7.20 (d, J = 8.6 Hz, 1H), 6.81 (s, 1H), 4.81 (d, J = 6.2 Hz, 2H), 2.26 (t, J = 6.3 Hz, 1H). ESI-MS m/z calc. 183.00871, found 184.16 (M+l)⁺; Retention time: 0.86 minutes using method C.

Step II: (7-(tert-butyl)-5-(4,4-dimethylcyclohex-l-en-l-yl)furo[3,2-b]pyridin-2-yl)methanol

[0218] A mixture of (7-tert-butyl-5-chloro-furo[3,2-b]pyridin-2-yl)methanol (100 g, 417 mmol), 2- (4,4-dimethylcyclohexen-l-yl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (108.4 g, 458.9 mmol), and Na₂C0₃ (134.6 g, 1.270 mol) is suspended in dioxane (2 L) and water (667 mL). The resulting solution is degassed with nitrogen for 5 minutes prior to adding [1,1'- Bis(diphenylphosphino)ferrocene]dichloropalladium(ll) complex with dichloromethane (17.04 g, 20.86 mmol). It is then stirred at 90°C for 18h. The solvent is distilled, and to the cooled residue is water (500 mL) and EtOAc (2.5 L). The biphasic mixture is filtered through Celite and the layers are separated. The aqueous layer is back-extracted EtOAc (1000 mL). The combined organic layers are washed with brine then dried over MgS0₄, filtered then concentrated under reduced pressure to approximately 2L. The residue is treated with a solution of HCI in dioxane (4.0 M, 160 mL, 640 mmol). Heptane (300.0 mL) is added and the mixture is stirred at T for 16h. The solid is filtered, washed with EtOAc and dried under reduced pressure to afford the title compound (118.5g, 81%) as a brown-beige solid, which is used directly in the next step. ESI-MS m/z calc. 313.2042, found 313.76 (M+l)⁺; Retention time: 1.55 minutes using method C.

Step III: 7-(tert-butyl)-5-(4,4-dimethylcyclohex-l-en-l-yl)furo[3,2-b]pyridine-2-carboxylic acid

[0219] To a mixture of NMO hydrate (118.3 g, 875.2 mmol) and TPAP (2.28 g, 6.49 mmol) in acetonitrile (710 mL) is added portionwise (7-(tert-butyl)-5-(4,4-dimethylcyclohex-l-en-l- yl)furo[3,2-b]pyridin-2-yl)methanol (38.7 g, 101 mmol) while maintaining the temperature below 35^QC. The reaction mixture is stirred at room temperature for lh. The solution is cooled to 0°C, /^'- PrOH (78 mL, 1.0 mol) is added slowly and the reaction is stirred for 15 minutes. The reaction mixture is diluted with water and the volatiles are removed under reduced pressure. The aq. phase is basified to pH 13 using 2N NaOH and extracted with MTBE three times. The combined organic phases are concentrated under reduced pressure and the residue is diluted with water (535 mL). The resulting aqueous solution is acidified to pH 3 with 2N HCI, the product formed is collected by filtration, washed with water and dried under reduced pressure affording the title compound (20.03 g, 60%) as a beige solid, which is used directly in the next step. ESI-MS m/z calc. 327.1834, found 328.2 (M+l)⁺; Retention time: 3.47 minutes using method A.

Step IV: 7-(tert-butyl)-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridine-2-carboxylic acid (Intermediate C)

[0220] A Buchi pressure vessel purged with N₂ is charged with Pd on carbon (wet base, 5.11 g, 4.80 mmol) followed by a solution of 7-tert-butyl-5-(4,4-dimethylcyclohexen-l-yl)furo[3,2-b]pyridine- 2-carboxylic acid (15.59 g, 45.66 mmol) and AcOH (5.5 mL, 97 mmol) in THF (100 mL) and then ethanol (400 mL). Nitrogen atmosphere is evacuated under reduced pressure and re-filled with hydrogen 3 times and the suspension is stirred under 80 psi hydrogen pressure at room temperature for 4 days. The suspension is filtered over celite, washed with THF and the volatiles are removed under reduced pressure. This solid is suspended MeOH: Water (1:1) then heated to reflux. The mixture is cooled to RT then stirred for lhr in an ice-bath (5-10^QC). The product formed is collected by filtration, washed with water then heptane, and dried under reduced pressure affording the title compound (12.60 g, 80%) as a brown-beige solid, which is used directly in the next step. ESI-MS m/z calc. 329.1991, found 329.77 (M+l)⁺; Retention time: 3.26 minutes using method A.

Preparation of Intermediate D: [7-tert-Butyl-5-(4-chloro-3-fluorophenyl)furo[3,2-blpyridin-2-yll-(2,2- dimethylpiperazin-l-yl)methanone hydrochloride

Step I: tert-Butyl 4-[7-tert-butyl-5-(4-chloro-3-fluorophenyl)furo[3,2-b]pyridine-2-carbonyl]- 3,3-dimethyl-piperazine-l-carboxylate

[0221] The intermediate is prepared according to General Procedure 1 using a solution of Intermediate A (10.0 g, 28.3 mmol) in DMF (110 mL), HATU (12.0 g, 31.6 mmol), ferf-butyl 3,3- dimethylpiperazine-l-carboxylate (6.64, 31.0 mmol) and DIPEA (12.5 ml, 71.8 mmol) affording the title compound (15.38 g, quantitative yield) as a pale yellow solid, which is used directly in the next step. 1H NMR (400 M Hz, DMSO-d₆) δ 8.16 (dd, J = 11.0, 2.0 Hz, 1H), 8.07 - 7.97 (m, 1H), 7.82 (s, 1H), 7.76 - 7.64 (m, 1H), 7.51 (s, 1H), 3.85 (s, 2H), 3.57 - 3.38 (m, 4H), 1.52 (s, 10H), 1.48 (s, 6H), 1.42 (s, 9H). ESI-MS m/z calc. 543.2300, found 546.10 (M+l)⁺; Retention time: 1.23 minutes using method J.

Step II: [7-tert-Butyl-5-(4-chloro-3-fluorophenyl)furo[3,2-b]pyridin-2-yl]-(2,2- dimethylpiperazin-l-yl)methanone hydrochloride (Intermediate D)

[0222] Using ferf-butyl 4-[7-ferf-butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-piperazine-l-carboxylate (15.38 g, 28.27 mmol) as the starting material and following General Procedure 2 with the exception that the reaction is performed in dioxane instead of DCM and using HCI in dioxane (35 mL of 4 M, 140 mmol). The title compound (12.80 g, 88% yield) is obtained as a pale yellow solid, which is used directly in the next step. 1H NM R (400 M Hz, DMSO-d₆) δ 9.37 (broad s, 2H), 8.17 (dd, J = 11.0, 1.9 Hz, 1H), 8.03 (dd, J = 8.4, 1.8 Hz, 1H), 7.85 (s, 1H), 7.72 (t, J = 8.2 Hz, 1H), 7.62 (s, 1H), 3.90 - 3.74 (m, 2H), 3.38 - 3.27 (m, 2H), 3.26 - 3.15 (m, 2H), 1.58 (s, 6H), 1.53 (s, 9H). ESI-MS m/z calc. 443.1776, found 443.73 (M+l)⁺; Retention time: 0.66 minutes using method J.

Preparation of Intermediate E: (7-(tert-butyl)-5-(4,4-dimethylcvclohexyl)furo[3,2-blpyridin-2-yl)(2,2- dimethylpiperazin-l-vDmethanone hydrochloride

Scheme 4

Step I: tert-butyl 4-(7-(tert-butyl)-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridine-2-carbonyl)- 3,3-dimethylpiperazine-l-carboxylate

[0223] ferf-butyl 3,3-dimethylpiperazine-l-carboxylate (19.90 g, 92.86 mmol) and (2,2- dimethylpiperazin-l-yl)methanone hydrochloride (Intermediate C) (27.87 g, 84.60 mmol) are dissolved in DM F (300 mL) before HATU (35.45 g, 93.23 mmol) and DIPEA (27.72 g, 37.36 mL, 214.5 mmol) are added. The solution is stirred at room temperature for 3h, and then water is added dropwise. The aq. phase is extracted three times with EtOAc. The combined organic phase is washed with water and brine. The organic phase is then dried over Na₂S0₄, filtered and the filtrate evaporated under reduced pressure. The crude product is purified passing through a pad of silica gel (25%-33% EtOAc/hexanes affording the title product (34.28 g, 91% yield) as a yellow orange foamy solid, which is used directly in the next step. 1H NM R (400 MHz, Chloroform-d) δ 7.34 (s, 1H), 7.06 (s, 1H), 3.84 (dd, J = 6.6, 4.8 Hz, 2H), 3.67 - 3.45 (m, 4H), 2.71 (tt, J = 10.3, 5.5 Hz, 1H), 1.85 - 1.77 (m, 4H), 1.77 - 1.71 (m, 1H), 1.57 - 1.51 (m, 2H), 1.49 (s, 9H), 1.49 (s, 9H), 1.43 - 1.31 (m, 3H), 1.01 (s, 3H), 0.97 (s, 3H). ESI-MS m/z calc. 525.3567, found 526.61 (M+l)⁺; Retention time: 3.15 minutes using method A.

Step II: (7-(tert-butyl)-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridin-2-yl)(2,2- dimethylpiperazin-l-yl)methanone hydrochloride (Intermediate E)

[0224] Using tert-butyl 4-[7-tert-butyl-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridine-2-carbonyl]- 3,3-dimethyl-piperazine-l-carboxylate (34.2 g, 65.1 mmol) as the starting material and following General Procedure 2 with the exception that the reaction is performed in dioxane instead of DCM and using HCI in dioxane (162.6 mL of 4.0 M, 650.5 mmol). The title compound (31.4 g, quantitative) is obtained as an orange foam, which is used directly in the next step. ESI-MS m/z calc. 425.30423, found 426.53 (M+l)⁺; Retention time: 3.14 minutes using method A.

Preparation of Intermediate F: [7-tert-Butyl-5-(4-chlorophenyl)furo[3,2-blpyridin-2-yll-(2,2- dimethylpiperazin-l-vPmethanone hydrochloride

Scheme 5

Step I: tert-Butyl 4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-piperazine-l-carboxylate

[0225] The intermediate is prepared according to General Procedure 1 using a solution of Intermediate B (396 mg, 1.20 mmol) in DMF (5 mL), HATU (548 mg, 1.441 mmol), ferf-butyl 3,3- dimethylpiperazine-l-carboxylate (283 μί, 1.32 mmol) and DIPEA (732 μί, 4.20 mmol) affording the title compound (609 mg, 96% yield) as a beige solid, which is used directly in the next step. ESI-MS m/z calc. 525.23944, found 526.55 (M+l)⁺; Retention time: 3.26 minutes using method B.

Step II: [7-tefNButyl-5-(4-chlorophenyl)furo[3,2-b]pyridin-2-yl]-(2,2-dimethylpiperazin-l- yl)methanone hydrochloride (Intermediate F)

[0226] Using ferf-butyl 4-[7-ferf-butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-piperazine-l-carboxylate (609 mg, 1.16 mmol) as the starting material and following General Procedure 2 with the exception that the reaction is performed without DCM and using HCI in dioxane (8.7 mL of 4 M, 35 mmol). The title compound (642 mg, 100% yield) is obtained as a pale yellow solid, which is used directly in the next step. ESI-MS m/z calc. 425.187, found 426.45 (M+l)⁺; Retention time: 1.77 minutes using method C.

Preparation of Intermediate G: ethyl 2-(3,3-dimethylpiperazin-l-yl)-5-methyl-pyridine-4-carboxylate

[0227] A solution of 2,2-dimethylpiperazine (132 mg, 1.16 mmol), ethyl 2-chloro-5-methyl-pyridine- 4-carboxylate (210 mg, 1.05 mmol) and N-ethyl-N-isopropyl-propan-2-amine (193 μί, 1.11 mmol) in DMSO (2.5 mL) is heated at 120°C in microwave for 8h, then for one day at 150°C thermally. To the reaction mixture is added water, extracted three times with EtOAc, dried over sodium sulfate, filtered, and the solvent removed under reduced pressure. The residue is used in the next reaction without further purification or purification. ESI-MS m/z calc. 277.17902, Found: 278.73 Retention time: 1.20 minutes using Method L.

Preparation of Intermediate H. methyl 2-(3,3-dimethylpiperazin-l-yl)-6-methyl-pyridine-4- carboxylate (Dihydrochloride salt)

[0228] Step I: To a solution of tert-butyl 2,2-dimethylpiperazine-l-carboxylate (200 mg, 0.933 mmol) in DM F (3 mL) are added methyl 2-bromo-6-methyl-pyridine-4-carboxylate (236.3 mg, 1.03 mmol), K₂C0₃ (258 mg, 1.87 mmol), X-PHOS (89.0 mg, 0.187 mmol) and Pd₂(dba)₃ (85.46 mg, 0.0933 mmol) in DMF (3 mL). After degassing for 5 min, The mixture is stirred at 90°C under nitrogen overnight. The mixture is diluted with EtOAc (60 mL), washed with water and brine consecutively, dried over sodium sulfate, filtered. The filtrated is concentrated to dryness under reduced pressure. The residue is purified on Biotage SNAP 25g silica gel cartridge eluting with EtOAc/hexanes 0-30% in 20 CV to obtain a yellowish solid, (94 mg, 28%), which will be used directly in the next step without further purification. LCMS: 364.30 (M+H)+; calc. 364.2236. RT: 1.78 min using method C.

[0229] Step II: To the mixture from step 1 tert-butyl 4-(4-methoxycarbonyl-6-methyl-2-pyridyl)-2,2- dimethyl-piperazine-l-carboxylate (94 mg) in DCM (1 mL) is added 4M HCI/dioxane (500 μΐ of 4 M, 2.00 mmol). The mixture is stirred at rt for 2h. Then the volatiles are removed under reduced pressure to afford methyl 2-(3,3-dimethylpiperazin-l-yl)-6-methyl-pyridine-4-carboxylate (Dihydrochloride salt) (86 mg, 99%) as an off-white solid. LCMS: 264.73 (M+H)+; calc. 264.1712. T: 0.68 min using Method C.

Preparation of Intermediate I. 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2- carbonyll-3,3-dimethyl-piperazin-2-one

[0230] 7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carboxylic acid (3.00 g, 8.40 mmol) is dissolved in DM F (30 mL) and DIPEA (5.80 mL, 33.3 mmol) followed by 3,3- dimethylpiperazin-2-one (1.18 g, 9.21 mmol) are added. It is stirred 2 min before addition of HATU (4.79 g, 12.6 mmol) in a single portion. The reaction mixture is stirred at RT for lh. Water is slowly added and the mixture stirred for 2 hours before being filtered. The solid is allowed to dry under suction ON (18h), giving 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2- b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-2-one (3.80 g, 93%). ^XH NMR (400 MHz, DMSO- d₆) δ 8.20 - 8.17 (m, 1H), 8.15 (dd, J = 11.0, 2.1 Hz, 1H), 8.04 - 7.98 (m, 1H), 7.81 (s, 1H), 7.74 - 7.64 (m, 1H), 7.57 (s, 1H), 3.76 - 3.69 (m, 2H), 3.40 - 3.32 (m, 2H), 1.67 (s, 6H), 1.49 (s, 9H). ESI- MS m/z calc. 457.15686, found 458.2 (M+l)+; ESI-MS m/z calc. 457.15686, found 458.2 (M+l)+; Retention time: 0.98 minutes using Method J.

Preparation of Intermediate J. 4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2-blpyridine-2-carbonyll-3,3- dimethyl-piperazin-2-one

[0231] 7-tert-butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2-carboxylic acid (3.00 g, 9.10 mmol) is dissolved in DM F (30 mL) and DIPEA (6.30 mL, 36.2 mmol) followed by 3,3-dimethylpiperazin-2- one (1.28 g, 9.99 mmol) is added. After stirring for 2 min HATU (5.19 g, 13.65 mmol) is added in a single portion. The reaction mixture is stirred at RT for lh. Water is slowly added and the mixture is stirred for 2 hours before being filtered. The collected solid is allowed to dry overnight under suction. (3.84 g, 89%) ^XH NM (400 MHz, DMSO-d6) δ 8.20 - 8.16 (m, 1H), 8.15 - 8.10 (m, 2H), 7.75 (s, 1H), 7.56 (s, 1H), 7.56 - 7.50 (m, 2H), 3.77 - 3.69 (m, 2H), 3.40 - 3.32 (m, 2H), 1.67 (s, 6H), 1.49 (s, 9H). ESI-MS m/z calc. 439.16626, found 440.25 (M+l)+; Retention time: 0.95 minutes using Method J.

Preparation of Intermediate K. 4-[7-tert-butyl-5-(4,4-dimethylcvclohexyl)furo[3,2-blpyridine-2- carbonyll-3,3-dimethyl-piperazin-2-one

[0232] 7-tert-butyl-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridine-2-carboxylic acid (3.00 g, 8.75 mmol) is dissolved in DMF (30 mL) and DIPEA (6.10 mL, 35.0 mmol) followed by 3,3- dimethylpiperazin-2-one (1.23 g, 9.60 mmol) are added. HATU (4.99 g, 13.1 mmol) is added in a single protion. The reaction mixture is stirred at RT for lh and water is slowly added. The mixture is allowed to stir for 2 hours then filtered and allowed to dry under suction overnight. (3.8 g, 91%) ^XH NMR (400 MHz, DMSO-d6) δ 8.21 - 8.11 (m, 1H), 7.44 (s, 1H), 7.13 (s, 1H), 3.74 - 3.66 (m, 2H), 3.37 - 3.31 (m, 2H), 2.73 - 2.61 (m, 1H), 1.82 - 1.58 (m, 10H), 1.48 - 1.39 (m, 11H), 1.36 - 1.24 (m, 2H), 0.93 (d, J = 8.7 Hz, 6H). ESI-MS m/z calc: 439.28348, Found: 440.35; Retention time: 0.99 min using Method J.

Preparation of Intermediate L. [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridin-2-yll-

(2,2-dimethyl-l,4-diazepan-l-yl)methanone

Step I: tert-butyl 3,3-dimethyl-l,4-diazepane-l-carboxylate (Intermediate V)

[0233] In a 25 mL flask, TEA (728 μί, 5.22 mmol) is added to a stirred mixture of 2,2-dimethyl-l,4- diazepane (Dihydrochloride salt) (500 mg, 2.49 mmol) in DCM (5 mL). The solution is cooled to 0°C and a solution of di-tert-butyl dicarbonate (543 mg, 2.49 mmol) in DCM is then added dropwise. The reaction is stirred for 72h and is treated with water (stir 5 min) and the layers separated using a phase separator cartdrige. Aqueous phase is extracted again with DCM and the organic filtrate is concentrated under reduced pressure to give the desired product (580 mg, 102%) as a light yellow solid which is used as is in the next step. ESI-MS m/z calc. 228.18378, found 229.14 (M+l)+; Retention time: 0.56 minutes using Method M.

Step II: tert-butyl 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]- 3,3-dimethyl-l,4-diazepane-l-carboxylate

[0234] N-ethyl-N-isopropyl-propan-2-amine (1.33 mL, 7.61 mmol) is added to a stirred mixture of 7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carboxylic acid (661.1 mg, 1.90 mmol), tert-butyl 3,3-dimethyl-l,4-diazepane-l-carboxylate (435 mg, 1.91 mmol) and HATU (1.087 g, 2.86 mmol) in DM F (4.3 mL). The reaction is stirred 4 hours at rt, water is added along with EtOAc and phases are separated. The aqueous phase is extracted with EtOAc. The combined organic phases are washed (H₂0, brine), dried over sodium sulfate, filtered and evaporated under reduced pressure. The residue is purified on Isco Combiflash 40 g silica gel cartridge eluting with 0-100% EtOAc/Hexanes (loading with DCM). The desired fractions are combined and concentrated to give upon trituration in diethyl ether tert-butyl 4-[7-tert-butyl-5- (4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-l,4-diazepane-l- carboxylate (700 mg, 65%). ^ΧΗ NM R (400 M Hz, DMSO-d6) δ 8.21 - 8.13 (m, 1H), 8.07 - 7.99 (m, 1H), 7.81 (s, 1H), 7.76 - 7.67 (m, 1H), 7.45 (s, 1H), 3.69 (s, 1H), 3.56 (s, 2H), 3.51 - 3.43 (m, 2H), 3.29 (s, 1H), 1.93 (s, 2H), 1.59 - 1.48 (m, 15H), 1.43 (s, 9H). ESI-MS m/z calc. 557.24567, found 559.69 (M+l)+; Retention time: 5.15 minutes using Method A.

Step III: [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]-(2,2-dimethyl-l,4- diazepan-l-yl)methanone

[0235] To a stirred soltuion of tert-butyl 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2- b]pyridine-2-carbonyl]-3,3-dimethyl-l,4-diazepane-l-carboxylate (182 mg, 0.326 mmol) in DCM (1 mL), TFA (251.2 μί, 3.26 mmol) is added and the reaction is stirred at rt for 4 hours. TFA (126 μί, 1.63 mmol) is added and the reaction is stirred for 2 hours. The reaction mixture is concentrated under reduced pressure. Sat. NaHC0₃ (aq) is added along with DCM and the mixture is stirred for 5 minutes. The phases are separated using a phase separator and the filtrate is concentrated to give [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]- (2,2-dimethyl-l,4-diazepan-l-yl)methanone (143 mg, 96%) ^XH NM (400 MHz, DMSO-d6) δ 8.21 - 8.11 (m, 1H), 7.44 (s, 1H), 7.13 (s, 1H), 3.74 - 3.66 (m, 2H), 3.37 - 3.31 (m, 2H), 2.73 - 2.61 (m, 1H), 1.82 - 1.58 (m, 10H), 1.48 - 1.39 (m, 11H), 1.36 - 1.24 (m, 2H), 0.93 (d, J = 8.7 Hz, 6H). ESI- MS m/z calc. 457.19324, found 458.39 (M+l)+; Retention time: 0.68 minutes using Method J.

Preparation of Intermediate M. Methyl 2-cyano-6-(3,3-dimethylpiperazin-l-yl)pyridine-3-carboxylate

(Dihydrochloride salt).

[0236] Step I: To a solution of methyl 2,6-dichloropyridine-3-carboxylate (1.02 g, 4.95 mmol) in NM P (2 mL) are added tert-butyl 2,2-dimethylpiperazine-l-carboxylate (1.11 g, 5.18 mmol) and TEA (415 μί, 2.98 mmol). The mixture is stirred at rt for 1 day. Then it is diluted with water. The mixture is stirred at rt for 20 min, filtered. The solid is washed with water, dried in vacuo. The residue is purified on Biotage SNAP 50g silica gel cartridge eluting with EtOAc/hexanes 0-20% in 15 CV to obtain regioisomer 1 tert-butyl 4-(6-chloro-5-methoxycarbonyl-2-pyridyl)-2,2-dimethyl- piperazine-l-carboxylate (949 mg, 50%) as a white solid. ¹H NM R (400 M Hz, DMSO-d6) δ 8.02 (d, J = 8.9 Hz, 1H), 6.68 (d, J = 8.9 Hz, 1H), 3.84 - 3.74 (m, 7H), 3.51 (t, J = 5.7 Hz, 2H), 1.42 (s, 9H), 1.33 (s, 6H) and 2 tert-butyl 4-(6-chloro-3-methoxycarbonyl-2-pyridyl)-2,2-dimethyl-piperazine- 1-carboxylate (690 mg) as a white solid. ^XH NM R (400 MHz, DMSO-d6) δ 7.91 (d, J = 7.9 Hz, 1H), 6.78 (d, J = 7.9 Hz, 1H), 3.79 (s, 3H), 3.69 - 3.59 (m, 2H), 3.46 (s, 2H), 3.34 - 3.21 (m, 2H), 1.39 (s, 9H), 1.32 (s, 6H). A NOESY experiment confirms the regiochemistry of the compounds.

[0237] Step II: To a solution of tert-butyl 4-(6-chloro-5-methoxycarbonyl-2-pyridyl)-2,2-dimethyl- piperazine-l-carboxylate (500 mg, 1.30 mmol) in DM F (8 mL) are added dicyanozinc (153 mg, 1.30 mmol) and Pd(PPh₃)₄ (300 mg, 0.256 mmol). The mixture is degassed for 5 min and then stirred at 100°C under nitrogen for 2h. Then the volatiles are removed under reduced pressure. The residue is purified on Biotage SNAP 25g silica gel cartridge eluting with EtOAc/hexane 0-30% in 20 CV to obtain tert-butyl 4-(6-cyano-5-methoxycarbonyl-2-pyridyl)-2,2-dimethyl-piperazine- 1-carboxylate (430 mg, 88%) as a white solid. LC-MS(ESI): 375.33 (M+H)+; calc. 375.2032. RT: 1.8 min using Method C.

[0238] Step III: To a solution of tert-butyl 4-(6-cyano-5-methoxycarbonyl-2-pyridyl)-2,2-dimethyl- piperazine-l-carboxylate (430 mg) in DCM (5 mL) is added 4M HC/dioxane (3.0 mL of 4 M, 12 mmol). The mixture is stirred at rt for 2h. Then the volatiles are removed under reduced pressure to provide methyl 2-cyano-6-(3,3-dimethylpiperazin-l-yl)pyridine-3-carboxylate (Dihydrochloride salt) (370 mg, 93%) as a white solid. LC-MS(ESI): 275.33 (M+H)+; calc. 275.1508. RT: 0.64 min using Method C.

Preparation of Intermediate N. 6-cyano-2-(3,3-dimethylpiperazin-l-yl)pyridine-3-carboxylate

(Dihydrochloride salt)

Step I: tert-butyl 4-(6-cyano-3-methoxycarbonyl-2-pyridyl)-2,2-dimethyl-piperazine-l- carboxylate

[0239] To a solution of tert-butyl 4-(6-chloro-3-methoxycarbonyl-2-pyridyl)-2,2-dimethyl- piperazine-l-carboxylate (350 mg, 0.912 mmol, obtained in the preparation of Intermediate M) in DM F (8 mL) are added dicyanozinc (107 mg, 0.911 mmol) and Pd(PPh₃)₄ (211 mg, 0.182 mmol). The mixture is degassed for 5 min and then stirred at 100°C under nitrogen for 2h. Then the volatiles are removed under reduced pressure. The residue is purified on Biotage SNAP 25g silica gel cartridge eluting with EtOAc/hexane 0-30% in 20 CV to obtain tert-butyl 4-(6-cyano-3- methoxycarbonyl-2-pyridyl)-2,2-dimethyl-piperazine-l-carboxylate (280 mg, 82%) as a white solid. LC-MS(ESI): 375.33 (M+H)+; calc. 375.2032. RT: 1.86 min using Method C.

Step II: 6-cyano-2-(3,3-dimethylpiperazin-l-yl)pyridine-3-carboxylate (Dihydrochloride salt)

[0240] To a solution of tert-butyl 4-(6-cyano-3-methoxycarbonyl-2-pyridyl)-2,2-dimethyl-piperazine- 1-carboxylate (280 mg) in DCM (3 mL) is added HCI in dioxane (3.0 mL of 4 M, 12 mmol). The mixture is stirred at rt for 2h. Then the volatiles are removed under reduced pressure to provide methyl 6-cyano-2-(3,3-dimethylpiperazin-l-yl)pyridine-3-carboxylate (Dihydrochloride salt) (220 mg, 85%) as a white solid. LC-MS(ESI): 275.33 (M+H)+; calc. 275.1508. T: 0.64 min using Method C.

Preparation of Intermediate O. Methyl 6-(3,3-dimethyl-2-oxo-piperazin-l-yl)-2-methyl-pyridine-3- carboxylate (Dihydrochloride salt)

[0241] Step I: To a solution of tert-butyl 2,2-dimethyl-3-oxo-piperazine-l-carboxylate (1.60 g, 7.01 mmol) in DM F (10 mL) is added 60% NaH/mineral (280 mg, 7.00 mmol). The mixture is stirred at rt for 20 min, till the bubbling subsided. Then to it are added TBAI (70 mg, 0.19 mmol) and tert- butyl 2,2-dimethyl-3-oxo-piperazine-l-carboxylate (1.6 g, 7.1 mmol). The mixture is stirred at rt for lh under nitrogen, LC-MS of the crude shows a littlew conversion. Then the temperature is raised to 60°C, stirred for 3h. After cooling to rt, the mixture is neutralized with formic acid, diluted with EtOAc (80 mL), washed with water and brine consecutively, dried over sodium sulfate, filtered. The filtrate is concentrated to dryness under reduced pressure and the residue is purified on Biotage SNAP 50g silica gel cartridge eluting with EtOAc/hexanes 0-30% in 20CV to obtain tert-butyl 4-(5-methoxycarbonyl-6-methyl-2-pyridyl)-2,2-dimethyl-3-oxo-piperazine-l- carboxylate (755 mg, 37%) as a yellowish solid. LC-MS: 378.77 M(+H+), calc. 378.2028. RT: 1.9 min using Method C.

[0242] Step II: To a solution of tert-butyl 4-(5-methoxycarbonyl-6-methyl-2-pyridyl)-2,2-dimethyl-3- oxo-piperazine-l-carboxylate (755 mg) in DCM (2 mL) is added 4M HCI/dioxane (3 mL of 4 M, 12.00 mmol). The mixture is stirred at rt for 4 h. Then the mixture if filtered and the solid is dried in vacuo to provided methyl 6-(3,3-dimethyl-2-oxo-piperazin-l-yl)-2-methyl-pyridine-3- carboxylate (Dihydrochloride salt) (628 mg, 90%) as a white solid. LC-MS: 278.68 M(+H+), calc. 278.1504. RT: 0.56 min using Method C.

Preparation of Intermediate P. 6-chloro-2-methyl-pyridine-3-carbonitrile

[0243] Step I: (Z)-3-aminobut-2-enenitrile (2.87 g, 35 mmol) and ethyl prop-2-ynoate (3.6 mL, 35 mmol) are dissolved in dry DMF (17 mL). The reaction mixture is stirred for 1 h at room temperature, and the mixture is then heated at reflux (~160°C) for 3 days. The precipitate that forms after cooling to room temperature is collected by filtration under reduced pressure, washed with methanol (5 mL), ether (10 mL), and dried to afford 6-hydroxy-2-methyl-pyridine- 3-carbonitrile (438.3 mg, 8%) as a beige solid. ^XH NM (400 M Hz, DMSO-d6) δ 12.42 (brs, 1H), 7.59 (d, J = 9.5 Hz, 1H), 6.25 (d, J = 9.5 Hz, 1H), 2.38 (s, 3H). ESI-MS m/z calc. 134.04802, found 135.13 (M+l)+; Retention time: 0.13 minutes using Method J.

[0244] Step II: A 10 mL round bottom flask is charged with 6-hydroxy-2-methyl-pyridine-3- carbonitrile (436 mg, 3.25 mmol) and POCI₃ (910 μί, 9.76 mmol) is added and a reflux condenser attached. The slurry is heated at 100°C for 2 hours. The mixture is cooled down to room temperature and then to 0°C before a saturated NaHC0₃ aqueous solution (about 10 mL) is added very carefully. The mixture is diluted with DCM (15 mL) and carefully poured in more saturated NaHC0₃ aqueous solution (50 mL). The phases are separated and the aqueous layer is extracted twice with DCM. The combined organic layers are dried over MgS0₄, filtered and concentrated under reduced pressure. The crude product is purified by flash chromatography on silica gel eluting with hexanes and EtOAc to afford the desired 6-chloro-2-methyl-pyridine-3- carbonitrile (404 mg, 76%) as a white solid. ^XH NM R (400 MHz, Chloroform-d) δ 7.83 (d, J = 8.2 Hz, 1H), 7.30 (d, J = 8.2 Hz, 1H), 2.77 (s, 3H). ESI-MS m/z calc. 152.01413, found 153.16 (M+l)+; Retention time: 0.52 minutes using Method J.

Preparation of Intermediate Q. Ehyl 5-(3,3-dimethylpiperazin-l-yl)imidazo[l,2-alpyridine-2- carboxylate (Dihydrochloride salt)

[0245] Step I: To a solution of tert-butyl 2,2-dimethylpiperazine-l-carboxylate (400 mg, 1.87 mmol) in DMF (3 mL) are added ethyl 5-bromoimidazo[l,2-a]pyridine-2-carboxylate (502 mg, 1.87 mmol), K₂C0₃ (516 mg, 3.73 mmol), X-PHOS (178 mg, 0.373 mmol) and Pd₂(dba)₃ (171 mg, 0.187 mmol) in DMF (3 mL). After degassing for 5 min, the mixture is stirred at 90°C under nitrogen overnight. Then the mixture is diluted with EtOAc (60 mL), washed with water and brine, dried over sodium sulfate and, filtered. The filtrate is concentrated to dryness under reduced pressure. The residue is purified on Biotage SNAP 25g silica gel cartridge eluting with EtOAc/hexanes 0- 50% in 20 CV to obtain a yellowish solid, mainly containing ethyl 5-(4-tert-butoxycarbonyl-3,3- dimethyl-piperazin-l-yl)imidazo[l,2-a]pyridine-2-carboxylate (250 mg, 33%), which is used directly in the next step without further purification. LC-M E(ESI): 403.25 (M+H)+; calc. 403.2345. RT: 1.78 min using Method C.

[0246] Step II: To the mixture from step 1 in DCM (1 mL) is added 4M HCI/dioxane (500 μΐ of 4 M, 2.00 mmol). The mixture is stirred at rt for 2h. Then it is filtered and the solid is washed with DCM, dried in vacuo to provide ethyl 5-(3,3-dimethylpiperazin-l-yl)imidazo[l,2-a]pyridine-2- carboxylate (Dihydrochloride salt) (120 mg, 52%) as an off-white solid. LVC-MS (ESI): 303.20 (M+H)+; calc. 303.1821. RT: 0.68 min using Method C.

Preparation of Intermediate R. (3R)-4-methyl-3-(2H-tetrazol-5-yl)pentanoic acid

[0247] Step I: To a solution of (2R)-4-tert-butoxy-2-isopropyl-4-oxo-butanoic acid (700 mg, 3.24 mmol) in THF (10.0 mL) is added CDI (577 mg, 3.56 mmol) at room temperature. After being stirred for 2.5 hours, NH₄OH solution (10.5 mL of 28 %w/w, 75.3 mmol) is added slowly and the mixture stirred for 90 minutes before being partitioned between water (40 mL) and EtOAc (25 mL). The phases are separated and the aqueous layer is extracted twice with EtOAc (2 X 25 mL). The combined organic layers are dried over MgS0₄, filtered and concentrated under reduced pressure to afford tert-butyl (3R)-3-carbamoyl-4-methyl-pentanoate (695 mg, 100%) as a colorless residue. The product isused for the next step without further purification. ESI-MS m/z calc. 215.15215, found 216.2 (M+l)+; Retention time: 0.54 minutes using Method J.

[0248] Step II: Trifluoroacetic anhydride (550 μί, 3.96 mmol) is added dropwise to a cooled solution of tert-butyl (3R)-3-carbamoyl-4-methyl-pentanoate (813 mg, 3.78 mmol) and pyridine (610 μί, 7.54 mmol) in DCM (15 mL) at 0°C. The reaction mixture is stirred at 0°C for 1 hour and is treated with a saturated NaHC03 aqueous solution (5 mL). Then, water (30 mL) and DCM (10 mL) are added. The phases are separated and the aqueous layer is extracted twice with DCM. The combined organic layers are dried over MgS0₄, filtered and concentrated under reduced pressure to afford the title compound along with pyridine according to proton NM analysis. The residue is partitionned between DCM and a saturated NH₄CI aqueous solution. The phases are separated and the aqueous layer is extracted twice with DCM. The combined organic layers are dried over MgS0₄, filtered and concentrated to dryness under high vacuum for 1 hour to afford pure tert-butyl (3R)-3-cyano-4-methyl-pentanoate (510 mg, 68%) as a yellow residue. ¹H NMR (400 M Hz, Chloroform-d) δ 2.92 (ddd, J = 8.6, 6.1, 5.1 Hz, 1H), 2.59 (dd, J = 16.3, 8.7 Hz, 1H), 2.46 (dd, J = 16.3, 6.2 Hz, 1H), 1.97 - 1.80 (m, 1H), 1.47 (s, 9H), 1.07 (dd, J = 10.2, 6.7 Hz, 6H). ESI-MS m/z calc. 197.14159, found 198.16 (M+l)+; Retention time: 0.83 minutes using Method J (mass detection only).

[0249] Step III: NaN₃ (24.0 mg, 0.369 mmol) and Ν,Ν-diethylethanamine (Hydrochloride salt) (50.0 mg, 0.363 mmol) are added to a solution of tert-butyl (3R)-3-cyano-4-methyl-pentanoate (22.0 mg, 0.112 mmol) in DM F (500 μί). The reaction mixture is exposed to microwave irradiation at 150 °C for 2 h. Further NaN₃ (54.0 mg, 0.831 mmol) and Ν,Ν-diethylethanamine (Hydrochloride salt) (117 mg, 0.850 mmol) are added and the mixture is exposed again to microwave irradiation at 150 °C for 2h. Then, water (0.5 mL) is added and the mixture is forozen and is lyophilised over the weekend. The obtained beige residue is suspended in dioxane and the mixture is filtered under reduced pressure. The resulting filtrate is used as is for the next step. ESI-MS m/z calc. 240.15863, found 239.09 (M-l)-; Retention time: 0.56 minutes using Method J (mass detection only).

[0250] Step IV: HCI in dioxane (500 μΐ of 4 M, 2.00 mmol) is added to a solution of tert-butyl (3R)- 4-methyl-3-(2H-tetrazol-5-yl)pentanoate (27.0 mg, 0.112 mmol) in dioxane (1.5 mL) at room temperature. The mixture is heated at 50°C in an oil bath and stirred for 18 hours. The volatiles are removed under reduced pressure to afford (3R)-4-methyl-3-(2H-tetrazol-5-yl)pentanoic acid as a dark yellow residue. The product isused as such without further purification. ESI-MS m/z calc. 184.09602, found 183.15 (M-l)-; Retention time: 0.25 minutes using Method J (mass detection only).

Preparation of Intermediate S. 2-(2-methoxycarbonyltetrahydrofuran-2-yl)acetic acid

[0251] Step I: A solution of diisopropylamine (6.0 mL, 43 mmol) in THF (100 mL) is cooled to -78°C and treated with butyllithium (19.2 mL of 2.5 M, 48.0 mmol). After stirring for 30 min, the mixture is treated with methyl tetrahydrofuran-2-carboxylate (5.00 g, 38.4 mmol) and the solution stirred for 45 min. The mixture is then treated with tert-butyl 2-bromoacetate (15.0 g, 76.9 mmol) and allowed to warm to RT overnight. The mixture is treated with sat NH₄CI and the layers are separated. The aq layer is extracted with EtOAc (X3). The combined organic layers are washed (H₂0, brine), dried (Na₂S0₄) and concentrated in vacuo. The residue is purified by column chromatography (silica gel, Isco Combiflash, 0 to 100% EtOAc in hexanes). Note that 3 purifications are required to achieve good purity of the mateiral. Obtained a light yellow oil (2.159 g, 23%). ^XH NM R (400 MHz, Chloroform-d) δ 3.99 (td, J = 6.3, 3.9 Hz, 2H), 3.76 (d, J = 0.7 Hz, 3H), 2.90 (d, J = 15.5 Hz, 1H), 2.66 (d, J = 15.5 Hz, 1H), 2.29 (hept, J = 6.6 Hz, 1H), 1.95 (tdd, J = 9.7, 8.3, 4.5 Hz, 3H), 1.43 (d, J = 0.6 Hz, 9H).

[0252] Step II: A solution of methyl 2-(2-tert-butoxy-2-oxo-ethyl)tetrahydrofuran-2-carboxylate (1.00 g, 4.09 mmol) in DCM (5 mL) and TFA (5.0 mL, 65 mmol) is allowed to stir at rt for 4h. The solvent is removed in vacuo and the crude material is used without further purification. Obtained a colorless oil (701 mg, 91%). ^XH NMR (400 M Hz, Chloroform-d) δ 4.15 - 4.01 (m, 2H), 3.78 (s, 3H), 3.12 (d, J = 16.2 Hz, 1H), 2.79 (d, J = 16.2 Hz, 1H), 2.30 (dt, J = 12.7, 6.6 Hz, 1H), 2.12 - 1.84 (m, 3H).

Preparation of Intermediate T. methyl 6-(3,3-dimethylpiperazin-l-yl)-3-methyl-pyridine-2- carboxylate (hydrochloride salt)

T

[0253] Step I: A mixture of 6-chloro-3-methyl-pyridine-2-carbonitrile (914 mg, 5.99 mmol), 2,2- dimethylpiperazine (1000 μί, 7.30 mmol) and cesium carbonate (2.27 g, 6.97 mmol) in NMP (12 mL) is heated at 100°C overnight. The reaction is cooled down to room temperature and water (120 mL) is added. The mixture is extracted with Et₂0 (3 x 50 mL). The combined organic phases are washed with water (60 mL) and brine (60 mL), dried over Na₂S0₄ and concentrated under reduced pressure. The crude material isused without further purification in the next step (1.082 g, 69%). ESI-MS m/z calc. 230.15315, found 230.68 (M+l)+; Retention time: 1.03 minutes using Method L.

[0254] Step II: 6-(3,3-dimethylpiperazin-l-yl)-3-methyl-pyridine-2-carbonitrile (1.08 g, 4.70 mmol) is added to potassium hydroxide (8.0 mL of 6 M, 48 mmol) and the mixture is stirred at 100°C for 3h. The reaction mixture is acidified with HCI (3 M) and the mixture is concentrated to dryness under reduced pressure. The solid residue is dissolved in dry MeOH (23.5 mL). H₂S0₄ (1.00 mL, 18.7 mmol) is added and the mixture is stirred at reflux overnight. The mixture is neutralized with a saturated solution of K₂C0₃ and extraced with EtOAc (2 x 20 mL). The combined organic phases are washed with brine, dried over Na₂S0₄ and concentrated under reduced pressure. The residue ispurified by reverse phase chromatography (0 % to 100 % ACN in water) yielding methyl 6-(3,3-dimethylpiperazin-l-yl)-3-methyl-pyridine-2-carboxylate (298 mg, 22%) 1H NMR (400 MHz, DMSO-d6) δ 7.42 (dd, J = 8.7, 0.7 Hz, 1H), 6.90 (d, J = 8.7 Hz, 1H), 3.78 (s, 3H), 3.44 - 3.33 (m, 2H), 3.18 (s, 2H), 2.88 - 2.73 (m, 2H), 2.21 (s, 3H), 1.04 (s, 6H). ESI-MS m/z calc. 263.1634, found 264.57 (M+l)+; Retention time: 1.16 minutes using Method L.

Preparation of Intermediate U. Methyl 2-amino-6-(3,3-dimethylpiperazin-l-yl)pyridine-3-carboxylate

(Dihydrochloride salt)

[0255] Step I: To a solution of tert-butyl 2,2-dimethylpiperazine-l-carboxylate (6.318 g, 29.48 mmol) in NMP (20 mL) are added methyl 2-amino-6-chloro-pyridine-3-carboxylate (5.00 g, 26.8 mmol) and DIPEA (9.34 mL, 53.6 mmol). The mixture is heated in a microwave at 160°C for 3h. After cooling to rt, the mixture is diluted with water, stirred for lh, then it is filtered off. The solid is washed with water, dried in vacuo, purified on Biotage SNAP 100 g silica gel cartridge eluting with EtOAc/hexanes 0-25% in 20 CV to obtain tert-butyl 4-(6-amino-5-methoxycarbonyl- 2-pyridyl)-2,2-dimethyl-piperazine-l-carboxylate (4.61 g, 47%) as a white solid. ESI-LC-MS: 365.42 (M+H)+; calc. 365.2188. RT: 1.74 min using Method C. [0256] Step II: To a solution of tert-butyl 4-(6-amino-5-methoxycarbonyl-2-pyridyl)-2,2-dimethyl- piperazine-l-carboxylate (4.61 g) in DCM (10 mL) is added 4M HCI/dioxane (15.0 mL of 4 M, 60.0 mmol). The mixture is stirred at rt overnight. Then it is diluted with DCM. The white suspension is filtered and the solid is washed with DCM, dried in vacuo to afford methyl 2-amino-6-(3,3- dimethylpiperazin-l-yl)pyridine-3-carboxylate (Dihydrochloride salt) (4.2 g, 99%) as a white solid. ESI-LC-MS: 265.36 (M+H)+, calc. 265.1664. RT. 0.74 min using Method C.

Preparation of Intermediate W. (4-amino-2,2-dimethyl-l-piperidyl)-[7-tert-butyl-5-(4-chloro-3- fluoro- phenyl)furo[3,2-blpyridin-2-yllmethanone

[0257] Step I: DIPEA (1.364 g, 1.838 mL, 10.55 mmol) is added to a stirred mixture of 7-tert-butyl-5- (4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carboxylic acid (815 mg, 2.34 mmol) and HATU (1.337 g, 3.516 mmol) in DMF (8.0 mL). 2,2-dimethylpiperidin-4-one (Hydrochloride salt) (423.6 mg, 2.459 mmol) is then added and the reaction is stirred at rt for overnight. Water is added along with EtOAc and the phases are separated. The organic phase is washed 2 other times with water and brine (1:1 mixture), dried over MgS04, filtered and evaporated under reduced pressure. The recovered crude compound is purified on Isco Combiflash 40 g silica gel cartridge eluting with 0-100% EtOAc/Hexanes (loading with DCM) to give l-[7-tert-butyl-5-(4-chloro-3- fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-2,2-dimethyl-piperidin-4-one (785 mg, 73%) ESI- MS m/z calc. 456.1616, found 459.15 (M+l)+; Retention time: 1.05 minutes using Method J.

[0258] Step II: A mixture of l-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2- carbonyl]- 2,2-dimethyl-piperidin-4-one (350 mg, 0.766 mmol), Titanium(IV) isopropoxide (435.4 mg, 452.1 μί, 1.53 mmol) and Ammonia in MeOH (547.1 μΐ of 7 M, 3.830 mmol) are stirred under argon in a capped flask at rt for 6.5h. NaBH₄ (43.5 mg, 1.15 mmol) is added and the reaction is stirred at r.t. for 2 days. The reaction is quenched by pouring into ammonium hydroxide 10% aqueous solution. DCM is added and the organic phase is separated. The aqueous phase is washed with DCM and combined filtrates are concentrated to give upon trituration crude (4-amino-2,2-dimethyl-l-piperidyl)-[7-tert-butyl-5- (4-chloro-3-fluoro- phenyl)furo[3,2-b]pyridin-2-yl]methanone (Formate (0.5)) (307 mg, 81%) ESI-MS m/z calc. 457.19324, found 456.15 (M+l)+; Retention time: 2.72 minutes using Method A. The material is further purified by UV directed reverse phase HPLC to give (4-amino-2,2-dimethyl-l-piperidyl)- [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]methanone (95 mg) as a white solid. ^XH NM R (400 M Hz, DMSO-d6) δ 8.17 (dd, J = 11.0, 2.1 Hz, 1H), 8.07 - 8.00 (m, 1H), 7.84 (s, 1H), 7.77 - 7.68 (m, 1H), 7.53 (s, 1H), 7.26 (s, 2H, br), 3.90 - 3.79 (m, 1H), 3.47 - 3.17 (m, 2H), 2.12 - 1.98 (m, 1H), 1.87 - 1.77 (m, 1H), 1.71 - 1.49 (m, 14H), 1.45 (s, 3H). ESI-MS m/z calc. 457.19324, found 456.15 (M+l)+; Retention time: 2.72 minutes using method A.

Preparation of Intermediate X. [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridin-2-yll-

[2,2-dimethyl-4-(methylamino)-l-piperidyllmethanone

[0259] To a 1,2-dichloroethane (3.1 mL) solution of l-[7-tert-butyl-5-(4-chloro-3-fluoro- phenyl)furo[3,2-b]pyridine-2-carbonyl]-2,2-dimethyl-piperidin-4-one (350 mg, 0.766 mmol), MeNH₂ (144 μΐ of 33 %w/v, 1.53 mmol), Sodium Triacetoxyborohydride (121.8 mg, 0.5745 mmol) and Acetic Acid (50 μί, 0.88 mmol) are added. The mixture is stirred at rt for 48 hours. Sodium triacetoxyborohydride (81.17 mg, 0.3830 mmol) and MeNH₂ (80 μΐ of 33 %w/w) are added. The reaction is stirred for 2 hours at rt. Water is added along with DCM and the phases are separated. The organic phase is washed with water and brine, dried over MgS0₄, filtered and evaporated under reduced pressure. The residue is dissolved in EtOAc and HCI (210.6 μί οί 4 M, 0.843 mmol) in dioxane is added. The solution is concentrated. The residue is diluted in NM P and purified on a Gemini-NX 250-21 column (Isco Combiflash EZ PREP) eluting with 40-70% CH3CN (0.1% Formic Acid)/Water (0.1% Formic Acid) over 30 minutes. Desired fractions are combined and lyophilized to give [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2- yl]-[2,2-dimethyl-4-(methylamino)-l-piperidyl]methanone (145 mg, 40%) ESI-MS m/z calc. 471.2089, found 472.25 (M+l)+; Retention time: 2.7 minutes using Method A.

Preparation of Intermediate Y. (4-amino-2,2-dimethylpiperidin-l-yl)(7-(tert-butyl)-5-(4,4- dimethylcvclohexyl)furo[3,2-blpyridin-2-yl)methanone

[0260] Step I: DIPEA (1.96 g, 2.64 mL, 15.2 mmol) is added to a stirred mixture of 7-tert-butyl-5- (4,4-dimethylcyclohexyl)furo[3,2-b]pyridine-2-carboxylic acid (1.00 g, 3.04 mmol) and HATU (1.50 g, 3.95 mmol) in DM F (9.8 mL). To this mixture is added 2,2-dimethylpiperidin-4-one (Hydrochloride salt) (548.6 mg, 3.185 mmol) and the reaction is stirred at rt for overweekend. Water is added along with EtOAc and the phases are separated. The organic phase is washed 2 other times with water and brine (1:1 mixture), dried over MgS04, filtered and evaporated under reduced pressure. The crude compound is purified on Isco Combiflash 80 g silica gel cartridge eluting with 0-100% EtOAc/Hexanes (loading with DCM). Selected fractions are combined and concentrated. The residue is triturated in EtOAc/Heptane to give l-[7-tert-butyl- 5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridine-2-carbonyl]-2,2-dimethyl-piperidin-4-one (1.025 g, 77%) an off-white solid. ESI-MS m/z calc. 438.28824, found 439.36 (M+l)⁺; Retention time: 1.11 minutes using Method J.

[0261] Step II: To a stirred solution of l-[7-tert-butyl-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridine- 2-carbonyl]-2,2-dimethyl-piperidin-4-one (260 mg, 0.5928 mmol) in Ethanol (1.0 mL) is added Ammonium formate (186.9 mg, 2.964 mmol). After 5 minutes of stirring, Pd-C (315.4 mg of 20 %w/w, 0.593 mmol) is added. The reaction mixture is stirred at rt overnight. The reaction mixture is filtered on a pre-packed celite pad (wash with EtOH and DCM). The filtrate is concentrated and the crude residue is dissolved in NM P and purified on a Gemini-NX 250-21 column (Isco Combiflash EZ PREP) eluting with 35-65% CH3CN (0.1% Formic Acid)/Water (0.1% Formic Acid) on a 30 minutes run. Desired fractions are combined and lyophilized to give (4- amino-2,2-dimethyl-l-piperidyl)-[7-tert-butyl-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridin-2- yljmethanone (85 mg, 33%) as a white solid. ESI-MS m/z calc. 439.3199, found 440.84 (M+l)+; Retention time: 1.61 minutes using Method C.

Preparation of Intermediate Z. (7-(tert-butyl)-5-(4,4-dimethylcvclohexyl)furo[3,2-blpyridin-2-yl)(2,2- dimethyl-4-(methylamino)piperidin-l-yl)methanone

[0262] Intermediate Z was prepared analogously to Intermediate X, beginning with Intermediate C. Preparation of Intermediate AA: tert-butyl 6-(3,3-dimethylpiperazin-l-yl)pyridine-2-carboxylate

AA

[0263] The intermediate is prepared according to General Procedure 22A using a solution of tert- butyl 6-fluoropyridine-2-carboxylate (30.05 g, 152.4 mmol), 2,2-dimethylpiperazine (21.77 g, 190.6 mmol) and cesium carbonate (61.02 g, 187.3 mmol) in NMP (300 mL) at 100°C overnight.

Preparation of Intermediate AB. 6-(4-(7-(tert-butyl)-5-(4,4-dimethylcvclohexyl)furo[3,2-blpyridine-2- carbonyl)-3,3-dimethylpiperazin-l-yl)picolinic acid

[0264] The product is prepared according to General Procedure 1 using Intermediates C and AA in NM P (7.2 mL) using DIPEA (636 μΐ, 3.65 mmol) and HATU (716 mg, 1.88 mmol) to afford the tert-butyl intermediate (770 mg, 1.28 mmol). The ester intermediate is then hydrolyzed using General Procedure 23, in 1,4-dioxane (7.7 mL) with HCI (1.6 mL of 4 M, 6.4 mmol) in dioxane to generate 6-[4-[7-tert-butyl-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-piperazin-l-yl]pyridine-2-carboxylic acid. ESI-MS m/z calc. 546.3206, found 547.02 (M+l)+;

Preparation of Intermediate AC. tert-butyl 6-(3,3-dimethylpiperazin-l-yl)pyridine-2-carboxylate

AC

[0265] Step I: A mixture of tert-butyl 2,2-dimethylpiperazine-l-carboxylate (838 mg, 3.91 mmol), ethyl 6-bromopyridine-2-carboxylate (989 mg, 4.30 mmol), Cs₂C0₃ (3.18 g, 9.77 mmol), in NM P (8.38 mL) is heated overnight at 120°C. The reaction mixture is diluted with DCM (2mL) and purified on silica column eluting with 10-40% ethylacetate:hexanes to afford the N-Boc intermediate (254 mg, 18%).

[0266] Step II: To tert-butyl 4-(6-ethoxycarbonyl-2-pyridyl)-2,2-dimethyl-piperazine-l-carboxylate in dioxane (8.4 mL) and MeOH (837.8 μί) is then added HCI (977 μΐ of 4 M, 3.91 mmol) and the reaction mixture stirred at rt overnight. The solvent is evaporated in vacuo to give the crude title product.

Preparation of Intermediate AD. 6-(4-(7-(tert-butyl)-5-(4-chlorophenyl)furo[3,2-blpyridine-2- carbonyl)-3,3-dimethylpiperazin-l-yl)picolinic acid

[0267] The ethyl ester is prepared according to General Procedure 1 using Intermediates B and AC, DIPEA (1.08 g, 1.45 mL, 8.35 mmol) and HATU (635 mg, 1.67 mmol) in DM F (4.4 mL) for 1 h at r.t. The ethyl ester is then hydrolyzed according to General Procedure 11 using LiOH (4 eq, 2.5 mmol, 1.2 mL, 2M) in dioxane (4.4 mL) for 4 h at r.t. to afford the title product (347 mg, 36%) ^XH NM (400 MHz, dmso) δ 12.57 (s, 1H), 8.19 - 8.07 (m, 2H), 7.75 (s, 1H), 7.69 (dd, J = 8.6, 7.2 Hz, 1H), 7.62 - 7.51 (m, 3H), 7.27 (d, J = 7.2 Hz, 1H), 6.84 (d, J = 8.6 Hz, 1H), 4.12 - 4.03 (m, 2H), 3.98

- Ill - (s, 2H), 3.66 - 3.60 (m, 2H), 1.61 - 1.40 (m, 15H). ESI-MS m/z calc. 546.20337, found 547.48 (M+l)+.

Preparation of Intermediate AE. 2-[4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2-blpyridine-2- carbonyll-3,3-dimethyl-piperazin-l-yllpyrimidine-5-carboxylic acid

[0268] Step I: Three microwave vials are each charged with one-third of the following: [7-tert- butyl-5-(4-chlorophenyl)furo[3,2-b]pyridin-2-yl]-(2,2-dimethylpiperazin-l-yl)methanone

(Dihydrochloride salt) (3.02 g, 6.05 mmol) and methyl 2-chloropyrimidine-5-carboxylate (1.57 g, 9.07 mmol). To each tube is added one-third of the following: dioxane (48 mL) and DIPEA (5.4 mL, 31.0 mmol) and the tubes are capped and submitted in queue to microwave for 10 min at 120 °C. The contents of the tubes are combined and the solvent is removed under reduced pressure. The residue is loaded on silica using DCM/MeOH and purified on Biotage SNAP 100 g column. Eluent is EtOAc in hexanes, 0% (1CV), 0-40% (12CV), 40% (2CV). Methyl 2-[4-[7-tert- butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-l-yl]pyrimidine- 5-carboxylate (3.02 g, 89%) is obtained as a light yellow solid. ¹H NM R (400 MHz, DMSO) δ 8.84 (d, J = 7.0 Hz, 2H), 8.18 - 8.06 (m, 2H), 7.77 (s, 1H), 7.58 - 7.54 (m, 2H), 7.54 (s, 1H), 4.20 - 4.00 (m, 4H), 3.81 (m, 5H), 1.54 (s, 6H), 1.53 (s, 9H). Contains 2.36% wt DCM, traces of solvents (EtOAc, hexanes). ESI-MS m/z calc. 561.2143, found 564.15 (M+l)+; Retention time: 1.19 minutes using Method J.

Step II: 2-[4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl- piperazin-l-yl]pyrimidine-5-carboxylic acid [0269] To a suspension of methyl 2-[4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2- carbonyl]-3,3-dimethyl-piperazin-l-yl]pyrimidine-5-carboxylate (3.02 g, 5.37 mmol) in MeOH (39 mL) is added NaOH, (5.4 mL of 2 M, 10.8 mmol). After stirring for 90 min, THF (26 mL) is added followed by further NaOH, aqueous (2.7 mL of 2 M, 5.40 mmol) after 3h. The reaction is allowed to stir for a further 36 h. The reaction mixture is concentrated to ca. 1/2 the initial volume in vacuo and treated with H₂0 (30 mL). The reaction mixture is acidified to pH 4 with 2 N HCI (ca. 8.5 mL). The mixture is allowed to stir until a solid is obtained and the material is collected by filtration and washed with H₂0. The solid is air dried followed by drying overnight in a vacuum oven at 45°C. Obtained 2.7934g of white solid. The material is taken up in DCM (60 mL, 21.5 vol eq) and EtOH (18 mL,6.5 vol eq).With brief heating. The solution is treated with 1.70g Silia MetS SiDMT (0.59 mmol/g), stirred overnight. The mixture is filtered, the solid rinsed with 20% EtOH in DCM and the solvents removed in vacuo. The residue is suspended in H₂0 (40 mL) and allowed to stir for 4h then filtered and allowed to dry for 48 h under suction with in inverted funnel and flow of N₂ (gas). The material is then dried in a vacuum oven overnight to afford 2-[4-[7-tert- butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-l-yl]pyrimidine- 5-carboxylic acid (2.53 g, 85%) is obtained as a white solid. ^XH NM (400 MHz, DMSO) δ 12.85 (s, 1H), 8.81 (s, 2H), 8.15 (d, J = 8.6 Hz, 2H), 7.77 (s, 1H), 7.56 (d, J = 8.6 Hz, 2H), 7.54 (s, 1H), 4.13 - 4.04 (m, 4H), 3.84 - 3.74 (m, 2H), 1.53 (2s, 15H). ESI-MS m/z calc. 547.1986, found 549.56 (M+l)+; Retention time: 4.33 minutes using Method A.

Preparation of Intermediate AF. 2-(4-(7-(tert-butyl)-5-(4-chloro-3-fluorophenyl)furo[3,2-blpyridine- 2-carbonyl)-3,3-dimethylpiperazin-l-yl)pyrimidine-5-carboxylic acid

[0270] Intermediate AF is prepared analogously to Intermediate AE beginning with Intermediate D.

Preparation of Intermediate AG: l-[l-[(4-methoxyphenyl)methyllpyrazol-4-yll-3,3-dimethyl- piperazin-2-one (Hydrochloride salt)

AG

Step I: 4-iodo-l-[(4-methoxyphenyl)methyl]pyrazole [0271] A mixture of 4-iodo-lH-pyrazole (26 g, 134 mmol), l-(chloromethyl)-4-methoxy-benzene (30.1 g, 192.4 mmol), cesium carbonate (64.7 g, 198.6 mmol) in DMF (267 mL) is heated at 60°C overnight. The reaction mixture is diluted with 250 mL water and extracted with 2 X 250 mL ethyl acetate. The combined organic layers are washed with brine then dried over Na₂S0₄. The mixture is filtered then evaporated and the residue pre-adsorbed on silica gel. Purified by silica gel plug filtration using 0-10% EtOAc/Hexanes as eluent.

Step II: l-[l-[(4-methoxyphenyl)methyl]pyrazol-4-yl]-3,3-dimethyl-piperazin-2-one

[0272] A mixture of tert-butyl 2,2-dimethyl-3-oxo-piperazine-l-carboxylate (3.42 g, 15.0 mmol), 4- iodo-l-[(4-methoxyphenyl)methyl]pyrazole (5.18 g, 16.5 mmol), iodocopper (1.43 g, 7.50 mmol), N,N'-dimethylethane-l,2-diamine (1.32 g, 1.60 mL, 15.0 mmol), K₃P0₄ (6.37 g, 30.0 mmol) in DM F (78.8 mL) is heated at 120°C for 4h. The reaction mixture is diluted with water (150mL) and extracted (2X) with EtOAc (2X150mL). The combined organics are washed with waer (150mL) and brine (100m L), evaporated in vacuo and purified by flash column chromatography.

Step III: l-[l-[(4-methoxyphenyl)methyl]pyrazol-4-yl]-3,3-dimethyl-piperazin-2-one (Hydrochloride salt)

[0273] To the material obtained above, in dioxane (34.2 mL) and MeOH (1.7 mL) is added HCI (18.8 mL, 4 M, 75. 0 mmol). The reaction is allowed to stir at T for 4h. The solvent is removed in vacuo to give a gummy solid. This is triturated with DCM (200mL) to give a white suspension which is suction-filtered to give a white powder. l-[l-[(4-methoxyphenyl)methyl]pyrazol-4-yl]- 3,3-dimethyl-piperazin-2-one (Hydrochloride salt) (5.41 g, 15.42 mmol, 103%) ^XH NMR (400 MHz, DMSO-d6) δ 10.18 (s, 2H), 8.13 (d, J = 0.8 Hz, 1H), 7.66 (d, J = 0.8 Hz, 1H), 7.24 - 7.11 (m, 2H), 6.93 - 6.75 (m, 2H), 5.19 (s, 2H), 3.93 - 3.83 (m, 2H), 3.69 (s, 3H), 3.58 - 3.45 (m, 2H), 1.55 (s, 6H).

Preparation of Intermediate AH: 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2- carbonyll-3,3-dimethyl-l-(lH-pyrazol-4-yl)piperazin-2-one

Step I: 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-l-[l-[(4- methoxyphenyl)methyl]pyrazol-4-yl]-3,3-dimethyl-piperazin-2-one

[0274] To a solution of 7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carboxylic acid (600 mg, 1.73 mmol) (Intermediate A) in DMF (4 mL) are added l-[l-[(4- methoxyphenyl)methyl]pyrazol-4-yl]-3,3-dimethyl-piperazin-2-one (Hydrochloride salt) (605 mg, 1.72 mmol) (Intermediate AG), [dimethylamino(triazolo[4,5-b]pyridin-3-yloxy)methylene]- dimethyl-ammonium;hexafluorophosphate (700 mg, 1.84 mmol) and DIPEA (1.5 mL, 8.612 mmol). The mixture is stirred at r.t. for 24 h. Then it is diluted with water, stirred at rt for 1 h, and filtered. The solid is washed with water, dried under reduced pressure. The residue is purified by flash column chromatography eluting with EtOAc/hexanes 0-70% to obtain 4-[7-tert- butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-l-[l-[(4- methoxyphenyl)methyl]pyrazol-4-yl]-3,3-dimethyl-piperazin-2-one (592 mg, 53%) as a white solid. LC-MS(ESI): 644.48 (M+H)+; calc. 644.2439. RT: 2.2 min, Method C.

Step II: 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-l-(lH-pyrazol-4-yl)piperazin-2-one

[0275] A solution of 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-l- [l-[(4-methoxyphenyl)methyl]pyrazol-4-yl]-3,3-dimethyl-piperazin-2-one (592 mg) in TFA (2 mL) is heated at 130 °C for lh in microwave. The volatiles are removed under reduced pressure. The residue is dissolved in MeOH, diluted with water and neutralized with NaHC0₃ solution, filtered. The solid is washed with water, dried in vacuo. The residue is purified by flash column chromatography eluting with EtOAc/DCM 50-100% to obtain 4-[7-tert-butyl-5-(4-chloro-3- fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-l-(lH-pyrazol-4-yl)piperazin-2-one (370 mg, 79%) as a white solid. LC-ME(ESI): 524.36 (M+H)+; calc. 524.1864. RT: 1.85 min, Method C.

Preparation of Intermediate Al. c/^'5-4-(4-(7-(tert-butyl)-5-(4,4-dimethylcvclohexyl)furo[3,2- blpyridine-2-carbonyl)-3,3-dimethylpiperazine-l-carbonyl)-l-methylcvclohexane-l-carboxylic acid

Step I: 4-methoxycarbonyl-4-methyl-cyclohexanecarboxylic acid

[0276] To a solution of LDA (in THF/heptanes/ethylbenzene) (33.6 mL of 2 M, 67.1 mmol) in THF (30 mL) at 40°C is added a solution of 4-methoxycarbonylcyclohexanecarboxylic acid (5.00 g, 26.9 mmol) in THF 50 mL over 5 minutes keeping temperature below -20°C. The reaction mixture is allowed to warm slowly to 0°C (ice bath). To this mixture is added Mel (2.51 mL, 40.3 mmol) slowly keeping temperature below 20°C. After stirring for ca. 2h the mixture is re-cooled in in an ice bath, and treated with 50 mL of HCI 2N dropwise (pH = 6-7) then warmed to room temperature. The mixture is treated with 10 mL of HCI 2N (pH = 1) extracted with EtOAc (3X100 mL), the combined organic layers are washed with 25 mL of brine, dried over Na₂S0₄, and concentrated under vacuum. The residue is recrystallized from iPrOAc to give the desired acid (23% recovery). This material is used as is in subsequent experiments.

Step II: l-methylcyclohexane-l,4-dicarboxylic acid

[0277] A mixture of 4-methoxycarbonyl-4-methyl-cyclohexanecarboxylic acid (449.8 mg, 2.25 mmol), NaOH (2.25 mL of 2 M, 4.49 mmol) in dioxane (8.1 mL) is heated at 80°C for 4.5h. The solvent is evaporated in vacuo and the reaction cooled to 0°C. HCI (2.8 mL of 2 M, 5.6 mmol) is added and the resultant solid is suction filtered to give a white solid l-methylcyclohexane-1,4- dicarboxylic acid (393 mg, 94%). ^XH NM (400 M Hz, dmso) δ 12.08 (s, 1H), 2.16 - 2.04 (m, OH), 2.04 - 1.96 (m, 1H), 1.71 (dt, J = 13.8, 3.7 Hz, 1H), 1.30 (qd, J = 13.2, 3.1 Hz, 1H), 1.13 - 0.94 (m, 2H).

Step III: [0278] To a mixture of [7-tert-butyl-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridin-2-yl]-(2,2- dimethylpiperazin-l-yl)methanone (Hydrochloride salt) (Intermediate E) (60.0 mg, 0.130 mmol) and l-methylcyclohexane-l,4-dicarboxylic acid (36.3 mg, 0.195 mmol) in DMF (589 μί) is added DIPEA (136 μί, 0.779 mmol) and T₃P (99.2 μΐ of 50 %w/v in DMF, 0.156 mmol) dropwise. The reaction is stirred for 1.5 h, after which LCMS indicated full conversion. The reaction mixture is diluted with water (5 mL), 1M citric acid (5 mL), and extracted twice with ethyl acetate (10 mL). The solvent is removed under reduced pressure. The residue is purified by reverse phase preparative HPLC to give the desired product after lyophilisation: 4-[4-[7-tert-butyl-5-(4,4- dimethylcyclohexyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazine-l-carbonyl]-l- methyl-cyclohexanecarboxylic acid (29 mg, 37%). ^XH NM (400 M Hz, DMSO-d6) δ 12.21(brs, 1H), 7.38 (d, J = 17.0 Hz, 1H), 7.12 (d, J = 3.8 Hz, 1H), 3.92 - 3.80 (m, 2H), 3.73 (d, J = 5.4 Hz, 1H), 3.62 (d, J = 26.4 Hz, 2H), 3.48 - 3.39 (m, 1H), 2.77 - 2.58 (m, 1H), 2.43 (d, J = 3.7 Hz, 1H), 2.09 - 1.99 (m, 2H), 1.78 - 1.69 (m, 2H), 1.70 - 1.62 (m, 2H), 1.61 - 1.54 (m, 2H), 1.54 (d, J = 3.6 Hz, 3H), 1.43 (s, 6H), 1.41 (s, 9H), 1.38 (s, 1H), 1.33 - 1.12 (m, 5H), 1.06 (d, J = 3.1 Hz, 3H), 0.94 (s, 3H), 0.92 (d, J = 1.3 Hz, 3H). ESI-MS m/z calc. 593.3829, found 595.58 (M+l)+; Retention time: 2.25 minutes using Method C.

Preparation of Intermediate AJ: 6-(4-(7-(tert-butyl)-5-(4-chloro-3-fluorophenyl)furo[3,2-blpyridine-2- carbonyl)-3,3-dimethylpiperazin-l-yl)-2-methylnicotinic acid

[0279] Intermediate AJ is prepared using General Method 1 with Intermediates A and O, followed by General Procedure 11.

Preparation of Intermediate AK. tert-butyl 4-(5-methoxycarbonyl-6-methyl-2-pyridyl)-2,2- dimethyl- 3-oxo-piperazine-l-carboxylate

Step 1 : 4-(5-methoxycarbonyl-6-methyl-2-pyridyl)-2,2-dimethyl-3-oxo-piperazine-l-carboxylate

[0280] To a solution of tert-butyl 2,2-dimethyl-3-oxo-piperazine-l-carboxylate (1.60 g, 7.01 mmol) in DMF (lO mL) is added 60% NaH/mineral (280 mg, 7.00 mmol). The mixture is stirred at rt for 20 min, then to it are added TBAI (70.0 mg, 0.190 mmol) and tert-butyl 2,2-dimethyl-3-oxo- piperazine-l-carboxylate (1.60 g, 7.01 mmol). The mixture is stirred at rt for lh under nitrogen, the temperature is raised to 60 °C and then stirred for 3h. After cooling to RT, the mixture is neutralized with formic acid, diluted with EtOAc (80 mL), washed with water and brine consecutively, dried over sodium sulfate and filtered. The filtrate is concentrated to dryness under reduced pressure and the residue is purified on Biotage SNAP 50g silica gel cartridge eluting with EtOAc/hexanes 0-30% in 20CV to obtain tert-butyl-4-(5-methoxycarbonyl-6- methyl- 2- pyridyl)-2,2-dimethyl-3-oxo-piperazine-l-carboxylate (755 mg, 37%) as a yellowish solid. ESI- MS m/z calc. 377.19507, found: 378.77 Retention time: 1.9 minutes using Method C.

Step II: methyl 6-(3,3-dimethyl-2-oxo-piperazin-l-yl)-2-methyl-pyridine-3-carboxylate (Dihydrochloride salt)

[0281] To a solution of tert-butyl 4-(5-methoxycarbonyl-6-methyl-2-pyridyl)-2,2-dimethyl-3-oxo- piperazine- 1-carboxylate (755 mg) in DCM (2 mL) is added 4M HCI/dioxane (3.00 mL of 4 M, 12.0 mmol). The mixture is stirred at rt for 4 h. Then the mixture is filtered and the solid is dried in vacuo to provided methyl 6-(3,3-dimethyl-2-oxo-piperazin-l-yl)-2-methyl-pyridine-3- carboxylate (Dihydrochloride salt) (628 mg, 90%) as a white solid. ESI-MS m/z calc. 277.14264, found: 278.68 Retention time: 0.56 minutes using Method C.

Example 1. Preparations of chemical entities.

[0282] Methods of synthesizing representative examples using the disclosed schemes and intermediates are presented in the following examples.

Example 1.1. Preparation of 4-[7-tert-butyl-5-(4-chloro-3-fluorophenyl)furo[3,2-blpyridine-2- carbonyll-3,3-dimethyl-N-(lH-pyrazol-4-ylsulfonyl)piperazine-l-carboxamide (1-12).

[0283] CDI (33.8 mg, 0.208 mmol) is added to a stirred solution of [7-tert-butyl-5-(4-chloro-3- fluoro-phenyl)furo[3,2-b]pyridin-2-yl]-(2,2-dimethylpiperazin-l-yl)methanone (Hydrochloride salt) (100 mg, 0.208 mmol) and DIPEA (56.5 mg, 76.2 μί, 0.437 mmol) in Acetonitrile (1.0 mL). The reaction is stirred at rt for 5 hours.

[0284] To the reaction are added lH-pyrazole-4-sulfonamide (91.91 mg, 0.6246 mmol), followed by DBU (95.1 mg, 93.4 μί, 0.625 mmol). The reaction mixture is then stirred at 60°C for overnight and filtered. The filtrate is diluted in NM P (total of 2 mL) and is directly submitted to the analytical group for reverse-phase HPLC purification using CH₃CN-Water-Ammonium Bicarbonate. The recovered fractions are combined and lyophilized to give 4-[7-tert-butyl-5-(4- chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-N-(lH-pyrazol-4- ylsulfonyl)piperazine-l-carboxamide (24.97 mg, 19%) as a white solid. ESI-MS m/z calc. 616.1671, found 617.25 (M+l)+; Retention time: 3.52 minutes. ^XH NM R (400 MHz, DMSO-d6) δ 13.51 (s, 1H, br), 10.73 (s, 1H, br), 8.27 (s, 1H, br), 8.17 (dd, J = 11.0, 2.0 Hz, 1H), 8.07 - 8.00 (m, 1H), 7.83 (s, 1H), 7.82 - 7.76 (m, 1H, br), 7.72 (t, J = 8.1 Hz, 1H), 7.51 (s, 1H), 3.94 - 3.80 (m, 2H), 3.60 - 3.42 (m, 4H), 1.53 (s, 9H), 1.46 (s, 6H).

Examplel.2. Preparation of 2-[4-[7-tert-butyl-5-(4,4-dimethylcvclohexyl)furo[3,2-blpyridine-2- carbonyll-3,3-dimethyl-piperazin-l-yll-5-methyl-pyridine-4-carboxylic acid (1-6)

Step I:

[0285] To a solution of 7-tert-butyl-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridine-2-carboxylic acid (100 mg, 0.3036 mmol) and ethyl 2-(3,3-dimethylpiperazin-l-yl)-5-methyl-pyridine-4- carboxylate (84.21 mg, 0.3036 mmol) in DMF (800.0 μί) is added DIPEA (196.2 mg, 264.4 μί, 1.518 mmol) and HATU (142.5 mg, 0.3749 mmol) and stirred for lh. The reaction mixture is added water, extracted twice with EtOAc, the combined organic extracts are dried over sodium sulfate, filtered, and the solvent is removed under reduced pressure.

Step II:

[0286] The residue is dissolved in dioxane (1.0 mL), treated with NaOH (455 μί of 2 M, 0.911 mmol) and stirred for two days. The reaction mixture is then brought to pH = 3 using 6 M HCI, the aqueous phase is extracted twice with EtOAc, the combined organic extracts are dried over sodium sulfate, filtered, and the solvent is removed under reduced pressure. The residue is taken up in DMSO and purified using reverse phase preparative HPLC to afford, after lyophilization, 2-[4-[7-tert-butyl-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-piperazin-l-yl]-5-methyl-pyridine-4-carboxylic acid (13 mg, 7%). ^XH NM (400 MHz, DMSO-d6) δ 8.05 (s, 1H), 7.43 (s, 1H), 7.16 (s, 1H), 6.89 (s, 1H), 4.04 (t, J = 5.6 Hz, 2H), 3.88 (s, 2H), 3.67 - 3.53 (m, 2H), 2.80 - 2.64 (m, 1H), 2.29 (s, 3H), 1.83 - 1.65 (m, 4H), 1.51 (s, 6H), 1.47 (s, 11H), 1.35 (dt, J = 13.9, 6.9 Hz, 2H), 0.98 (s, 3H), 0.96 (s, 3H). ESI-MS m/z calc. 560.33624, found 561.3 (M+l)+; Retention time: 3.62 minutes using Method A.

Example 1.3. 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2-carbonyll-3,3- dimethyl-l-(2H-tetrazol-5-yl)piperazin-2-one (1-14)

[0287] Step I: To a solution of 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2- carbonyl]-3,3-dimethyl-piperazin-2-one (Dihydrochloride salt) (Intermediate I) (500 mg, 0.942 mmol) in DM F (10 mL) is added 60% NaH (64.0 mg, 1.60 mmol) and the reaction is stirred at 0^°C for 30 min. Then to it is added carbononitridic bromide (169.5 mg, 163.1 μί, 1.600 mmol) at 0 °C and the mixture is stirred at rt for 6h. The volatiles are removed under reduced pressure and the residue is dissolved in DCM. Some solvent is removed on rotavap (precipitation of the desired product). The product is triturated in DCM and washed with cold DCM to give 4-[7-tert- butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-2-oxo- piperazine-l-carbonitrile (445 mg, 95%) as a light yellow solid. ESI-MS m/z calc. 482.1521, found 485.08 (M+l)+; Retention time: 4.31 minutes using Method A.

[0288] Step II: To a solution of 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2- carbonyl]-3,3-dimethyl-2-oxo-piperazine-l-carbonitrile (440 mg, 0.886 mmol) in DMF (9.8 mL)/ iPrOH (6.5 mL)/ water (3.3 mL) are added NaN₃ (172.7 mg, 2.657 mmol) and ZnBr₂ (199.7 mg, 47.54 μί, 0.8865 mmol). The mixture is stirred at 60 °C for 2h (start at 14h20).

[0289] The reaction mixture is then diluted with EtOAc, washed with water and brine consecutively, dried over sodium sulfate, filtered and concentrated to dryness. The recovered crude compound is purified on Biotage SNAP 50 g silica gel cartridge eluting with 0-15% MeOH/ DCM (loading with DCM). Desired fractions are concentrated and the residue is triturated with EtOAc to obtain upon lyophilization 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2- carbonyl]-3,3-dimethyl-l-(2H-tetrazol-5-yl)piperazin-2-one (148.1 mg, 32%) 1H NM (400 M Hz, DMSO-d6) δ 8.18 (dd, J = 11.0, 2.1 Hz, 1H), 3.34 - 3.29 (m, 10H), 8.11 - 7.99 (m, 1H), 7.85 (s, 1H), 7.76 - 7.69 (m, 1H), 7.66 (s, 1H), 4.00 (s, 2H), 3.93 (s, 2H), 1.82 (s, 6H), 1.53 (s, 9H). ESI-MS m/z calc. 525.1691, found 528.1 (M+l)+; Retention time: 3.96 minutes using Method A.

Example 1.4. 3-[4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2-blpyridine-2-carbonyll-3,3-dimethyl-2- oxo-piperazin-l-yll-2,2-dimethyl-propanoic acid (1-15)

[0290] A solution of 4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-piperazin-2-one (200 mg, 0.4546 mmol) in DM F (2 mL) is treated with sodium hydride (24 μί, 0.900 mmol) and the solution is allowed to stir for 30 min. The mixture is then treated with a solution of methyl 3-chloro-2,2-dimethyl-propanoate (95.8 mg, 0.636 mmol) in DMF (0.3 mL). The mixture is then treated with tetrabutylammoniumiodide (10.4 mg, 0.0282 mmol) and allowed to stir at rt overnight. It is then treated with a further portion of NaH (60 mg) and allowed to stir for 30 min, followed by the addition of the alkyl chloride (120 mg). The mixture is allowed to continue stirring at rt. It is then heated at 60°C overnight. The mixture is quenched with sat NH₄CI and extracted with EtOAc (X3). The combined organic layers are washed (H₂0, brine), dried (Na₂S0₄) and concentrated in vacuo. The residue is dissolved in DMSO and purified by preparative HPLC (CH3CN-H20-formic acid). The appropriate tubes are lyophylized to give a colorless fluffy solid. 3-[4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-2-oxo-piperazin-l-yl]-2,2-dimethyl-propanoic acid (21.1 mg) 1H NMR (400 MHz, DMSO-d6) δ 12.44 (s, 1H), 8.20 - 8.11 (m, 2H), 7.79 (s, 1H), 7.58 (s,lH), 7.58 - 7.51 (m, 2H), 3.81 (d, J = 5.2 Hz, 2H), 3.63 (s, 2H), 3.57 (t, J = 4.8 Hz, 2H), 1.72 (s, 6H), 1.52 (s, 9H), 1.12 (s, 6H). ESI- MS m/z calc. 539.2187, found 540.34 (M+l)+; Retention time: 3.95 minutes using Method A.

Example 1.5. 2-[4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2-blpyridine-2-carbonyll-3,3-dimethyl- piperazin-l-yll-N-methylsulfonyl-pyrimidine-5-carboxamide (1-45)

[0291] EDCI (41.6 mg, 0.217 mmol), DMAP (59.6 mg, 0.488 mmol), 2-[4-[7-tert-butyl-5-(4- chlorophenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-l-yl]pyrimidine-5- carboxylic acid (102 mg, 0.161 mmol) and methanesulfonamide (18.9 mg, 0.199 mmol) are added to DCM (3.2 mL) and the mixture is stirred overnight at room temperature. The reaction mixture is concentrated and purified by reverse phase preparative HPLC. 2-[4-[7-tert-butyl-5- (4-chlorophenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-l-yl]-N-methylsulfonyl- pyrimidine-5-carboxamide (52.4 mg, 50%) ESI-MS m/z calc. 624.19214, found 626.39 (M+l)+; Retention time: 2.0 minutes using Method C. ^XH NMR (400 M Hz, DMSO-d6) δ 11.98 (s, IH), 8.84 (s, 2H), 8.20 - 8.06 (m, 2H), 7.75 (s, IH), 7.56 - 7.52 (m, 2H), 7.52 (s, IH), 4.17 - 3.96 (m, 4H), 3.83 - 3.67 (m, 2H), 3.27 (s, 3H), 1.55 - 1.47 (m, 15H).

Example 1.6. Preparation of 2-[[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2- carbonyll-3,3-dimethyl-piperazine-l-carbonyllaminol-2-methyl-propanoic acid (1-106).

Step I: methyl -2-[[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]- 3,3-dimethyl-piperazine-l-carbonyl]amino]-2-methyl-propanoate [0292] A solution of [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]-(2,2- dimethylpiperazin-l-yl)methanone (Hydrochloride salt) (202.4 mg, 0.4213 mmol) and pyridine (170 μί, 2.10 mmol) in DCM (4 mL) is cooled to 0°C and treated rapidly with phosgene (360 μί of 15 %w/v in toluene, 0.546 mmol) in one portion. The resulting solution is stirred at 0°C for 30 min, and DIPEA (290 μί, 1.665 mmol) is added The reaction is stirred overnight, the solvent evaporated, the residue is dissolved in EtOAc and washed with aqueous 0.5N HCI solution, extracted with EtOAc (3X 20 mL) the combined organic extracts are dried (Na₂S0₄), filtered and concentrated. The crude is used without further purification.

Step II: 2-[[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-piperazine-l-carbonyl]amino]-2-methyl-propanoic acid

[0293] To a solution of crude methyl 2-[[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2- b]pyridine-2-carbonyl]-3,3-dimethyl-piperazine-l-carbonyl]amino]-2-methyl-propanoate (216 mg, 0.369 mmol) in dioxane (3 mL) is added NaOH (200 μΐ of 4 M, 0.800 mmol) and the solution is heated at 40°C overnight. The reaction is acidified with 0.5 M HCI and extracted with EtOAc (X3). The combined organic layers are washed (H₂0, brine), dried (Na₂S0₄) and concentrated in vacuo. The residue is dissolved in DMSO and purified by preparative HPLC (CH₃CN-H₂0-formic acid). The appropriate tubes are lyophylized to give a colorless fluffy solid (105.7 mg, 49%). ^XH NM (400 MHz, DMSO-d6) δ 12.03 (s, 1H), 8.18 (d, J = 11.3 Hz, 1H), 8.04 (d, J = 8.5 Hz, 1H), 7.84 (s, 1H), 7.73 (t, J = 8.1 Hz, 1H), 7.53 (d, J = 1.2 Hz, 1H), 6.30 (s, 1H), 3.91 (t, J = 5.6 Hz, 2H), 3.53 (d, J = 4.5 Hz, 4H), 1.54 (s, 9H), 1.49 (s, 6H), 1.37 (s, 6H). ESI-MS m/z calc. 572.22015, found 572.65 (M+l)+; Retention time: 3.74 minutes using Method A.

Example 1.7. Preparation of 2-[4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2-blpyridine-2-carbonyll- 3,3-dimethyl-piperazine-l-carbonylloxy-2-methyl-propanoic acid (1-110)

Step I: [7-tert-butyl-5-(4-chlorophenyl)furo[3,2-b]pyridin-2-yl]-(2,2-dimethylpiperazin-l- yl)methanone

[0294] A mixture of (200 mg, 0.470 mmol) and TEA (131 μί, 0.939 mmol) are suspended in MeCN (2 mL) and CDI (76.1 mg, 0.470 mmol) is then added at room temperature. The mixture is allowed to stir at RT for 2 h. To the reaction mixture is added methyl 2-hydroxy-2-methyl-propanoate (166.3 mg, 161.0 μί, 1.41 mmol) and DBU (210.5 μί, 1.41 mmol). The reaction is then stirred at 60°C overnight. The mixture is treated again with DBU (210.5 μί, 1.41 mmol) and the whole is stirred over the weekend at 60°C and again overnight at 90°C. A further portion of DBU is added (210.5 μί, 1.41 mmol) and the reaction is stirred at 110°C overnight (no progression). Water and DCM are added and the mixture stirred for 5 minutes. The phases are separated using a phase separator cartridge. The filtrate is concentrated and the crude residue is dissolved in NMP and purified on a Gemini-NX 250-21 column (Isco Combiflash EZ PREP) eluting with 65-95% CH3CN (0.1% Formic Acid)/Water (0.1% Formic Acid) on a 30 minutes run. Desired fractions are combined and lyophilized to give: (2-methoxy-l,l-dimethyl-2-oxo-ethyl) 4-[7-tert-butyl- 5-(4- chlorophenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazine-l-carboxylate (28 mg, 10%) ESI-MS m/z calc. 569.22925, found 570.33 (M+l)+; Retention time: 1.13 minutes using Method J.

Step II: 2-[4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl- piperazine-l-carbonyl]oxy-2-methyl-propanoic acid

[0295] An aqueous solution of LiOH (36.84 μί of 2.0 M, 0.0737 mmol) is added to a stirred heterogenous solution of (2-methoxy-l,l-dimethyl-2-oxo-ethyl) 4-[7-tert-butyl-5-(4- chlorophenyl)furo[3,2-b]pyridine- 2-carbonyl]-3,3-dimethyl-piperazine-l-carboxylate (28.0 mg, 0.0491 mmol) in EtOH (0.5 mL) and THF (0.5 mL). The reaction is stirred at rt for overnight. LiOH (24.6 μί of 2.0 M, 0.0491 mmol) is added and the reaction stirred 2 minutes at 70 and then RT overnight. The reaction mixture is quenched with HCI (20.5 μί of 6.0 M, 0.123 mmol) and the resulting solution is injected on a Gemini-NX 250-21 column (Isco Combiflash EZ PREP) eluting with 60-90% CH3CN (0.1% Formic Acid)/Water (0.1% Formic Acid) over 30 minutes. Desired fractions are combined and lyophilized to give 2-[4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2- b]pyridine-2-carbonyl]- 3,3-dimethyl-piperazine-l-carbonyl]oxy-2-methyl-propanoic acid (12.840 mg, 46%) ^XH NMR (400 MHz, DMSO-d6) δ 12.63 (s, 1H), 8.15 (d, J = 8.1 Hz, 2H), 7.78 (s, 1H), 7.56 (d, J = 8.2 Hz, 2H), 7.52 (s, 1H), 3.92 - 3.87 (m, 1H), 3.63 - 3.55 (m, 2H), 3.52 (s, 1H), 3.47 (s, 1H), 3.31 - 3.24 (m, 1H), 1.56 - 1.46 (m, 21H). ESI-MS m/z calc. 555.2136, found 556.38 (M+l)+; Retention time: 4.01 minutes as a white solid using Method A. Example 1.8. Preparation of 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2- carbonyll-3,3-dimethyl-piperazin-l-yll-2-fluoro-pyridine-3-carboxylic acid and 2-[4-[7-tert-butyl-5-(4- chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2-carbonyll-3,3-dimethyl-piperazin-l-yll-6-fluoro- pyridine-3-carboxylic acid (1-52 and 1-53)

[0296] A solution of [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl] -(2,2- dimethylpiperazin-l-yl)methanone (Hydrochloride salt) (Intermediate D) (612 mg, 1.27 mmol) and 2,6-difluoropyridine-3-carboxylic acid (243 mg, 1.53 mmol) in NMP (6 mL) containing DIPEA (1.00 mL, 5.74 mmol) is heated at 120°C overnight. The reaction mixture is treated with formic acid (1 mL) filtered, and purified by preparative HPLC. 2D NOESY experiments suggested the indicated regiochemistry.

[0297] 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl- piperazin-l-yl]-2-fluoro-pyridine-3-carboxylic acid (26.7 mg, 3%) ^XH NM (400 MHz, DMSO-d6) δ 12.61 (s, 1H), 8.18 (dd, J = 11.0, 2.1 Hz, 1H), 8.15 - 8.07 (m, 1H), 8.04 (dd, J = 8.4, 2.0 Hz, 1H), 7.84 (s, 1H), 7.72 (t, J = 8.1 Hz, 1H), 7.56 (s, 1H), 6.62 (s, 1H), 4.12 (t, J = 5.6 Hz, 2H), 3.93 (s, 2H), 3.68 (t, J = 5.7 Hz, 2H), 1.55 (s, 9H), 1.54 (s, 6H). ESI-MS m/z calc. 582.1845, found 583.44 (M+l)+; Retention time: 4.25 minutes using Method A.

[0298] 2-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl- piperazin-l-yl]-6-fluoro-pyridine-3-carboxylic acid (337.6 mg, 45%) ^XH NM R (400 MHz, DMSO- d6) δ 12.92 (s, 1H), 8.21 - 8.11 (m, 2H), 8.04 (dd, J = 8.5, 2.0 Hz, 1H), 7.83 (s, 1H), 7.71 (t, J = 8.1 Hz, 1H), 7.55 (s, 1H), 6.45 (dd, J = 8.2, 3.1 Hz, 1H), 4.14 - 4.00 (m, 2H), 3.63 (s, 2H), 3.52 (dd, J = 6.6, 4.3 Hz, 2H), 1.55 (s, 6H), 1.53 (s, 9H). ESI-MS m/z calc. 582.1845, found 583.68 (M+l)+; Retention time: 4.41 minutes using Method A.

Example 1.9. Preparation of 3-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2- carbonyll- 3,3-dimethyl-piperazin-l-yll-4-ethoxy-cyclobut-3-ene-l,2-dione (1-90)

[0299] To a solution of [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]-(2,2- dimethylpiperazin- l-yl)methanone (Dihydrochloride salt)(lntermediate D) (300 mg, 0.580 mmol) and NEt₃ (242.7 μί, 1.74 mmol) in ethanol is added 3,4-diethoxycyclobut-3-ene-l,2-dione (170.1 μί, 1.16 mmol) and the solution is heated to reflux for 3h. Solvent is removed under reduced pressure and the crude compound is triturated in diethyl ether to give 3-[4-[7-tert-butyl-5-(4- chloro-3-fluoro-phenyl)furo[3,2- b]pyridine-2- carbonyl]-3,3-dimethyl-piperazin-l-yl]-4-ethoxy- cyclobut-3-ene-l,2-dione as a light brown solid (226 mg, 68%). ESI-MS m/z calc. 567.1936, found 570.37 (M+l)+; Retention time: 4.26 minutes using Method A. This material is of sufficient purity for the preparation of other analogues. A portion of the material (12 mg) is dissolved in NMP and purified on a Gemini-NX 250-21 column (Isco Combiflash EZ PREP) eluting with 60-90% CH3CN (0.1% Formic Acid)/Water (0.1% Formic Acid) over 30 minutes. The desired fractions are combined and lyophilized to give 4.5 mg of pure 3-[4-[7-tert-butyl-5-(4-chloro-3-fluoro- phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-l-yl]-4-ethoxy-cyclobut-3-ene- 1,2-dione. ESI-MS m/z calc. 567.1936, found 570.37 (M+l)⁺; Retention time: 4.32 minutes as a white solid using Method A.

Example 1.10. 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2-carbonyll-l-(2,5- dimethyltriazol-4-yl)-3,3-dimethyl-piperazin-2-one (1-66)

[0300] A solution of 4-bromo-2,5-dimethyl-triazole (107 mg, 0.611 mmol), 4-[7-tert-butyl-5-(4- chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-2-one (200 mg, 0.4368 mmol) (Intermediate I) and K₃P0₄ (185 mg, 0.873 mmol) in DMF (2.2 mL) is charged in a sealed 4 mL vial, flushed with nitrogen, added iodocopper (87.3 mg, 0.459 mmol) and Ν,Ν'- dimethylethane-l,2-diamine (46 mg, 55 μί, 0.524 mmol), and is flushed once more with nitrogen. The reaction is stirred at 100 °C for three hours. The mixture is then filtered and purified using reverse phase prep. HPLC to afford 4-[7-tert-butyl-5-(4-chloro-3-fluoro- phenyl)furo[3,2-b]pyridine-2-carbonyl]-l-(2,5-dimethyltriazol-4-yl)-3,3-dimethyl-piperazin-2-one (110 mg, 45%). 1H NM (400 MHz, DMSO-d6) δ 8.15 (dd, J = 11.0, 2.1 Hz, 1H), 8.07 - 7.97 (m, 1H), 7.82 (s, 1H), 7.72 - 7.66 (m, 1H), 7.63 (s, 1H), 4.02 (s, 5H), 3.92 (d, J = 5.2 Hz, 2H), 2.04 (s, 3H), 1.80 (s, 6H), 1.50 (s, 9H). ESI-MS m/z calc. 552.2052, found 554.64 (M+l)+; Retention time: 4.29 minutes, Method J.

Example 1.11. 2-r4-r4-r7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furor3,2-blpyridine-2-carbonyll-3,3- dimethyl-2-oxo-piperazin-l-yllpyrazol-l-yll-2-methyl-propanoic acid (1-28)

Step I: Ethyl 2-[4-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]- 3,3-dimethyl-2-oxo-piperazin-l-yl]pyrazol-l-yl]-2-methyl-propanoate

[0301] To a solution of 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]- 3,3-dimethyl-l-(lH-pyrazol-4-yl)piperazin-2-one (125 mg, 0.239 mmol) (Intermediate AH) in DMF (2 mL) are added ethyl 2-bromo-2-methyl-propanoate (70.0 mg, 0.359 mmol) and Cs₂C0₃ (116 mg, 0.358 mmol). The mixture is stirred at 55 °C for 2h. It is then diluted with water, stirred at rt for 20 min, filtered. The solid is washed with water, dried in vacuo. The residue is purified by flash column chromatography eluting with EtOAc/hexanes 0-50% to obtain ethyl 2-[4-[4-[7- tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-2-oxo- piperazin-l-yl]pyrazol-l-yl]-2-methyl-propanoate (125 mg, 82%) as a white solid. LC-MS (ESI): 638.51 (M+H)⁺; calc. 638.2545. RT: 2.22 min, Method C.

Step II: 2-[4-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-2-oxo-piperazin-l-yl]pyrazol-l-yl]-2-methyl-propanoic acid

[0302] To a solution of ethyl 2-[4-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2- carbonyl]-3,3-dimethyl-2-oxo-piperazin-l-yl]pyrazol-l-yl]-2-methyl-propanoate (125 mg) in dioxane (3 mL) is added 1M KOH (400 μΐ of 1 M, 0.400 mmol). The mixture is stirred at 80 °C for 3h. It is then neutralized with Resin Amberlite IR120 (H), filtered. The filtrate is concentrated to dryness and the residue is purified using reverse-phase prep-HPLC C18 column eluting with ACN/water/formic acid to provide 2-[4-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2- b]pyridine-2-carbonyl]-3,3-dimethyl-2-oxo-piperazin-l-yl]pyrazol-l-yl]-2-methyl-propanoic acid (40 mg, 32%) as a white solid. ^XH NMR (400 M Hz, DMSO-d6) δ 8.25 - 8.09 (m, 2H), 8.05 - 7.95 (m, 1H), 7.82 (s, 1H), 7.75 - 7.65 (m, 2H), 7.61 (s, 1H), 4.08 - 3.83 (m, 4H), 1.77 (s, 6H), 1.70 (s, 6H), 1.52 (s, 9H). LC-MS (ESI): 610.30 (M+H)⁺; calc. 610.2232. RT: 4.00 min, using Method A.

Example 1.12. 2-[4-[7-f-erf-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2-carbonyll-3,3- dimethyl-piperazin-l-yll-A/-methylsulfonyl-pyrimidine-5-carboxamide (1-26)

[0303] EDCI (15.2 mg, 0.0793 mmol), DMAP (21.1 mg, 0.173 mmol), 2-[4-[7-ferf-butyl-5-(4-chloro-3 -fluoro-phenyl)furo[3,2-b]pyridine- 2-carbonyl]-3,3-dimethyl-piperazin-l-yl]pyrimidine-5- carboxylic acid (34.6 mg, 0.0580 mmol) and methanesulfonamide (6.6 mg, 0.069 mmol) are added to DCM (1.2 mL) and the mixture is stirred for 18 h at room temperature. The mixture is then concentrated under reduced pressure. The residue is taken up in DMSO and purified by reverse phase Prep HPLC (Gemini column, 59-79 % ACN in water (0,1% HCOOH modifier) in 20 minutes) to afford 2-[4-[7-ferf-butyl-5-(4-chloro-3-fluoro- phenyl)furo[3,2-b]pyridine-2- carbonyl]-3,3-dimethyl-piperazin-l-yl]-/V-methylsulfonyl-pyrimidine-5-carboxamide (8.8 mg, 23%). ESI-MS m/z calc. 642.18274, found 643.61 (M+l)+; Retention time: 2.04 minutes using Method C. ^XH NM R (400 MHz, DMSO-d₆) δ 11.98 (s, 1H), 8.83 (s, 2H), 8.15 (dd, J = 11.0, 2.1 Hz, 1H), 8.05 - 7.96 (m, 1H), 7.81 (s, 1H), 7.75 - 7.62 (m, 1H), 7.53 (s, 1H), 4.27 - 3.92 (m, 4H), 3.78 (t, J = 5.7 Hz, 2H), 3.23 (s, 3H), 1.73 - 1.31 (m, 15H).

Example 1.13. 6-(4-(7-(tert-butyl)-5-(4-chloro-3-fluorophenyl)furo[3,2-blpyridine-2-carbonyl)-3,3- dimethylpiperazin-l-yl)-2-methylnicotinonitrile (1-40)

[0304] A mixture of [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]-(2,2- dimethylpiperazin-l-yl)methanone (Intermediate D) (65.0 mg, 0.146 mmol), 6-chloro-2-methyl- pyridine-3-carbonitrile (Intermediate P) (22.4 mg, 0.147 mmol) and cesium carbonate (52.5 mg, 0.161 mmol) in NMP (845 μί) is heated at 100°C for 90 minutes. The mixture is cooled down to room temperature. Then, water is added (50 mL), followed by a saturated NH₄CI aqueous solution (5.0 mL). The mixture is extracted twice with a 1:1 mixture (10 mL) of hexanes and EtOAc. Then, the combined organic layers are dried over MgS0₄, filtered and concentrated under reduced pressure. The obtained residue is purified by flash chromatography on silica gel eluting with mixtures of hexanes and EtOAc (0-100%) to afford the desired 6-[4-[7-tert-butyl-5- (4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-l-yl]-2-methyl- pyridine-3-carbonitrile (53.1 mg, 56%) as a yellow residue. ^XH NM (400 MHz, Chloroform-d) δ 7.84 (dt, J = 10.4, 1.6 Hz, 1H), 7.77 - 7.71 (m, 1H), 7.65 - 7.56 (m, 2H), 7.50 (td, J = 8.0, 1.6 Hz, 1H), 7.43 (d, J = 1.3 Hz, 1H), 6.31 (d, J = 8.8 Hz, 1H), 4.16 - 4.07 (m, 2H), 3.99 (s, 2H), 3.72 (t, J = 5.6 Hz, 2H), 2.59 (s, 3H), 1.65 (s, 6H), 1.57 (s, 9H). ESI-MS m/z calc. 559.215, found 560.25 (M+l)+; Retention time: 4.92 minutes using Method A.

Example 1.14. (7-(tert-butyl)-5-(4-chloro-3-fluorophenyl)furo[3,2-blpyridin-2-yl)(2,2-dimethyl-4-(6- methyl-5-(2H-tetrazol-5-yl)pyridin-2-yl)piperazin-l-yl)methanone (1-39)

[0305] Dibutyltin oxide (10.0 mg, 0.0402 mmol) and trimethylsilyl azide (45.0 μί, 0.342 mmol) are added to a solution of 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2- carbonyl]-3 -dimethyl-piperazin-l-yl]-2-methyl-pyridine-3-carbonitrile (52.0 mg, 0.0793 mmol) in anhydrous 1,4-dioxane (500 μί). The reaction mixture is subjected to microwave irradiation in a tightly sealed glass vessel for 2 hours at 140°C (50% conversion to the title compound is observed by LCMS). The mixture is subjected again to microwave irradiation for 2 hours at 140°C (75% conversion is observed by LCMS) and then at 170°C under microwave irradiation for 1 hour (almost full conversion is observed by LCMS). The mixture is cooled down to room temperature and MeOH (2.0 mL) is added. The mixture is then concentrated to dryness under reduced pressure. The crude residue is taken up in DMSO (2.0 mL), filtered through a 0.2 μιη filter disk and is purified by prep-HPLC affording, after lyophilization, the desired (7-(tert-butyl)-5-(4- chloro-3-fluorophenyl)furo[3,2-b]pyridin-2-yl)(2,2-dimethyl-4-(6-methyl-5-(2H-tetrazol-5- yl)pyridin-2-yl)piperazin-l-yl)methanone (19.0 mg, 35%) as a white solid. ^XH NMR (400 M Hz, DMSO-d6) δ 8.18 (dd, J = 11.0, 2.0 Hz, 1H), 8.05 (dd, J = 8.4, 2.0 Hz, 1H), 7.91 - 7.82 (m, 2H), 7.72 (t, J = 8.2 Hz, 1H), 7.57 (s, 1H), 6.69 (d, J = 8.9 Hz, 1H), 4.12 (t, J = 5.6 Hz, 2H), 4.00 (s, 2H), 3.70 (t, J = 5.6 Hz, 2H), 2.62 (s, 3H), 1.55 (s, 15H). ESI-MS m/z calc. 602.23206, found 601.06 (M-l)-; Retention time: 4.21 minutes using Method A.

Example 1.15. [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridin-2-yll-[4-(2-methoxy-3- Pyridyl)-2,2-dimethyl-piperazin-l-yllmethanone (1-57)

[0306] Pd₂(dba)₃ (38.1 mg, 0.042 mmol) and DavePhos (24.6 mg, 0.062 mmol) are added to a N₂ purged solution of [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]-(2,2- dimethylpiperazin-l-yl)methanone (Hydrochloride salt) (llntermediate D) (100 mg, 0.208 mmol), 3-bromo-2-methoxy-pyridine (39.2 mg, 0.208 mmol) and sodium tert-butoxide (219 μί of 2.0 M, 0.437 mmol) in THF. The reaction is stirred in a sealed tube at 100 °C for 3 hours. At room temperature, water is added along with dichloromethane and the phases are separated using a phase separator cartridge. The organic phase is then filtrated through a pre-packed Celite pad with DCM and concentrated under vacuum. The crude residue is dissolved in NM P and purified on a Gemini-NX 250-21 column (Isco Combiflash EZ PREP) eluting with 65-100% CH₃CN (0.1% Formic Acid)/Water (0.1% Formic Acid) over 30 minutes. Selected fractions are combined and lyophilized to give [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]-[4-(2- methoxy-3-pyridyl)-2,2-dimethyl-piperazin-l-yl]methanone (20.6 mg, 17%) as a yellow solid. ESI-MS m/z calc. 550.2147, found 551.26 (M+l)+; Retention time: 3.37 using Method B. ^XH NM R (400 MHz, DMSO-d6) δ 8.18 (dd, J = 11.0, 2.0 Hz, 1H), 8.04 (dd, J = 8.4, 2.0 Hz, 1H), 7.84 (s, 1H), 7.77 - 7.69 (m, 2H), 7.56 (s, 1H), 7.22 - 7.15 (m, 1H), 6.93 (dd, J = 7.6, 4.9 Hz, 1H), 3.89 (s, 3H), 3.84 - 3.76 (m, 2H), 3.30 - 3.23 (m, 2H), 3.11 (s, 2H), 1.57 (s, 6H), 1.54 (s, 9H).

Example 1.16. [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridin-2-yll-[4-(2-hydroxy-3-

Pyridyl)-2,2-dimethyl-piperazin-l-yllmethanone (1-69)

[0307] To a solution of [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]-[4-(2- methoxy-3-pyridyl)-2,2-dimethyl-piperazin-l-yl]methanone (18.7 mg, 0.032 mmol) in dichloroethane (1 mL) is added BBr₃ (113 μΐ of 1.0 M, 0.113 mmol) dropwise. The reaction mixture is stirred 4h at room temperature and then heated to 40°C overnight. At room temperature, saturated NaHC0₃ along with dichloromethane are added and the mixture is stirred for 5 minutes. Phases are separated using a phase separator cartridge and the filtrate is concentrated under vacuum. The crude residue is dissolved in NMP and purified on a Gemini-NX 250-21 column (Isco Combiflash EZ PREP) eluting with 70-100% CH₃CN (0.1% Formic Acid)/Water (0.1% Formic Acid) over 30 minutes. Selected fractions are combined and lyophilized to give [7- tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]-[4-(2-hydroxy-3-pyridyl)-2,2- dimethyl-piperazin-l-yl]methanone (6.60 mg, 38%) as a white solid. ESI-MS m/z calc. 536.19904, found 537.54 (M+l)+; Retention time: 4.14 minutes using Method A. ^XH NMR (400 MHz, DMSO- d6) δ 11.42 (s, 1H, broad), 8.18 (dd, J = 11.0, 2.0 Hz, 1H), 8.08 - 8.00 (m, 1H), 7.84 (s, 1H), 7.76 - 7.67 (m, 1H), 7.55 (s, 1H), 6.96 - 6.91 (m, 1H), 6.68 - 6.61 (m, 1H), 6.14 (t, J = 6.8 Hz, 1H), 3.85 - 3.78 (m, 2H), 3.32 - 3.26 (m, 4H), 1.54 (s, 15H).

Example 1.17. Preparation of 2-[4-[7-tert-butyl-5-(4,4-dimethylcvclohexyl)furo[3,2-blpyridine-2- carbonyll- 3,3-dimethyl-piperazine-l-carbonylloxy-2-methyl-propanoic acid (1-116)

Step I: (2-methoxy-l,l-dimethyl-2-oxo-ethyl) 4-[7-tert-butyl-5-(4,4- dimethylcyclohexyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazine-l-carboxylate

[0308] A solution of [7-tert-butyl-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridin-2-yl]-(2,2- dimethylpiperazin- l-yl)methanone (Hydrochloride salt) (Intermediate E) (300 mg, 0.649 mmol) in DCM (4 mL) is treated with pyridine (183.8 μί, 2.27 mmol) and after 2 min carbonyl dichloride (599 μί of 15 %w/v in toluene, 0.909 mmol) is added rapidly (note exotermic reaction) in one portion. The reaction mixture is stirred at T for 10 min and then treated with methyl 2-hydroxy- 2-methyl-propanoate (104 μί, 0.909 mmol). The resulting solution is allowed to stir for lh. The solvent is evaporated in vacuo, the residue is diluted with EtOAc and acidified with HCI (0.5 M), extracted with EtOAc (3 X 20mL) washed with brine, dried over Na₂S0₄ filtered and concentrated.

Step II: 2-[4-[7-tert-butyl-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridine-2-carbonyl]- 3,3-dimethyl- piperazine-l-carbonyl]oxy-2-methyl-propanoic acid

[0309] The crude material obtained above is dissolved in dioxane (4 mL) and treated with NaOH (162 μΐ of 4 M, 0.648 mmol). The mixture is heated at 40°C overnight. UPLCMS showed incomplete conversion. The temperature is increased to 50°C and a further 200 uL of NaOH are added. After stirring lh, a further 200 uL of NaOH (4M) is added and the temperature increased to 60°C. After a further 7h (incomplete conversion) the reaction is cooled to RT and acidified with 0.5 M HCI and diluted with H₂0. The mixture is extracted with EtOAc (3X20 mL) and washed with brine, dried over Na₂S0₄ filtered and concentrated. The residue is purified by preparative HPLC (MeCN-H20-formic acid) and the appropriate fractions are lyophyllized to give a colorless fluffy solid. (91.6 mg, 25%) ^XH NMR (400 MHz, DMSO-d6) δ 12.62 (s, 1H), 7.40 (s, 1H), 7.16 (s, 1H), 3.85 (t, J = 5.7 Hz, 2H), 3.57 (d, J = 10.6 Hz, 2H), 3.50 (s, 1H), 3.45 (s, 1H), 2.71 (dd, J = 14.1, 10.4 Hz, 1H), 1.85 - 1.61 (m, 4H), 1.48 (s, 12H), 1.45 (s, 9H), 1.33 (td, J = 13.5, 4.2 Hz, 3H), 1.00 (s, 1H), 0.98 (s, 3H), 0.95 (s, 3H). ESI-MS m/z calc. 555.3308, found 556.48 (M+l)+; Retention time: 4.16 minutes using Method A. Example 1.18. Preparation of 3-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2- carbonyll-3,3- dimethyl-piperazin-l-yll-4-(isopropylamino)cyclobut-3-ene-l,2-dione (1-113)

[0310] To a stirred solution of TEA (24.5 μΐ, 0.176 mmol) in EtOH (1 mL) is added 3-[4-[7-tert-butyl- 5-(4- chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-l-yl]-4- ethoxy-cyclobut-3-ene-l,2-dione (100 mg, 0.176 mmol) followed by iPrNH₂ (90.7 μΐ, 1.06 mmol). The reaction is stirred for 3 hours at 50 °C and then at 80°C overnight. A further 6.0 eq of NEt₃ (90.7 μί, 1.06 mmol) (with TEA 24.5 μί, 0.176 mmol)) and the reactions are stirred overnight at 50 °C. The reaction mixture is diluted with NM P and purified on a Gemini-NX 250-21 column (Isco Combiflash EZ PREP) using 0.1% Formic Acid)/Water (0.1% Formic Acid as eluent over 30 minutes. The desired fractions are combined and lyophilized to give a solid (59.26 mg, 57%). ESI-MS m/z calc. 580.2253, found 581.81 (M+l)+; Retention time: 3.94 minutes using Method A. XH NMR (400 MHz, DMSO-d6) δ 8.22 - 8.13 (m, 1H), 8.08 - 8.00 (m, 1H), 7.84 (s, 1H), 7.77 - 7.67 (m, 1H), 7.57 (s, 1H), 7.52 (d, J = 8.4 Hz, 1H), 4.48 - 4.37 (m, 1H), 4.06 - 3.91 (m, 4H), 3.85 (s, 1H), 3.32 (s, 2H), 1.54 (s, 9H), 1.24 - 1.22 (m, 6H), 1.22 - 1.18 (m, 6H).

Example 1.19. Preparation of 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2- carbonyll-3,3-dimethyl- piperazin-l-yll-2,4-dimethyl-pyridine-3-carboxylic acid (1-117)

Step I: ethyl 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-piperazin-l-yl]-2,4-dimethyl-pyridine-3-carboxylate

[0311] To a solution of [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]-(2,2- dimethylpiperazin- l-yl)methanone (Dihydrochloride salt) (Intermediate D) (350 mg, 0.677 mmol) in NM P (5 mL) are added ethyl 6-chloro-2,4-dimethyl-pyridine-3-carboxylate (187 mg, 0.875 mmol) and cesium carbonate (792 mg, 2.43 mmol), and the mixture is heated at 125°C for two days. The reaction mixture is combined with that of a previous batch (200 mg scale). The mixture is diluted with water and extracted three times with EtOAc and the combined organic extracts are washed with water, brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue is purified by flash chromatography to afford the title compound as a yellow foam (197.7 mg). ethyl 6-[4-[7-tert-butyl-5-(4-chloro-3- fluoro- phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-l-yl]-2,4-dimethyl-pyridine-3- carboxylate ^XH NM (400 M Hz, DMSO-d6) δ 8.19 (d, J = 11.0 Hz, 1H), 8.05 (d, J = 8.5 Hz, 1H), 7.85 (s, 1H), 7.73 (t, J = 8.1 Hz, 1H), 7.56 (s, 1H), 6.39 (s, 1H), 4.28 (q, J = 7.1 Hz, 2H), 4.09 (t, J = 5.6 Hz, 2H), 3.94 (s, 2H), 3.65 (t, J = 5.5 Hz, 2H), 2.37 (s, 3H), 2.26 (s, 3H), 1.56 (s, 9H), 1.53 (s, 6H), 1.30 (t, J = 7.1 Hz, 3H). ESI-MS m/z calc. 620.2566, found 621.54 (M+l)+; Retention time: 1.35 minutes using Method J.

Step II: 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl- piperazin-l-yl]-2,4-dimethyl-pyridine-3-carboxylic acid

[0312] A solution of ethyl 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2- carbonyl]-3,3- dimethyl-piperazin-l-yl]-2,4-dimethyl-pyridine-3-carboxylate (400 mg, 0.644 mmol) in tert-butanol (6 mL) is treated with KOtBu (290 mg, 2.58 mmol) and the mixture is heated to 80°C. After stirring for 5 h the reaction is cooled to RT and acidified with HCI (5.0 mL of 0.5 M, 2.50 mmol) and diluted with H₂0. The mixture is extracted with EtOAc x3 and washed with brine, dried over Na₂S0₄ filtered and concentrated. The residue is purified by preparative HPLC (MeCN-H20-formic acid) and the appropriate fractions are lyophyllized to give a colorless fluffy solid. 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2- b]pyridine-2-carbonyl]-3,3- dimethyl-piperazin-l-yl]-2,4-dimethyl-pyridine-3-carboxylic acid (133 mg, 34%) ESI-MS m/z calc. 592.2253, found 593.22 (M+l)+; Retention time: 3.88 minutes using Method A. ^XH NMR (400 MHz, DMSO-d6) δ 12.80 (s, 1H), 8.21 (dd, J = 11.0, 2.1 Hz, 1H), 8.08 (dd, J = 8.5, 2.1 Hz, 1H), 7.87 (s, 1H), 7.75 (t, J = 8.1 Hz, 1H), 7.59 (s, 1H), 6.39 (s, 1H), 4.11 (t, J = 5.7 Hz, 2H), 3.95 (s, 2H), 3.66 (t, J = 5.5 Hz, 2H), 2.41 (s, 3H), 2.30 (s, 3H), 1.58 (s, 9H), 1.55 (s, 6H).

Example 1.20. Preparation of 3-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2- carbonyll-3,3-dimethyl-l,4-diazepan-l-yll-4-hvdroxy-cyclobut-3-ene-l,2-dione (1-108)

Step I: [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]-(2,2-dimethyl-l,4- diazepan-l-yl)methanone

[0313] To a stirred solution of tert-butyl 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2- bjpyridine- 2-carbonyl]-3,3- dimethyl-l,4-diazepane-l-carboxylate (182 mg, 0.326 mmol) in DCM (1 mL) is added TFA (251 μί, 3.26 mmol) and the reaction stirred at RT for 4 hours. A further portion of TFA is added (126 μί, 1.63 mmol) and the reaction stirred for 2 hours. The reaction mixture is concentrated under reduced pressure. Saturated NaHC0₃ (aq) is added along with DCM and the mixture stirred for 5 minutes. The phases are separated using a phase separator and the filtrate is concentrated to give [7-tert-butyl-5-(4- chloro-3 fluoro-phenyl)furo[3,2- b]pyridin-2-yl]-(2,2-dimethyl-l,4-diazepan-l-yl)methanone (143 mg, 96%) ESI-MS m/z calc. 457.19324, found 458.39 (M+l)+; Retention time: 0.68 minutes using Method J.

Step II: 3-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl- l,4-diazepan-l-yl]-4-ethoxy-cyclobut-3-ene-l,2-dione

[0314] To a solution of [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]-(2,2- dimethyl- 1,4-diazepan-l- yl)methanone (120 mg, 0.262 mmol) and Et₃N (106.0 mg, 146.0 μί, 1.048 mmol) in EtOH (400 μί) is added 3,4-diethoxycyclobut-3-ene-l,2-dione (89.2 mg, 0.524 mmol) and the solution is heated to 80°C over the weekend. Et₂0 (5 mL) is added and the compound filtered using a Buchner filter. The solid is washed with diethyl ether to give after drying 3-[4-[7-tert-butyl-5-(4-chloro-3- fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-l,4-diazepan-l-yl]-4-ethoxy-cyclobut-3-ene-l,2-dione (150 mg, 98%) as a beige solid. ESI-MS m/z calc. 581.2093, found 582.48 (M+l)+; Retention time: 1.12 minutes using Method J. Step III: 3-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-l,4-diazepan-l-yl]-4-hydroxy-cyclobut-3-ene-l,2-dione

[0315] To a solution of 3-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2- carbonyl]-3,3- dimethyl-l,4-diazepan-l-yl]-4-ethoxy-cyclobut-3-ene-l,2-dione (150 mg, 0.258 mmol) in EtOH (500 μί) is added an aqueous solution of HCI (256 μί of 6.0 M, 1.55 mmol) and the solution is heated to 50°C for 2 hours. Another portion of HCI (300 μΐ of 3.0 M, 0.900 mmol) is added and the reaction is stirred overnight at 50°C. HCI (515 μί of 1.0 M, 0.515 mmol) is added and the reaction is stirred 2 hours at 50°C and overnight at 60°C. The reaction is dissolved in NM P and purified on a Gemini-NX 250-21 column (Isco Combiflash EZ PREP) eluting with 50- 80% CH3CN (0.1% Formic Acid)/Water (0.1% Formic Acid) over 30 minutes. The desired fractions are combined and lyophilized to give 3-[4-[7-tert-butyl-5-( 4-chloro-3-fluoro- phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-l,4-diazepan-l-yl]-4-hydroxy-cyclobut-3- ene-l,2-dione (13.420 mg, 9%) as a beige solid . ESI-MS m/z calc. 553.178, found 554.33 (M+l)+; Retention time: 3.2 minutes using Method A. ¹H NM R (400 MHz, Chloroform-d) δ 7.92 - 7.78 (m, 1H), 7.79 - 7.66 (m, 1H), 7.57 (s, 1H), 7.53 - 7.47 (m, 1H), 7.44 (s, 1H), 4.05 - 4.00 (m, 2H), 3.93 (s, 2H), 3.88 - 3.83 (m, 2H), 2.25 - 2.20 (m, 2H), 1.68 (s, 6H), 1.56 (s, 9H).

Example 1.21. Preparation of 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2- carbonyll-3,3-dimethyl-piperazin-l-yll-2-chloro-pyridine-3-carboxylic acid and 2-[4-[7-tert-butyl-5-(4- chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2-carbonyll-3,3-dimethyl-piperazin-l-yll-6-chloro- pyridine-3-carboxylic acid (1-29 and 1-30)

Step I: Methyl 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]- 3,3-dimethyl-piperazin-l-yl]-2-chloro-pyridine-3-carboxylate and methyl 2-[4-[7-tert-butyl-5- (4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]- 3,3-dimethyl-piperazin-l-yl]-6- chloro-pyridine-3-carboxylate

[0316] To a solution of [7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2-yl]-(2,2- dimethylpiperazin- l-yl)methanone (Dihydrochloride salt) (Intermediate D) (400 mg, 0.774 mmol) in NMP (3 mL) are added methyl 2,6-dichloropyridine-3-carboxylate (151.4 mg, 0.735 mmol) and DIPEA (320 μί, 1.84 mmol). The mixture is heated at 50 C for 12 h. Then it is diluted with water, stirred at rt for 20 min and filtered. The solid is washed with water and dried in vacuo. The residue is purified by flash chromatography (25g silica gel) eluting with EtOAc/hexanes 0-50% to obtain the major isomer methyl 6-[4-[7-tert-butyl-5- (4-chloro-3-fluoro- phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin- l-yl]-2-chloro- pyridine-3- carboxylate (210 mg, 47%) as a white solid. LC-MS (ESI): 613.73 (M+H)⁺; calc. 613.1784. RT: 2.36 min using Method C. ^XH NMR (400 M Hz, Chloroform-d) δ 8.09 (d, J = 8.7 Hz, 1H), 7.83 (dd, J = 10.3, 2.1 Hz, 1H), 7.74 (ddd, J = 8.4, 2.2, 0.8 Hz, 1H), 7.58 (s, 1H), 7.50 (dd, J = 8.4, 7.4 Hz, 1H), 7.45 (s, 1H), 6.36 (d, J = 8.8 Hz, 1H), 4.15 - 4.05 (m, 2H), 3.94 (s, 2H), 3.87 (s, 3H), 3.73 (t, J = 5.7 Hz, 2H), 1.65 (s, 6H), 1.56 (s, 9H) along with the minor isomer: methyl 2-[4-[7-tert-butyl-5-(4- chloro-3-fluoro- phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-l-yl]-6-chloro- pyridine-3-carboxylate (98 mg). LC-MS (ESI): 613.73 (M+H)⁺; calc. 613.1784. RT: 2.46 min using Method C. ^XH NMR (400 M Hz, Chloroform-d) δ 7.94 (d, J = 8.0 Hz, 1H), 7.82 (dd, J = 10.3, 2.0 Hz, 1H), 7.77 - 7.67 (m, 1H), 7.56 (s, 1H), 7.49 (dd, J = 8.4, 7.5 Hz, 1H), 7.43 (d, J = 1.2 Hz, 1H), 6.72 (d, J = 8.0 Hz, 1H), 4.09 - 3.98 (m, 2H), 3.87 (s, 3H), 3.66 - 3.55 (m, 4H), 1.65 (s, 6H), 1.54 (s, 9H).

Step II:

6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl- piperazin-l-yl]-2-chloro-pyridine-3-carboxylic acid

[0317] To a solution of methyl 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2- carbonyl]-3,3-dimethyl-piperazin-l-yl]-2-chloro-pyridine-3-carboxylate (160 mg, 0.261 mmol) in dioxane (2 mL) is added 1M KOH (500 μΐ of 1 M, 0.500 mmol). The mixture is stirred at 60°C for 4h. Then it is neutralized with Amberlite 120 (H) resin and filtered. The filtrated is concentrated to dryness under reduced pressure. The residue is triturated in hot methanol, and then the resulting solid is dissolved in MeCN-H₂0 and freeze-dried to provide the title compound (150 mg, 93%). ESI-MS m/z calc. 598.15497, found 599.2 (M+l)+; Retention time: 4.35 minutes using Method A. ^XH NM (400 M Hz, DMSO-d6) δ 12.73 (s, 1H), 8.14 (s, 1H), 8.01 (t, J = 9.0 Hz, 2H), 7.80 (s, 1H), 7.67 (t, J = 8.1 Hz, 1H), 7.52 (s, 1H), 6.67 (s, 1H), 4.08 (t, J = 5.6 Hz, 2H), 3.90 (s, 2H), 3.64 (t, J = 5.5 Hz, 2H), 1.51 (s, 15H).

2-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl- piperazin-l-yl]-6-chloro-pyridine-3-carboxylic acid

[0318] To a solution of methyl 2-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2- carbonyl]-3,3-dimethyl-piperazin-l-yl]-6-chloro-pyridine-3-carboxylate (95 mg, 0.155 mmol) in dioxane (2 mL) is added 1M KOH (300 μΐ of 1 M, 0.300 mmol). The mixture is stirred at 75°C for 12h. It is then neutralized with Amberlite IR120 (H) resin and filtered. The filtrate is concentrated to dryness and the residue is purified using reverse-phase prep-HPLC C18 column eluting with ACN/water/formic acid to provide 2-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2- b]pyridine-2-carbonyl]-3,3- dimethyl-piperazin-l-yl]-6-chloro-pyridine-3-carboxylic acid (60 mg, 63%) as a white solid. ESI-MS m/z calc. 598.15497, found 599.2 (M+l)+; Retention time: 4.63 minutes using Method A. ^XH NMR (400 MHz, DMSO-d6) δ 13.08 (s, 1H), 8.13 (dd, J = 11.0, 2.1 Hz, 1H), 8.00 (dd, J = 8.5, 2.0 Hz, 1H), 7.93 (s, 1H), 7.79 (s, 1H), 7.68 (t, J = 8.1 Hz, 1H), 7.51 (s, 1H), 6.78 (d, J = 7.9 Hz, 1H), 4.02 (t, J = 5.4 Hz, 2H), 3.62 (s, 2H), 3.48 (t, J = 5.5 Hz, 2H), 1.52 (s, 6H), 1.50 (s, 9H).

Example 1.22. 2-[4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2-blpyridine-2-carbonyll-3,3-dimethyl- piperazine-l-carbonyllcyclohexanecarboxylic acid (1-41)

[0319] A solution of [7-tert-butyl-5-(4-chlorophenyl)furo[3,2-b]pyridin-2-yl]-(2,2-dimethylpiperazin- l-yl)methanone (Dihydrochloride salt) (Intermediate F) (205 mg, 0.411 mmol), racemic-cis- cyclohexane-l,2-dicarboxylic acid (71 mg, 0.412 mmol) and HATU (174 mg, 0.4576 mmol) in DMF (3 mL) at room temperature is treated with DIEA (290 μί, 1.67 mmol) dropwise. The mixture is stirred for 90 min, then water (1.0 mL) is added slowly. After being stirred for 10 minutes, water (60 mL), EtOAc (5 mL) and hexanes (5 mL) are added. The phases are separated and the aqueous layer is extracted twice with a 1:1 mixture of hexanes and EtOAc (2 X 10 mL). The combined organic layers are dried over MgS0₄, filtered and concentrated under reduced pressure. The crude product is purified by flash chromatography on silica gel eluting with mixtures of DCM and MeOH. Fractions containing the title compound are pooled and concentrated to dryness under reduced pressure. The obtained residue (234 mg) is dissolved in DMSO (1.5 mL) and purified by prep-HPLC eluting with mixtures of acetonitrile and water (0.1% formic acid) affording both stereoisomers of cis- 2-[4-[7-tert-butyl-5-(4-chlorophenyl)furo[3,2- b]pyridine-2-carbonyl]-3,3-dimethyl-piperazine-l-carbonyl]cyclohexanecarboxylic acid (94.5 mg, 36%) as a white solid. ESI-MS m/z calc. 579.25, found 578.35 (M-l)+; Retention time: 4.08 minutes using Method A. ^XH NMR (400 MHz, DMSO-d6) δ 11.38 (brs, 1H), 8.15 (d, J = 8.5 Hz, 2H), 7.77 (s, 1H), 7.59 - 7.46 (m, 3H), 4.01 - 3.89 (m, 2H), 3.86 - 3.47 (m, 4H), 2.50 - 2.36 (m, 1H), 2.31 - 2.09 (m, 1H), 1.90 - 1.66 (m, 3H), 1.64 - 1.17 (m, 20H).

Example 1.23. l-[(4S)-l-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2-carbonyll- 2,2-dimethyl-4-piperidyll-3-methylsulfonyl-urea and l-[(4R)-l-[7-tert-butyl-5-(4-chloro-3-fluoro- phenyl)furo[3,2-blpyridine-2-carbonyll-2,2-dimethyl-4-piperidyll-3-methylsulfonyl-urea (1-101 and I- 103)

[0320] At room temperature, CDI (35.2 mg, 0.217 mmol) is added to a stirred solution of (4-amino- 2,2-dimethyl- l-piperidyl)-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridin-2- yljmethanone (106 mg, 0.217 mmol) and DIPEA (83.0 μί, 0.477 mmol) in acetonitrile (1.0 mL). The reaction is stirred for 2h at RT and then, methanesulphonamide (61.9 mg, 0.651 mmol) followed by DBU (99.1 mg, 97.3 μί, 0.651 mmol) are added. The reaction is stirred at 60°C overnight. After cooling to RT, the mixture is filtered and injected on a Gemini-NX 250-21 column (Isco Combiflash EZ PREP) for purification, eluting with 50-90% CH3CN (0.1% Formic Acid)/Water (0.1% Formic Acid) over 30 min. Selected fractions are combined and lyophilized to give 88 mg of the racemic mixture of enantiomers. This material (22.3 mg) is dissolved in 4mL of methanol and the solution is purified by SCF using a ChiralPak IC column (250X10 mm, 5 μιη) and isocratic method with 25% (50%CH₃CN-50% Isopropanol) and 75% C0₂ with a flow rate of 15 mL/min. The appropriate fractions are concentrated and then dissolved in CH₃CN-H₂0 and lyophilized to give the enantiomers (structures are randomly assigned): [0321] The first eluting compound is l-[(4S)-l-[7-tert-butyl-5-(4-chloro-3-fluoro- phenyl)furo[3,2- b]pyridine-2-carbonyl]-2,2-dimethyl-4-piperidyl]-3-methylsulfonyl-urea (7.56 mg) as a white solid. ^XH NMR (400 MHz, DMSO-d6) δ 9.99 (s, 1H, broad), 8.21 - 8.13 (m, 1H), 8.07 - 8.00 (m, 1H), 7.83 (s, 1H), 7.76 - 7.67 (m, 1H), 7.52 (s, 1H), 6.57 - 6.50 (m, 1H), 3.86 - 3.78 (m, 2H), 3.38 - 3.27 (m, 1H), 3.19 (s, 3H), 2.02 (s, 1H), 1.83 - 1.73 (m, 1H), 1.55 (d, J = 14.2 Hz, 14H), 1.49 - 1.44 (m, 3H). ESI-MS m/z calc. 578.1766, found 579.71 (M+l)+; Retention time: 3.95 minutes using Method A.

[0322] The second eluting compound is l-[(4R)-l-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2- b]pyridine-2-carbonyl]-2,2-dimethyl-4-piperidyl]-3-methylsulfonyl-urea (8.17 mg, 70%). ^XH NMR (400 M Hz, DMSO-d6) δ 9.99 (s, 1H, broad), 8.21 - 8.13 (m, 1H), 8.07 - 8.00 (m, 1H), 7.83 (s, 1H), 7.76 - 7.67 (m, 1H), 7.52 (s, 1H), 6.57 - 6.50 (m, 1H), 3.86 - 3.78 (m, 2H), 3.38 - 3.27 (m, 1H), 3.19 (s, 3H), 2.02 (s, 1H), 1.83 - 1.73 (m, 1H), 1.55 (d, J = 14.2 Hz, 14H), 1.49 - 1.44 (m, 3H). ESI-MS m/z calc. 578.1766, found 580.6 (M+l)+; Retention time: 4.02 minutes as an off-white solid using Method A.

Example 1.24. 3-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2-carbonyll- 3,3- dimethyl-piperazine-l-carbonyllbicvclo[3.1.01hexane-6-carboxylic acid (1-55)

Step I: tert-butyl cyclopent-3-ene-l-carboxylate

[0323] A mixture of 2-methylprop-l-ene (13 mL, 134 mmol) at -78°C is treated with cyclopent-3- ene-1- carboxylic acid (15.0 g, 134 mmol) in Ether (40 mL), followed by sulfuric acid (1.30 mL, 24.4 mmol). The vessel is sealed and the cooling bath removed, and kept behind a blast shield. The mixture is allowed to stir at rt for 4 days. The reaction mixture is cooled to -78°C and poured cautiously into 40 m L Et₂0 and 150 m L sat NaHC0₃. The layers are separated and the aq phase is extracted with Et₂0. The combined organic layers are dried over MgS0₄ and filtered. The solution is concentrated in vacuo and the residue distilled (bp 71-73°C, 20 mbar) to give a colorless liquid in three fractions, all of which are identical by NM . They are combined affording 16.98 g. ^XH NM R (400 M Hz, Chloroform-d) δ 5.64 (s, 2H), 3.12 - 2.94 (m, 1H), 2.61 (s, 2H), 2.59 (s, 2H), 1.45 (s, 10H).

Step II: 3-tert-butyl-6-ethyl bicyclo[3.1.0]hexane-3,6-dicarboxylate and ethyl -3-[4-[7-tert- butyl-5-(4- chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazine-l- carbonyl]bicyclo[3.1.0]hexane- 6-carboxylate

[0324] Following a literature method (2004, Eur. J. Org. Chem. 289), a mixture of tert-butyl cyclopent-3-ene-l-carboxylate (2.00 g, 11.9 mmol) and Rh₂OAc₄ (5.80 mg, 0.0131 mmol) in DCM (10 m L) is treated dropwise over 1.5 h with a solution of ethyl 2-diazoacetate (1.25 m L, 11.9 mmol) in DCM (10 m L). The reaction mixture is concentrated and purified by flash chromatrography to give two fractions. A portion of 100 mg of the mixed fractions is treated with DCM (0.5 m L) and TFA (0.5 m L) and the mixture is allowed to stir for 4h and the solvents removed in vacuo. The residue is treated with [7-tert-butyl-5-(4-chloro-3-fluoro- phenyl)furo[3,2-b]pyridin-2-yl]-(2,2-dimethylpiperazin-l-yl)methanone (Intermediate D) (150 mg, 0.338 mmol), Hunigs base (414 μί, 2.38 mmol) and HATU (180.8 mg, 0.476 mmol) in DM F (3 m L). The reaction is treated with formic acid and purified by preparative HPLC. Obtained 86.2 mg of the desired product. Spectral data (NOESY), suggest that the indicated stereochemistry is correct, (NOE between the protons alpha to both carbonyls). A minor isomer is obtained but not pursued.

Step III : 3-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl- piperazine-l-carbonyl]bicyclo[3.1.0]hexane-6-carboxylic acid

[0325] A solution of ethyl 3-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2- carbonyl]- 3,3-dimethyl-piperazine-l-carbonyl]bicyclo[3.1.0]hexane-6-carboxylate (82.3 mg, 0.132 mmol) in THF (2 m L) is treated with NaOH (60 μΐ of 10 %w/w) and the solution is allowed to stir at rt. The reaction is heated at 40°C overnight and a further portion of NaOH (60 μί of 10 %w/w) is added and stirring continued for the weekend. A further 1 equiv of base is added and stirring continued. The reaction required a total of 4 days. The reaction mixture is cooled to rt, acidified with HCI (0.5 M) and extracted with EtOAc X2. The combined organic layers are washed (H₂0, brine), dried (Na₂S0₄) and concentrated. The residue is dissolved in DMSO and treated with 0.1 mL formic acid and 0.25 mL H₂0, filtered through a 0.45 micron disc and purified by prep HPLC (MeCN-H₂0-formic acid) to give a colorless fluffy solid. 3-[4-[7-tert-butyl-5-(4-chloro- 3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]- 3,3-dimethyl-piperazine-l- carbonyl]bicyclo[3.1.0]hexane-6-carboxylic acid (34.8 mg, 44%). The NM R showed the presence of rotamers as indicated. ^XH NMR (400 M Hz, DMSO-d6) δ 11.95 (s, 1H), 8.17 (dd, J = 11.0, 2.4 Hz, 1H), 8.03 (dd, J = 8.6, 2.3 Hz, 1H), 7.84, 7.83 (s, 1H), 7.72 (td, J = 8.2, 2.4 Hz, 1H), 7.56, 7.51 (s, 1H), 3.92 (dt, J = 11.8, 5.6 Hz, 2H), 3.76 (t, J = 5.6 Hz, 1H), 3.71 (s, 1H), 3.64 (s, 1H), 3.51 - 3.44 (m, 1H), 2.91 - 2.65 (m, 1H), 2.11 - 1.90 (m, 4H), 1.78 (d, J = 3.6 Hz, 2H), 1.64 (d, J = 3.0 Hz, 1H), 1.54 (s, 3H), 1.53, 1.52 (s, 9H), 1.46 (s, 3H). ESI-MS m/z calc. 595.2249, found 596.61 (M+l)+; Retention time: 3.81 minutes using Method A.

Example 1.25. Preparation of (2S,3S,4S,5R,6S)-6-(((ls,4R)-4-(4-(7-(tert-butyl)-5-(4,4- dimethylcvclohexyl)furo[3,2-blpyridine-2-carbonyl)-3,3-dimethylpiperazine-l-carbonyl)-l- methylcvclohexane-l-carbonyl)oxy)-3,4.5-trihvdroxytetrahvdro-2H-pyran-2-carboxylic acid and (2S,3S,4S,5R,6R)-6-(((ls,4S)-4-(4-(7-(tert-butyl)-5-(4,4- dimethylcvclohexyl)furor3,2-blpyridine- 2- carbonyl)-3,3-dimethylpiperazine-l-carbonyl)-l-methylcvclohexane-l-carbonyl)oxy)-3,4.5- trihydroxytetrahvdro-2H-pyran-2-carboxylic acid (1-59 and 1-60)

[0326] A mixture of 4-(4-(7-(tert-butyl)-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridine-2-carbonyl)- 3,3-dimethylpiperazine-l-carbonyl)-l-methylcyclohexane-l-carboxylic acid (Intermediate Al) (152 mg, 0.256 mmol), allyl (3S,4S,6R)-3,4,5,6-tetrahydroxytetrahydropyran-2-carboxylate (120 mg, 0.512 mmol) and PPh₃ (134 mg, 0.511 mmol) are dissolved in THF (2 mL). The mixture is cooled in ice bath and DIAD (100 μί, 0.516 mmol) is added. The mixture is stirred in an ice bath for 1 hour, warmed to room temperature and stirred overnight. Pd(PPh₃)₄ (29.0 mg, 0.0251 mmol) and pyrrolidine (65 μί, 0.78 mmol) are added and the mixture is stirred at room temperature for an additional 2 hours. The crude mixture is evaporated to dryness and the residue is purified by reverse phase HPLC (Gemini column, 45% to 75% ACN in water (0,1% HCOOH modifier) in 40 minutes. The first eluting peak is lyophilised to afford (2S,3S,4S,5R,6S)-6- (((ls,4R)-4-(4- (7-(tert-butyl)-5-(4,4-dimethylcyclohexyl)furo[3,2-b]pyridine-2-carbonyl)-3,3- dimethylpiperazine-l-carbonyl)-l-methylcyclohexane-l-carbonyl)oxy)-3,4,5- trihydroxytetrahydro-2H-pyran-2-carboxylic acid (26.5 mg, 12%) ^XH NM R (400 M Hz, Methanol- d4) δ 7.35 - 7.25 (m, 1H), 7.26 - 7.15 (m, 1H), 5.52 (d, J = 7.8 Hz, 1H), 4.05 - 3.95 (m, 2H), 3.83 (t, J = 5.6 Hz, 2H), 3.76 (s, 1H), 3.72 (s, 1H), 3.63 - 3.56 (m, 1H), 3.52 (broad s, 1H), 3.47 - 3.40 (m, 1H), 3.36 (t, J = 8.4 Hz, 1H), 2.78 - 2.65 (m, 1H), 2.61 - 2.50 (m, 1H), 2.37 - 2.22 (m, 2H), 1.91 - 1.47 (m, 25H), 1.47 - 1.18 (m, 7H), 1.03 (d, J = 1.6 Hz, 3H), 0.97 (d, J = 1.5 Hz, 3H). ESI-MS m/z calc. 769.415, found 769.75 (M+l)+; Retention time: 3.61 minutes using Method A.

[0327] The second eluting peak is lyophilised to afford (2S,3S,4S,5R,6R)-6-(((ls,4S)-4-(4-(7-(tert- butyl)-5-(4,4- dimethylcyclohexyl)furo[3,2-b]pyridine-2-carbonyl)-3,3-dimethylpiperazine-l- carbonyl)-l-methylcyclohexane-l-carbonyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2- carboxylic acid (11 mg). ^XH NMR (400 M Hz, Methanol-d4) δ 7.30 (d, J = 4.2 Hz, 1H), 7.21 (d, J = 5.1 Hz, 1H), 6.17 (d, J = 3.2 Hz, 1H), 4.07 - 3.94 (m, 2H), 3.88 - 3.80 (m, 1H), 3.76 (s, 1H), 3.72 (s, 1H), 3.70 - 3.44 (m, 5H), 2.81 - 2.66 (m, 1H), 2.63 - 2.49 (m, 1H), 2.30 (t, J = 11.7 Hz, 2H), 1.90 - 1.47 (m, 25H), 1.47 - 1.19 (m, 7H), 1.03 (d, J = 1.9 Hz, 3H), 0.97 (d, J = 1.6 Hz, 3H). ESI-MS m/z calc. 769.415, found 769.75 (M+l)⁺; Retention time: 3.59 minutes using Method A.

Example 1.26. 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-blpyridine-2-carbonyll-l,3,3- trimethyl-l,4-diazepan-2-one

Step I:

[0328] 2,6-lutidine (1.43 mL, 12.3 mmol) is added to a stirred solution of 2-(tert- butoxycarbonylamino)- 2-methyl-propanoic acid (500 mg, 2.46 mmol), 3-bromopropan-l-amine (HBr salt) (538.5 mg, 2.46 mmol) and HATU (1.403 g, 3.69 mmol) in DMF (7 mL). The reaction is stirred for 2 hours at RT. Water is added along with EtOAc and the phases are separated. The organic phase is washed twice with a water and brine mixture (1:1), dried over sodium sulfate, filtered and evaporated under reduced pressure. The crude is dissolved in EtOAc and is then washed with aqueous 1M HCI. The organic extract is washed with water, followed by brine, dried over sodium sulfate, filtered and evaporated under reduced pressure to give tert-butyl N- [2-(3-bromopropylamino)-l,l-dimethyl-2-oxo-ethyl]carbamate (900 mg, 113%). ¹H NM R (400 MHz, Chloroform-d) δ 6.68 (s, 1H), 4.82 (s, 1H), 3.47 (t, J = 6.5 Hz, 2H), 3.41 (q, J = 6.3 Hz, 2H), 2.09 (q, J = 6.4 Hz, 2H), 1.65 (d, J = 13.7 Hz, 1H), 1.47 (s, 6H), 1.44 (s, 9H). ESI-MS m/z calc. 322.0892, found 323.08 (M+l)+; Retention time: 0.58 minutes using Method J.

Step II: 2-amino-N-(3-bromopropyl)-2-methyl-propanamide

[0329] TFA (358 μί, 4.64 mmol) is added to a stirred solution of tert-butyl N-[2-(3- bromopropylamino)- l,l-dimethyl-2-oxo-ethyl]carbamate (150 mg, 0.464 mmol) in DCM (1.0 mL) at rt. The reaction is stirred for 4 hours at rt and is then concentrated under vacuum. Saturated NaHC0₃ solution and DCM are added and the mixture is stirred for 5 minutes. Phases are separated using a phase separator cartridge. The filtrate is concentrated to give 2-amino-N- (3-bromopropyl)-2-methyl-propanamide (105 mg, 101%) ESI-MS m/z calc. 222.03677, found 223.05 (M+l)+; Retention time: 0.38 minutes using Method J.

Step III: 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3- dimethyl-l,4-diazepan-2-one

[0330] A solution of the material obtained above (100.0 mg, 0.448 mmol) salt, NEt₃ (625 μί, 4.48 mmol) in DMF (9 mL) is stirred at rt for 24h. The mixture is treated with 7-tert-butyl-5-(4-chloro- 3-fluoro-phenyl)furo[3,2-b]pyridine-2-carboxylic acid (Intermediate A) (155.9 mg, 0.448 mmol) and HATU (255.6 mg, 0.672 mmol) added and the reaction is stirred at rt for the weekend. Water is slowly added and the mixture is stirred for 5 min and filtered and washed with heptane then dried to give 185 mg of the crude product. The solid is dissolved in NMP and purified on a Gemini-NX 250-21 column (Isco Combiflash EZ PREP) eluting with 45-80% CH₃CN (0.1% Formic Acid)/Water (0.1% Formic Acid) over 30 minutes. The desired fractions are combined and lyophilized to give 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]- 3,3-dimethyl-l,4-diazepan-2-one (125 mg, 57%). A portion of this material (65 mg) is purified on a Gemini-NX 250-21 column (Isco Combiflash EZ PREP) eluting with 40-70% CH₃CN (0.1% Formic Acid)/Water (0.1% Formic Acid) over 30 minutes. The desired fractions are combined and lyophilized to give 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]- 3,3-dimethyl-l,4-diazepan-2-one (30 mg). ^XH NMR (CDCI₃): Showed the presence of rotamers ca. 6:1 ratio. ^XH NMR (400 MHz, Chloroform-d) δ 9.02 (s, 1H), 7.84 (dd, J = 10.4, 2.0 Hz, 1H), 7.74 (dd, J = 8.4, 2.0 Hz, 1H), 7.57 (s, 1H), 7.49 (t, J = 7.9 Hz, 1H), 7.26 (s, 1H), 4.27 (t, J = 5.4 Hz, 2H), 3.50 (t, J = 5.9 Hz, 2H), 2.00 - 1.89 (m, 3H), 1.72 (s, 6H), 1.60 (s, 9H). ESI-MS m/z calc. 471.1725, found 472.63 (M+l)+; Retention time: 2.96 minutes using Method A.

Step IV: 4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-l,3,3- trimethyl-l,4-diazepan-2-one

[0331] NaH (9.0 mg of 60 %w/w, 0.22 mmol) is added to a stirred solution of 4-[7-tert-butyl-5-(4- chloro-3- fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-l,4-diazepan-2-one (55 mg, 0.11 mmol) in DMF (624 μί). After 15 minutes of stirring at rt, the solution is cooled to 0°C and then Mel (10.5 μί, 0.168 mmol) is added. The reaction is stirred for 2 h at RT. Another portion of NaH (6.7 mg of 60 %w/w, 0.17 mmol), followed by Mel (14 μί, 0.22 mmol) are added at 0°C. The reaction is stirred for 2 hours. Water is slowly added along with EtOAc and the phases are separated. Aqueous is extracted once more with EtOAc. The combined organic phases are washed with water and brine (1:1 mixture), then brine, dried over sodium sulfate, filtered and evaporated under reduced pressure. The crude residue is purified on reverse-phase PREP HPLC using CH3CN/ Water 0.1%FA to give upon lyophilization 4-[7-tert-butyl-5-(4-chloro- 3-fluoro-phenyl)furo[3,2-b]pyridine-2-carbonyl]-l,3,3-trimethyl-l,4-diazepan-2-one (29.40 mg, 53%) as a white solid. ESI-MS m/z calc. 485.18814, found 486.68 (M+l)+; Retention time: 3.02 minutes using Method A. ^XH NMR (400 M Hz, DMSO-d6) δ 8.17 (t, J = 11.0, 1.9 Hz, 1H), 8.04 (dd, J = 8.6, 2.0 Hz, 1H), 7.83 (s, 1H), 7.72 (t, J = 8.1 Hz, 1H), 7.52 (s, 1H), 4.06 (t, J = 5.4 Hz, 2H), 3.26 (t, J = 5.8 Hz, 2H), 3.17 (s, 3H), 1.73 (p, J = 5.7 Hz, 2H), 1.54 (s, 9H), 1.48 (s, 6H).

Example 1.27. Preparation of phosphonooxymethyl 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro- phenyl)furo[3,2-blpyridine-2-carbonyll-3,3-dimethyl-piperazin-l-yll-2-methyl-pyridine-3-carboxylate iH9)

Step I: ditert-butoxyphosphoryloxymethyl 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro- phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-l-yl]-2-methyl-pyridine-3- carboxylate 2] To a mixture of (110 mg, 0.120 mmol) and Kl (35.29 mg, 0.213 mmol) in DM F (1.9 mL) was added cesium carbonate (93.4 mg, 0.287 mmol). Then, ditert-butyl chloromethyl phosphate (54.0 mg, 0.209 mmol) was added drop-wise and the reaction mixture was stirred at room temperature for 3 hours and then was heated to 40°C for 90 min. A further portion of ditert- butyl chloromethyl phosphate (35.0 mg, 0.135 mmol) was added drop-wise and the mixture was stirred at 40°C for 90 min. The mixture was cooled to room temperature and water (25 mL) was added slowly. Then, the mixture was diluted with a 1:1 mixture (5 mL) of hexanes and EtOAc and the phases were separated. The aqueous layer was extracted twice using a 1:1 mixture of hexanes and EtOAc (2 X 5 mL). The combined organic layers were dried over MgS0₄, filtered and concentrated under reduced pressure. The obtained residue was purified by flash column chromatography on silica gel eluting with mixtures of hexanes and EtOAc affording ditert- butoxyphosphoryloxymethyl 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine- 2-carbonyl]-3,3-dimethyl-piperazin-l-yl]-2-methyl-pyridine-3-carboxylate (106.5 mg, 70%) as a yellow residue. ESI-MS m/z calc. 800.3117, found 801.43 (M+l)+; Retention time: 1.29 minutes using Method J. Step II: phosphonooxymethyl 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2- b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-l-yl]-2-methyl-pyridine-3-carboxylate

[0333] A suspension of di-tert-butoxyphosphoryloxymethyl 6-[4-[7-tert-butyl-5-(4-chloro-3-fluoro- phenyl)furo[3,2-b]pyridine-2-carbonyl]-3,3-dimethyl-piperazin-l-yl]-2-methyl-pyridine-3- carboxylate (107 mg, 0.122 mmol) in acetone (2.0 mL) and water (1.0 mL) was heated at 60°C for 20 hours. The suspension was cooled to rt and water was added (0.5 mL). The mixture was lyophilized over-night. The obtained residue was taken up in DMSO (1.6 mL), filtered through a 0.2 μΜ filter disk and the product was purified by reverse phase chromatography eluting with mixtures of ACN and water (0.1 % formic acid) affording phosphonooxymethyl 6-[4-[7-tert- butyl-5-(4-chloro-3-fluoro-phenyl)furo[3,2-b]pyridine-2- carbonyl]-3,3-dimethyl-piperazin-l-yl]- 2-methyl-pyridine-3-carboxylate (4.10 mg, 5%) as a white solid. ESI-MS m/z calc. 688.1865, found 689.21 (M+l)+; Retention time: 3.43 minutes using Method A.

[0334] Using one or more of the foregoing procedures, the following compounds were prepared as indicated in Table 1 and characterized as shown in Table 2, below.

Table 1: Furo[3,2- ?]pyridine Compounds

- 152 -

Table 2: Analytical Data

LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 8.98- 8.40 (m, IH, br),

8.17 (dd, J = 10.9, 2.0 Hz, 2H), 8.07 - 8.00 (m, 2H), 7.83 (s,-2 630.15 A: 3.81

IH), 7.76 - 7.67 (m, IH), 7.51 (s, IH), 7.28 - 6.83 (m, IH), 3.96 - 3.77 (m, 2H), 3.32 (s, 4H), 1.53 (s, 9H), 1.46 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 8.68 - 8.63 (m, IH), 8.17

(dd, J = 10.9, 2.0 Hz, IH), 8.03 (dd, J = 8.6, 1.9 Hz, IH), 7.82-3 628.25 A: 3.58 (s, IH), 7.76 - 7.65 (m, 3H), 7.49 (s, IH), 7.37 - 6.75 (m, IH),

3.99 - 3.67 (m, 2H), 3.72 - 3.18 (m, 4H), 1.53 (s, 9H), 1.45 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 10.51 (s, IH, br), 8.17 (dd,

J = 11.0, 2.1 Hz, IH), 8.04 (dd, J = 8.5, 2.0 Hz, IH), 7.83 (s,-4 607.2 A: 4.02 IH), 7.72 (dd, J = 8.1 Hz, IH), 7.53 (s, IH), 3.95 - 3.90 (m,

2H), 3.60 (s, 3H), 3.32 (s, 3H), 1.54 (s, 9H), 1.50 (s, 6H), 1.11 - 0.90 (m, IH), 0.60 - 0.53 (m, 2H), 0.33 - 0.27 (m, 2H).

IH NMR (400 M Hz, DMSO-d6) δ 10.48 (s, IH), 8.17 (dd, J =

11.0, 2.1 Hz, IH), 8.07 - 8.00 (m, IH), 7.83 (s, IH), 7.76 --5 637.2 A: 3.80 7.67 (m, IH), 7.52 (s, IH), 4.01 - 3.87 (m, 4H), 3.82 (s, IH),

3.61 - 3.56 (m, 3H), 3.44 - 3.18 (m, 3H), 1.88 - 1.80 (m, 2H), 1.77 - 1.59 (m, 2H), 1.53 (s, 9H), 1.49 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 8.05 (s, IH), 7.43 (s, IH),

7.16 (s, IH), 6.89 (s, IH), 4.04 (t, J = 5.6 Hz, 2H), 3.88 (s,-6 561.3 A: 3.62 2H), 3.67 - 3.53 (m, 2H), 2.80 - 2.64 (m, IH), 2.29 (s, 3H),

1.83 - 1.65 (m, 4H), 1.51 (s, 6H), 1.47 (s, 11H), 1.35 (dt, J = 13.9, 6.9 Hz, 2H), 0.98 (s, 3H), 0.96 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.18 (dd, J = 11.0, 2.1 Hz,

IH), 8.10 - 7.99 (m, 2H), 7.84 (s, IH), 7.72 (t, J = 8.1 Hz,-7 579.1 A: 3.65 IH), 7.56 (s, IH), 6.96 (s, IH), 4.09 (t, J = 5.6 Hz, 2H), 3.91

(s, 2H), 3.62 (t, J = 5.7 Hz, 2H), 2.30 (s, 3H), 1.55 (s, 9H), 1.53 (s, 6H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 8.13 (d, J = 8.6 Hz, 2H), 8.03 (s, IH), 7.75 (s, IH), 7.62 - 7.45 (m, 3H), 6.89 (s, IH),-8 561.37 A: 3.46

4.05 (t, J = 5.6 Hz, 2H), 3.87 (s, 2H), 3.58 (t, J = 5.6 Hz, 2H), 2.26 (s, 3H), 1.51 (s, 9H), 1.49 (s, 6H). -9 689.21 A: 3.43

IH NMR (400 M Hz, DMSO-d6) δ 7.99 (d, J = 3.0 Hz, IH), 7.66 (dd, J = 8.5, 7.2 Hz, IH), 7.47 (d, J = 3.1 Hz, IH), 7.40 (s, IH), 7.24 (d, J = 7.1 Hz, IH), 7.13 (s, IH), 6.75 (d, J = 8.5-10 546.57 A: 4.19

Hz, IH), 4.13 - 3.89 (m, 4H), 3.60 (t, J = 5.6 Hz, 2H), 2.78 - 2.58 (m, IH), 1.82 - 1.61 (m, 4H), 1.56 - 1.38 (m, 17H), 1.31 (td, J = 13.2, 4.1 Hz, 2H), 0.95 (s, 3H), 0.92 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 11.16 (s, IH), 7.72 (dd, J = 8.6, 7.3 Hz, IH), 7.41 (s, IH), 7.29 (d, J = 7.2 Hz, IH), 7.13 (s, IH), 6.93 (d, J = 8.6 Hz, IH), 4.05 (t, J = 5.6 Hz, 2H), 3.96 (s, 2H), 3.66 (t, J = 5.6 Hz, 2H), 3.32 (s, 3H), 2.68 (tt, J = 11.5, 3.9 Hz, IH), 1.88 - 1.57 (m, 4H), 1.57 - 1.38 (m, 17H),-11 624.965 A: 4.53 1.31 (td, J = 13.1, 4.4 Hz, 2H), 0.95 (s, 3H), 0.92 (s, 3H).

[2], IH NMR (400 M Hz, DMSO-d6)□ 7.66 (t, J = 7.9 Hz, IH), 7.40 (s, IH), 7.25 (d, J = 7.3 Hz, IH), 7.13 (s, IH), 6.93 - 6.71 (m, IH), 4.04 (t, J = 5.6 Hz, 2H), 3.94 (s, 2H), 3.64 (s, 2H), 3.19 (s, 3H), 1.86 - 1.61 (m, 4H), 1.58 - 1.37 (m, 17H), 1.37 - 1.24 (m, 2H), 0.95 (s, 3H), 0.92 (s, 4H).

IH NMR (400 M Hz, DMSO-d6) δ 13.51 (s, IH, br), 10.73 (s, IH, br), 8.27 (s, IH, br), 8.17 (dd, J = 11.0, 2.0 Hz, IH), 8.07-12 617.25 A: 3.52 - 8.00 (m, IH), 7.83 (s, IH), 7.82 - 7.76 (m, IH, br), 7.72 (t, J

= 8.1 Hz, IH), 7.51 (s, IH), 3.94 - 3.80 (m, 2H), 3.60 - 3.42 (m, 4H), 1.53 (s, 9H), 1.46 (s, 6H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 8.17 (dd, J = 11.0, 2.1 Hz, IH), 8.04 (dd, J = 8.5, 2.0 Hz, IH), 7.84 (s, IH), 7.76 - 7.67-13 552.15 A: 4.49 (m, IH), 7.56 (s, IH), 3.88 - 3.80 (m, 2H), 3.55 - 3.47 (m,

2H), 3.44 (p, J = 6.8 Hz, IH), 3.37 (s, 2H), 1.53 (d, J = 2.1 Hz, 15H), 1.27 (s, 3H), 1.26 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.18 (dd, J = 11.0, 2.1 Hz, IH), 3.34 - 3.29 (m, 10H), 8.11 - 7.99 (m, IH), 7.85 (s, IH),-14 528.1 A: 3.96

7.76 - 7.69 (m, IH), 7.66 (s, IH), 4.00 (s, 2H), 3.93 (s, 2H), 1.82 (s, 6H), 1.53 (s, 9H).

IH NMR (400 M Hz, DMSO-d6) δ 12.44 (s, IH), 8.20 - 8.11 (m, 2H), 7.79 (s, IH), 7.58 (s,lH), 7.58 - 7.51 (m, 2H), 3.81-15 540.34 A: 3.95

(d, J = 5.2 Hz, 2H), 3.63 (s, 2H), 3.57 (t, J = 4.8 Hz, 2H), 1.72 (s, 6H), 1.52 (s, 9H), 1.12 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 12.46 (s, IH), 7.47 (s, IH), 7.17 (s, IH), 3.78 (t, J = 4.7 Hz, 2H), 3.62 (s, 2H), 3.55 (t, J = 4.8 Hz, 2H), 2.71 (tt, J = 11.9, 4.0 Hz, IH), 1.87 - 1.73 (m,-16 540.24 A: 3.80

2H), 1.70 (s, 7H), 1.67 (d, J = 3.2 Hz, IH), 1.49 (s, IH), 1.45 (s, 9H), 1.34 (td, J = 13.2, 4.2 Hz, 2H), 1.11 (s, 6H), 0.98 (s, 3H), 0.95 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.17 (dd, J = 11.0, 2.1 Hz, IH), 8.04 (dd, J = 8.4, 2.0 Hz, IH), 7.84 (s, IH), 7.72 (t, J = 8.1 Hz, IH), 7.58 (s, IH), 3.82 - 3.75 (m, 2H), 3.71 (s, 2H),-17 584.05 A: 4.19

3.55 (t, J = 4.8 Hz, 2H), 1.97 (dd, J = 11.9, 5.5 Hz, 2H), 1.70 (s, 6H), 1.66 - 1.56 (m, 4H), 1.52 (s, 9H), 2H obscured by tBu.

IH NMR (400 M Hz, DMSO-d6) δ 8.19 - 8.11 (m, 2H), 7.79 (s, IH), 7.60 - 7.53 (m, 3H), 3.84 - 3.75 (m, 2H), 3.71 (s, 2H),-18 566.2 A: 4.08

3.55 (t, J = 4.9 Hz, 2H), 2.04 - 1.92 (m, 2H), 1.70 (s, 6H), 1.66 - 1.55 (m, 4H), 1.52 (s, 9H), 1.49 (s, 2H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 7.45 (s, IH), 7.16 (s, IH),

3.75 (t, J = 4.6 Hz, 2H), 3.70 (s, 2H), 3.53 (t, J = 4.9 Hz, 2H),

2.76 - 2.66 (m, IH), 2.04 - 1.94 (m, 2H), 1.86 - 1.72 (m, 2H),-19 566.78 A: 4.22

1.68 (s, 6H), 1.65 - 1.54 (m, 4H), 1.50 (d, J = 12.3 Hz, 3H), 1.45 (s, 11H), 1.33 (td, J = 13.2, 4.1 Hz, 3H), 0.98 (s, 3H), 0.95 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.13 (dd, J = 8.9, 2.2 Hz, 2H), 7.75 (s, IH), 7.62 (dd, J = 8.5, 7.3 Hz, IH), 7.56 - 7.49 (m, 3H), 7.23 (d, J = 7.2 Hz, IH), 6.77 (d, J = 8.5 Hz, IH), 4.07 (t, J = 5.6 Hz, 2H), 3.95 (s, 2H), 3.65 (t, J = 5.7 Hz, 2H), 3.10 (s, 3H), 1.60 - 1.44 (m, 15H).

-20 625.01 C: 3.28

[2], IH NMR (400 M Hz, DMSO-d6) δ 11.17 (s, IH), 8.17 - 8.07 (m, 2H), 7.75 (s, IH), 7.71 (t, J = 8.0 Hz, IH), 7.59 - 7.50 (m, 3H), 7.29 (d, J = 7.2 Hz, IH), 6.93 (d, J = 8.6 Hz, IH), 4.08 (t, J = 5.6 Hz, 2H), 3.98 (s, 2H), 3.75 - 3.61 (m, 2H), 3.31 (s, 3H), 1.60 - 1.40 (m, 15H).

IH NMR (400 M Hz, DMSO-d6) δ 8.19 - 8.09 (m, 2H), 8.00 (s, IH), 7.75 (s, IH), 7.66 (dd, J = 8.5, 7.3 Hz, IH), 7.56 --21 546.66 A: 4.07 7.50 (m, 3H), 7.48 (s, IH), 7.24 (dd, J = 7.3, 0.6 Hz, IH),

6.76 (d, J = 8.5 Hz, IH), 4.23 - 3.81 (m, 4H), 3.62 (t, J = 5.5 Hz, 2H), 1.69 - 1.28 (m, 15H).

IH NMR (400 M Hz, Methanol-d4) δ 7.30 (d, J = 3.9 Hz, IH), 7.21 (d, J = 3.8 Hz, IH), 4.00 (dt, J = 15.4, 5.8 Hz, 2H), 3.80 (t, J = 5.7 Hz, IH), 3.71 (d, J = 8.4 Hz, 2H), 3.60 (t, J = 5.8 Hz,-22 594.3 A: 4.40

IH), 2.81 - 2.64 (m, 3H), 1.98 (d, J = 12.1 Hz, 2H), 1.88 - 1.69 (m, 4H), 1.66 - 1.34 (m, 27H), 1.03 (s, 3H), 0.97 (d, J = 1.3 Hz, 3H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, Methanol-d4) δ 8.08 - 7.87 (m, 2H), 7.72 (d, J = 4.1 Hz, IH), 7.54 - 7.45 (m, 2H), 7.40 (d, J = 5.5 Hz, IH), 4.03 (dt, J = 16.1, 5.8 Hz, 2H), 3.81 (t, J = 5.7 Hz,-23 594.25 A: 4.20

IH), 3.72 (d, J = 8.5 Hz, 2H), 3.66 - 3.57 (m, IH), 2.76 (s, IH), 2.70 (s, IH), 1.99 (d, J = 11.4 Hz, 2H), 1.67 - 1.44 (m, 23H).

IH NMR (400 M Hz, DMSO-d6) δ 10.89 (s, IH), 8.15 (dd, J = 11.0, 2.1 Hz, IH), 8.05 - 7.98 (m, IH), 7.81 (s, IH), 7.75 --24 671.57 C: 2.28 7.64 (m, 2H), 7.53 (s, IH), 7.27 (d, J = 7.2 Hz, IH), 6.96 (d, J

= 8.6 Hz, IH), 4.08 (t, J = 5.6 Hz, 2H), 3.96 (s, 2H), 3.70 (t, J = 5.6 Hz, 3H), 2.86 (s, 6H), 1.64 - 1.41 (m, 15H).

IH NMR (400 M Hz, DMSO-d6) δ 8.04 (s, IH), 7.54 (s, IH), 7.19 (s, IH), 4.12 (s, 3H), 4.11 - 4.06 (m, 2H), 4.01 (t, J = 4.6-25 522.11 A: 4.53 Hz, 2H), 2.71 (ddt, J = 11.8, 7.9, 3.9 Hz, IH), 1.81 (s, 6H),

1.78 - 1.63 (m, 4H), 1.49 (s, 2H), 1.47 (s, 9H), 1.34 (td, J = 13.2, 4.1 Hz, 2H), 0.98 (s, 3H), 0.96 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 11.98 (s, IH), 8.83 (s, 2H), 8.15 (dd, J = 11.0, 2.1 Hz, IH), 8.05 - 7.96 (m, IH), 7.81 (s,-26 643.61 C: 2.04

IH), 7.75 - 7.62 (m, IH), 7.53 (s, IH), 4.27 - 3.92 (m, 4H), 3.78 (t, J = 5.7 Hz, 2H), 3.23 (s, 3H), 1.73 - 1.31 (m, 15H).

IH NMR (400 M Hz, DMSO-d6) δ 8.79 (s, 2H), 8.15 (dd, J = 11.0, 2.1 Hz, IH), 8.01 (ddd, J = 8.5, 2.1, 0.7 Hz, IH), 7.92 --27 566.55 C: 1.95 7.75 (m, 2H), 7.74 - 7.61 (m, IH), 7.52 (s, IH), 7.28 (s, IH),

4.18 - 3.90 (m, 3H), 3.76 (t, J = 5.7 Hz, 2H), 1.61 - 1.38 (m, 15H).

IH NMR (400 M Hz, DMSO-d6) δ 8.25 - 8.09 (m, 2H), 8.05 --28 610.3 A: 4.00 7.95 (m, IH), 7.82 (s, IH), 7.75 - 7.65 (m, 2H), 7.61 (s, IH),

4.08 - 3.83 (m, 4H), 1.77 (s, 6H), 1.70 (s, 6H), 1.52 (s, 9H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 12.73 (s, IH), 8.14 (s, IH), 8.01 (t, J = 9.0 Hz, 2H), 7.80 (s, IH), 7.67 (t, J = 8.1 Hz, IH),-29 599.2 A: 4.35

7.52 (s, IH), 6.67 (s, IH), 4.08 (t, J = 5.6 Hz, 2H), 3.90 (s, 2H), 3.64 (t, J = 5.5 Hz, 2H), 1.51 (s, 15H).

IH NMR (400 M Hz, DMSO-d6) δ 13.08 (s, IH), 8.13 (dd, J = 11.0, 2.1 Hz, IH), 8.00 (dd, J = 8.5, 2.0 Hz, IH), 7.93 (s, IH),-30 599.2 A: 4.63 7.79 (s, IH), 7.68 (t, J = 8.1 Hz, IH), 7.51 (s, IH), 6.78 (d, J =

7.9 Hz, IH), 4.02 (t, J = 5.4 Hz, 2H), 3.62 (s, 2H), 3.48 (t, J = 5.5 Hz, 2H), 1.52 (s, 6H), 1.50 (s, 9H).

IH NMR (400 M Hz, DMSO-d6) δ 13.23 (s, IH), 8.15 (dd, J = 11.0, 2.1 Hz, IH), 8.01 (dd, J = 8.5, 2.1 Hz, IH), 7.81 (s, IH),-31 579.25 A: 4.25 7.69 (t, J = 8.1 Hz, IH), 7.53 (s, IH), 6.88 (s, IH), 6.83 (s,

IH), 4.06 (t, J = 5.5 Hz, 2H), 3.90 (s, 2H), 3.62 (t, J = 5.6 Hz, 2H), 2.36 (s, 3H), 1.52 (s, 9H), 1.50 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 8.17 (d, J = 8.7 Hz, 2H), 8.05 (s, IH), 7.81 (s, IH), 7.65 (s, IH), 7.57 (d, J = 8.6 Hz,-32 522.62 A: 4.32

2H), 4.12 (s, 5H), 4.10 - 3.97 (m, 2H), 1.83 (s, 6H), 1.55 (s, 9H).

IH NMR (400 M Hz, DMSO-d6) δ 8.22 - 8.14 (m, IH), 8.08 - 8.01 (m, IH), 7.84 (s, IH), 7.77 - 7.68 (m, IH), 7.55 (s, IH),-33 551.7 A: 5.18 7.46 (t, J = 7.9 Hz, IH), 6.13 (d, J = 8.1 Hz, IH), 6.01 (d, J =

7.8 Hz, IH), 4.10 - 4.02 (m, 2H), 3.90 (s, 2H), 3.80 (s, 3H), 3.64 - 3.56 (m, 2H), 1.55 (s, 9H), 1.53 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 10.91 (s, IH, br), 7.91 (d, J = 7.7 Hz, 2H), 7.69 - 7.53 (m, 3H), 7.39 (s, IH), 7.15 (s, IH),-34 610.24 A: 4.14 3.89 - 3.83 (m, 2H), 3.73 - 3.35 (m, 4H), 2.76 - 2.64 (m, IH),

1.81 - 1.62 (m, 4H), 1.51 - 1.40 (m, 17H), 1.39 - 1.28 (m, 2H), 0.97 (s, 3H), 0.95 (s, 3H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 11.97 (s, IH), 8.83 (s, 2H), 7.40 (s, IH), 7.13 (s, IH), 4.12 - 3.94 (m, 4H), 3.76 (t, J = 5.7-35 622.65 C: 2.06 Hz, 2H), 3.25 (s, 3H), 2.74 - 2.61 (m, IH), 1.82 - 1.62 (m,

4H), 1.62 - 1.36 (m, 17H), 1.37 - 1.25 (m, 2H), 0.95 (s, 3H), 0.92 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.14 (dd, J = 11.0, 2.1 Hz, IH), 8.08 (d, J = 9.2 Hz, IH), 8.01 (dd, J = 8.4, 2.0 Hz, IH),-36 590.2 A: 4.23

7.81 (s, IH), 7.69 (t, J = 8.1 Hz, IH), 7.53 (s, IH), 7.01 (s, IH), 4.10 (s, 2H), 3.95 (s, 2H), 3.69 (s, 2H), 1.52 (s, 15H).

IH NMR (400 M Hz, DMSO-d6) δ 13.34 (s, IH), 8.08 (d, J = 9.2 Hz, IH), 7.41 (s, IH), 7.13 (s, IH), 7.01 (s, IH), 4.06 (d, J-37 572.35 A: 4.25 = 6.1 Hz, 2H), 3.93 (s, 2H), 3.67 (s, 2H), 2.68 (tt, J = 11.9,

4.0 Hz, IH), 1.82 - 1.60 (m, 4H), 1.50 (s, 6H), 1.44 (s, 11H), 1.30 (td, J = 13.0, 4.0 Hz, 2H), 0.95 (s, 3H), 0.92 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.21 (d, J = 8.7 Hz, IH), 8.15 (dd, J = 11.0, 2.1 Hz, IH), 8.01 (dd, J = 8.5, 2.0 Hz, IH),-38 593.25 A: 4.33 7.91 (d, J = 8.6 Hz, IH), 7.82 (s, IH), 7.69 (t, J = 8.1 Hz, IH),

7.62 (s, IH), 4.25 (dd, J = 6.8, 3.2 Hz, 2H), 4.14 - 3.96 (m, 2H), 2.69 (s, 3H), 1.82 (s, 6H), 1.52 (s, 9H).

IH NMR (400 M Hz, DMSO-d6) δ 8.18 (dd, J = 11.0, 2.0 Hz, IH), 8.05 (dd, J = 8.4, 2.0 Hz, IH), 7.91 - 7.82 (m, 2H), 7.72 (t, J = 8.2 Hz, IH), 7.57 (s, IH), 6.69 (d, J = 8.9 Hz, IH), 4.12 (t, J = 5.6 Hz, 2H), 4.00 (s, 2H), 3.70 (t, J = 5.6 Hz, 2H), 2.62 (s, 3H), 1.55 (s, 15H).

-39 603.3 A: 4.19

[2], IH NMR (400 M Hz, DMSO-d6) δ 8.18 (dd, J = 11.0, 2.0 Hz, IH), 8.05 (dd, J = 8.4, 2.0 Hz, IH), 7.92 (d, J = 8.7 Hz, IH), 7.84 (s, IH), 7.72 (t, J = 8.1 Hz, IH), 7.56 (s, IH), 6.62 (d, J = 8.7 Hz, IH), 4.10 (t, J = 5.5 Hz, 2H), 3.97 (s, 2H), 3.68 (t, J = 5.6 Hz, 2H), 2.63 (s, 3H), 1.56 (s, 15H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 8.18 (dd, J = 11.0, 2.0 Hz, IH), 8.04 (dd, J = 8.4, 2.1 Hz, IH), 7.83 (d, J = 9.1 Hz, 2H),-40 560.25 A: 4.92 7.72 (t, J = 8.1 Hz, IH), 7.56 (s, IH), 6.62 (s, IH), 4.10 (d, J =

6.1 Hz, 2H), 3.99 (s, 2H), 3.69 (s, 2H), 1.54 (s, 9H), 1.53 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 11.38 (brs, IH), 8.15 (d, J = 8.5 Hz, 2H), 7.77 (s, IH), 7.59 - 7.46 (m, 3H), 4.01 - 3.89-41 A: 4.08

(m, 2H), 3.86 - 3.47 (m, 4H), 2.50 - 2.36 (m, IH), 2.31 - 2.09 (m, IH), 1.90 - 1.66 (m, 3H), 1.64 - 1.17 (m, 20H).

IH NMR (400 M Hz, DMSO-d6) δ 11.80 (brs, IH), 7.40 (d, J = 11.0 Hz, IH), 7.15 (s, IH), 3.99 - 3.83 (m, 2H), 3.83 - 3.47-42 A: 4.22 (m, 4H), 2.77 - 2.63 (m, IH), 2.48 - 2.36 (m, IH), 2.30 - 2.09

(m, IH), 1.90 - 1.63 (m, 7H), 1.60 - 1.18 (m, 24H), 0.97 (s, 3H), 0.95 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 10.47 (s, IH), 7.41 (s, IH), 7.16 (s, IH), 3.94 - 3.87 (m, 2H), 3.60 - 3.55 (m, 4H), 3.25-43 546.75 A: 3.69

(s, 3H), 2.77 - 2.65 (m, IH), 1.82 - 1.62 (m, 4H), 1.54 - 1.40 (m, 17H), 1.40 - 1.28 (m, 2H), 0.98 (s, 3H), 0.95 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 11.98 (s, IH), 8.84 (s, 2H), 8.20 - 8.06 (m, 2H), 7.75 (s, IH), 7.56 - 7.52 (m, 2H), 7.52-44 626.39 C: 2.00

(s, IH), 4.17 - 3.96 (m, 4H), 3.83 - 3.67 (m, 2H), 3.27 (s, 3H), 1.55 - 1.47 (m, 15H).

IH NMR (400 M Hz, DMSO-d6) δ 8.11 (dd, J = 11.0, 2.0 Hz, IH), 7.97 (dd, J = 8.4, 1.9 Hz, IH), 7.77 (s, IH), 7.65 (t, J = 8.1 Hz, IH), 7.48 (d, J = 7.9 Hz, IH), 3.96 - 3.82 (m, 2H),-45 573.55 A: 2.78

3.82 - 3.73 (m, IH), 3.71 - 3.50 (m, 3H), 3.46 - 3.36 (m, IH), 2.30 (dt, J = 24.5, 6.8 Hz, 2H), 1.71 (broad s, IH), 1.52 - 1.39 (m, 16H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 8.14 (dd, J = 11.0, 2.1 Hz,

IH), 8.07 - 7.94 (m, 2H), 7.80 (s, IH), 7.68 (t, J = 8.1 Hz,-46 590.2 A: 4.33 IH), 7.51 (s, IH), 7.31 (d, J = 7.6 Hz, IH), 4.03 (t, J = 5.3 Hz,

2H), 3.65 (s, 2H), 3.50 (t, J = 5.4 Hz, 2H), 1.52 (s, 6H), 1.50 (s, 9H).

IH NMR (400 M Hz, DMSO-d6) δ 8.21 (s, IH), 8.15 (dd, J =

11.0, 2.1 Hz, IH), 8.01 (dd, J = 8.4, 2.0 Hz, IH), 7.81 (s, IH),-47 604.27 A: 3.25 7.69 (t, J = 8.2 Hz, IH), 7.58 (s, IH), 7.35 (d, J = 4.1 Hz, 2H),

6.58 (t, J = 4.1 Hz, IH), 3.91 (d, J = 5.5 Hz, 2H), 3.35 (d, J = 5.8 Hz, 2H), 3.19 (s, 2H), 1.62 (s, 6H), 1.50 (s, 9H).

The compound appears to show rotamers as indicated.

One proton is not accounted for and is possibly hidden under solvent or another peak). IH NM R (400 MHz,

DMSO-d6) δ 11.73 (s, IH), 7.43 , 7.39 (s, IH), 7.16, 7.15 (s,

IH), 3.93 (t, J = 5.6 Hz, IH), 3.86 (t, J = 5.7 Hz, IH), 3.68 (t, J-48 580.46 A: 4.15

= 5.6 Hz, IH), 3.62 (s, IH), 3.61 (s, IH), 3.45 (t, J = 5.7 Hz, IH), 2.76 - 2.65 (m, 3H), 2.05 (dt, J = 12.9, 6.3 Hz, 2H), 1.84 - 1.65 (m, 4H), 1.59 (t, J = 5.2 Hz, 4H), 1.50 (d, J = 9.6 Hz, 5H), 1.45 (d, J = 3.3 Hz, 13H), 1.33 (td, J = 13.3, 4.1 Hz, 2H), 0.98 (s, 3H), 0.95 (s, 3H).

LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 12.25 (s, IH), 8.18 (dd, J = 11.0, 1.7 Hz, IH), 8.05 (d, J = 8.5 Hz, IH), 7.84 (d, J = 1.8 Hz, IH), 7.73 (td, J = 8.2, 1.6 Hz, IH), 7.55 (d, J = 11.2 Hz, IH), 3.97 (t, J = 5.8 Hz, IH), 3.92 (t, J = 5.8 Hz, IH), 3.80 (q, J = 7.1, 6.7 Hz, 2H), 3.73 (t, J = 5.9 Hz, IH), 3.65 (dd, J = 10.0, 5.3 Hz, 2H), 3.49 (q, J = 4.8 Hz, IH), 2.98 - 2.87 (m, IH), 2.84 (s, IH), 2.31 - 2.20 (m, IH), 2.03 - 1.82 (m, 3H), 1.54 (two singlets, 12H), 1.49 (s, 3H).

-49 600.57 A: 3.71

[2], IH NMR (400 M Hz, DMSO-d6) δ 12.24 (s, IH), 8.17 (dd, J = 11.0, 2.0 Hz, IH), 8.04 (d, J = 8.5 Hz, IH), 7.83 (d, J = 1.8 Hz, IH), 7.72 (td, J = 8.2, 1.6 Hz, IH), 7.53 (d, J = 11.2 Hz, IH), 3.96 (d, J = 5.6 Hz, IH), 3.92 (dd, J = 12.5, 6.8 Hz, IH), 3.76 (ddp, J = 19.9, 13.7, 7.4, 6.9 Hz, 3H), 3.68 - 3.57 (m, 2H), 3.48 (q, J = 4.9 Hz, IH), 2.89 (d, J = 10.5 Hz, IH), 2.83 (s, IH), 2.30 - 2.19 (m, IH), 2.01 - 1.80 (m, 3H), 1.53 (m, 12H), 1.48 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.20 - 8.11 (m, 2.2H), 7.78 (s, 0.6H), 7.76 (s, 0.4H), 7.60 - 7.52 (m, 2.6H), 7.50 (s, 0.4H), 6.53 (s, 0.4H), 3.96 (s, IH), 3.88 (t, J = 5.7 Hz, 0.9H), 3.85 - 3.72 (m, 1.3H), 3.69 (d, J = 5.5 Hz, 0.7H), 3.56 (s,-50 592.37 A: 3.69

1.1H), 3.09 - 2.93 (m, 1.2H), 2.79 - 2.70 (m, 0.8H), 2.01 - 1.92 (m, IH), 1.54 (s, 6.4H), 1.51 (s, 5.2H), 1.45 (s, 1.6H), 1.33 (s, 1.6H), 1.23 (s, 0.4H), 0.85 (t, J = 7.0 Hz, 3H), 0.81 - 0.75 (m, 3H).

IH NMR (400 M Hz, Chloroform-d) δ 7.98 - 7.92 (m, 2H), 7.64 - 7.58 (m, 2H), 7.49 - 7.43 (m, 3H), 6.31 (d, J = 8.8 Hz,-51 542.26 A: 4.74

IH), 4.12 (t, J = 5.7 Hz, 2H), 3.99 (s, 2H), 3.72 (t, J = 5.6 Hz, 2H), 2.60 (s, 3H), 1.65 (s, 6H), 1.57 (s, 9H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 12.61 (s, IH), 8.18 (dd, J =

11.0, 2.1 Hz, IH), 8.15 - 8.07 (m, IH), 8.04 (dd, J = 8.4, 2.0-52 583.44 A: 4.25 Hz, IH), 7.84 (s, IH), 7.72 (t, J = 8.1 Hz, IH), 7.56 (s, IH),

6.62 (s, IH), 4.12 (t, J = 5.6 Hz, 2H), 3.93 (s, 2H), 3.68 (t, J = 5.7 Hz, 2H), 1.55 (s, 9H), 1.54 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 12.92 (s, IH), 8.21 - 8.11

(m, 2H), 8.04 (dd, J = 8.5, 2.0 Hz, IH), 7.83 (s, IH), 7.71 (t, J-53 583.68 A: 4.41 = 8.1 Hz, IH), 7.55 (s, IH), 6.45 (dd, J = 8.2, 3.1 Hz, IH),

4.14 - 4.00 (m, 2H), 3.63 (s, 2H), 3.52 (dd, J = 6.6, 4.3 Hz, 2H), 1.55 (s, 6H), 1.53 (s, 9H).

IH NMR (400 M Hz, DMSO-d6) δ 8.14 (dd, J = 11.0, 2.1 Hz,

IH), 8.00 (dd, J = 8.5, 2.0 Hz, IH), 7.80 (d, J = 0.9 Hz, IH),-54 573.5 A: 2.77 7.68 (t, J = 8.1 Hz, IH), 7.51 (d, J = 7.6 Hz, IH), 3.94 (dt, J =

11.8, 5.3 Hz, 2H), 3.71 (s, IH), 3.44 (d, J = 13.0 Hz, IH), 1.77 (s, IH), 1.58 (s, IH), 1.55 - 1.44 (m, 13H).

The NM R showed the presence of rotamers as indicated.

IH NMR (400 M Hz, DMSO-d6) δ 11.95 (s, IH), 8.17 (dd, J =

11.0, 2.4 Hz, IH), 8.03 (dd, J = 8.6, 2.3 Hz, IH), 7.84, 7.83

(s, IH), 7.72 (td, J = 8.2, 2.4 Hz, IH), 7.56, 7.51 (s, IH), 3.92-55 596.61 A: 3.81

(dt, J = 11.8, 5.6 Hz, 2H), 3.76 (t, J = 5.6 Hz, IH), 3.71 (s, IH), 3.64 (s, IH), 3.51 - 3.44 (m, IH), 2.91 - 2.65 (m, IH), 2.11 - 1.90 (m, 4H), 1.78 (d, J = 3.6 Hz, 2H), 1.64 (d, J = 3.0 Hz, IH), 1.54 (s, 3H), 1.53, 1.52 (s, 9H), 1.46 (s, 3H).

LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

The IH NM showed the presence of rotamers as indicated. IH NMR (400 MHz, DMSO-d6) δ 12.13 (s, IH), 8.20 - 8.11 (m, 2H), 7.79, 7.78 (s, IH), 7.58 - 7.56 (m, 2H), 7.55, 7.52 (s, IH), 3.97 (t, J = 5.8 Hz, IH), 3.91 (d, J = 6.1 Hz,-56 582.15 A: 3.67

IH), 3.86 - 3.69 (m, 3H), 3.69 - 3.57 (m, 2H), 3.49 (q, J = 4.6 Hz, IH), 3.00 - 2.87 (m, IH), 2.84 (s, IH), 2.27 (dq, J = 10.9, 6.0, 5.2 Hz, IH), 1.91 (ddt, J = 23.2, 12.6, 6.7 Hz, 3H), 1.54 (d, J = 3.0 Hz, 12H), 1.49 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.18 (dd, J = 11.0, 2.0 Hz, IH), 8.04 (dd, J = 8.4, 2.0 Hz, IH), 7.84 (s, IH), 7.77 - 7.69-57 551.26 B: 3.37 (m, 2H), 7.56 (s, IH), 7.22 - 7.15 (m, IH), 6.93 (dd, J = 7.6,

4.9 Hz, IH), 3.89 (s, 3H), 3.84 - 3.76 (m, 2H), 3.30 - 3.23 (m, 2H), 3.11 (s, 2H), 1.57 (s, 6H), 1.54 (s, 9H).

IH NMR (400 M Hz, DMSO-d6) δ 8.19 (dd, J = 10.9, 2.0 Hz, IH), 8.05 (dd, J = 8.5, 2.0 Hz, IH), 7.85 (s, IH), 7.78 - 7.70-58 569.26 A: 5.40 (m, IH), 7.66 (d, J = 2.5 Hz, IH), 7.57 (s, IH), 7.14 (dd, J =

10.3, 2.7 Hz, IH), 3.89 (s, 3H), 3.87 - 3.79 (m, 2H), 3.44 - 3.34 (m, 2H), 3.26 (s, 2H), 1.57 (s, 6H), 1.55 (s, 9H).

IH NMR (400 M Hz, Methanol-d4) δ 7.35 - 7.25 (m, IH), 7.26 - 7.15 (m, IH), 5.52 (d, J = 7.8 Hz, IH), 4.05 - 3.95 (m, 2H), 3.83 (t, J = 5.6 Hz, 2H), 3.76 (s, IH), 3.72 (s, IH), 3.63 --59 769.75 A: 3.61 3.56 (m, IH), 3.52 (broad s, IH), 3.47 - 3.40 (m, IH), 3.36

(t, J = 8.4 Hz, IH), 2.78 - 2.65 (m, IH), 2.61 - 2.50 (m, IH), 2.37 - 2.22 (m, 2H), 1.91 - 1.47 (m, 25H), 1.47 - 1.18 (m, 7H), 1.03 (d, J = 1.6 Hz, 3H), 0.97 (d, J = 1.5 Hz, 3H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, Methanol-d4) δ 7.30 (d, J = 4.2 Hz, IH), 7.21 (d, J = 5.1 Hz, IH), 6.17 (d, J = 3.2 Hz, IH), 4.07 - 3.94 (m, 2H), 3.88 - 3.80 (m, IH), 3.76 (s, IH), 3.72 (s, IH), 3.70 --60 769.75 A: 3.59

3.44 (m, 5H), 2.81 - 2.66 (m, IH), 2.63 - 2.49 (m, IH), 2.30 (t, J = 11.7 Hz, 2H), 1.90 - 1.47 (m, 25H), 1.47 - 1.19 (m, 7H), 1.03 (d, J = 1.9 Hz, 3H), 0.97 (d, J = 1.6 Hz, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.23 - 8.03 (m, 2H), 7.75 (s, IH), 7.60 - 7.43 (m, 4H), 6.78 (d, J = 8.7 Hz, IH), 4.04 (t,-61 561.54 C: 2.18

J = 5.6 Hz, 2H), 3.89 (s, 2H), 3.60 (t, J = 5.6 Hz, 2H), 2.30 (s, 3H), 1.60 - 1.29 (m, 15H).

IH NMR (400 M Hz, DMSO-d6) δ 8.16 (d, J = 8.6 Hz, 2H), 7.89 (d, J = 8.7 Hz, IH), 7.79 (s, IH), 7.58 (s, IH), 7.56 (d, J =-62 585.27 A: 4.21 1.2 Hz, 2H), 6.69 (d, J = 8.9 Hz, IH), 4.12 (t, J = 5.6 Hz, 2H),

4.00 (s, 2H), 3.71 (t, J = 5.6 Hz, 2H), 2.62 (s, 3H), 1.55 (s, 15H).

IH NMR (400 M Hz, Chloroform-d) δ 7.61 (d, J = 8.8 Hz, IH), 7.37 (s, IH), 7.08 (s, IH), 6.30 (d, J = 8.8 Hz, IH), 4.09-63 542.41 A: 5.04 (t, J = 5.7 Hz, 2H), 3.97 (s, 2H), 3.74 - 3.62 (m, 2H), 2.78 - 2.65 (m, IH), 2.59 (s, 3H), 1.84 - 1.75 (m, 4H), 1.63 (s, 6H), 1.51 (s, 11H), 1.43 - 1.34 (m, 2H), 1.02 (s, 3H), 0.98 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 7.89 (d, J = 8.8 Hz, IH), 7.44 (s, IH), 7.17 (s, IH), 6.67 (d, J = 8.9 Hz, IH), 4.08 (t, J = 5.6 Hz, 2H), 3.98 (s, 2H), 3.68 (t, J = 5.6 Hz, 2H), 2.76 - 2.67-64 585.42 A: 4.58

(m, IH), 2.61 (s, 3H), 1.84 - 1.66 (m, 4H), 1.53 (s, 6H), 1.48 (s, 11H), 1.39 - 1.29 (m, 2H), 1.23 (s, IH), 0.98 (s, 3H), 0.96 (s, 3H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 12.19 (s, IH), 7.42 (d, J = 12.9 Hz, IH), 7.16 (d, J = 1.7 Hz, IH), 3.93 (t, J = 5.7 Hz, IH), 3.88 (t, J = 5.7 Hz, IH), 3.79 (q, J = 7.0 Hz, 2H), 3.72 (d, J = 6.9 Hz, IH), 3.63 (dd, J = 9.5, 5.0 Hz, 2H), 3.47 (s, IH), 2.96-65 583.02 A: 3.68

- 2.85 (m, IH), 2.83 (s, IH), 2.76 - 2.64 (m, 2H), 2.26 (dd, J = 11.9, 5.9 Hz, IH), 1.90 (ddt, J = 24.9, 13.9, 7.8 Hz, 3H), 1.82 - 1.64 (m, 4H), 1.52 (s, 3H), 1.50 (s, IH), 1.46 (d, J = 3.3 Hz, 14H), 1.39 - 1.27 (m, 2H), 0.98 (s, 3H), 0.96, (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.15 (dd, J = 11.0, 2.1 Hz, IH), 8.07 - 7.97 (m, IH), 7.82 (s, IH), 7.72 - 7.66 (m, IH),-66 554.64 A: 4.29

7.63 (s, IH), 4.02 (s, 5H), 3.92 (d, J = 5.2 Hz, 2H), 2.04 (s, 3H), 1.80 (s, 6H), 1.50 (s, 9H).

IH NMR (400 M Hz, DMSO-d6) δ 12.43 (s, IH), 7.49 (d, J = 8.6 Hz, IH), 7.40 (s, IH), 7.13 (s, IH), 6.76 (d, J = 8.7 Hz, IH), 4.01 (t, J = 5.5 Hz, 2H), 3.87 (s, 2H), 3.58 (t, J = 5.6 Hz,-67 561.46 C: 2.25

2H), 2.74 - 2.62 (m, IH), 2.30 (s, 3H), 1.80 - 1.62 (m, 4H), 1.56 - 1.37 (m, 17H), 1.37 - 1.25 (m, 2H), 0.95 (s, 3H), 0.92 (s, 3H).

IH NMR (400 M Hz, Chloroform-d) δ 9.02 (s, IH), 7.84 (dd, J = 10.4, 2.0 Hz, IH), 7.74 (dd, J = 8.4, 2.0 Hz, IH), 7.57 (s,-68 472.63 A: 2.96 IH), 7.49 (t, J = 7.9 Hz, IH), 7.26 (s, IH), 4.27 (t, J = 5.4 Hz,

2H), 3.50 (t, J = 5.9 Hz, 2H), 2.00 - 1.89 (m, 3H), 1.72 (s, 6H), 1.60 (s, 9H).

IH NMR (400 M Hz, DMSO-d6) δ 11.42 (s, IH, br), 8.18 (dd, J = 11.0, 2.0 Hz, IH), 8.08 - 8.00 (m, IH), 7.84 (s, IH), 7.76 --69 537.54 A: 4.14 7.67 (m, IH), 7.55 (s, IH), 6.96 - 6.91 (m, IH), 6.68 - 6.61

(m, IH), 6.14 (t, J = 6.8 Hz, IH), 3.85 - 3.78 (m, 2H), 3.32 - 3.26 (m, 4H), 1.54 (s, 15H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 11.20 (s, IH), 8.19 - 8.07 (m, 2H), 7.75 (s, IH), 7.57 - 7.45 (m, 4H), 7.04 - 6.60 (m,-70 638.66 A: 4.60

IH), 4.05 (t, J = 5.4 Hz, 2H), 3.84 (s, 2H), 3.65 (s, 2H), 2.31 (s, 3H), 1.51 (s, 15H).

IH NMR (400 M Hz, DMSO-d6) δ 8.17 (t, J = 11.0, 1.9 Hz, IH), 8.04 (dd, J = 8.6, 2.0 Hz, IH), 7.83 (s, IH), 7.72 (t, J =-71 486.68 A: 3.02 8.1 Hz, IH), 7.52 (s, IH), 4.06 (t, J = 5.4 Hz, 2H), 3.26 (t, J =

5.8 Hz, 2H), 3.17 (s, 3H), 1.73 (p, J = 5.7 Hz, 2H), 1.54 (s, 9H), 1.48 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 11.31 (s, IH), 8.17 (dd, J = 11.0, 2.0 Hz, IH), 8.08 - 8.00 (m, IH), 7.84 (s, IH), 7.76 --72 555.76 A: 4.20 7.67 (m, IH), 7.55 (s, IH), 7.00 - 6.94 (m, IH), 6.65 - 6.57

(m, IH), 3.92 - 3.85 (m, 2H), 3.62 (s, 2H), 3.45 - 3.38 (m, 2H), 1.54 (s, 9H), 1.52 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 11.39 (s, IH), 7.37 (s, IH), 7.12 (s, IH), 3.91 (t, J = 5.2 Hz, IH), 3.84 (t, J = 5.8 Hz, IH), 3.64 (t, J = 5.5 Hz, IH), 3.62 - 3.57 (m, 2H), 3.45 (t, J = 5.7-73 646.13 A: 3.94 Hz, IH), 3.09 (s, 3H), 2.75 - 2.60 (m, IH), 2.18 (dt, J = 16.9,

8.3 Hz, 2H), 1.88 - 1.54 (m, 6H), 1.48 (s, 3H), 1.47 - 1.38 (m, 14H), 1.30 (td, J = 13.4, 4.3 Hz, 2H), 1.07 (s, 6H), 0.94 (s, 3H), 0.92 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.21 - 8.13 (m, IH), 8.07 - 8.00 (m, IH), 7.83 (s, IH), 7.76 - 7.67 (m, IH), 7.53 (s, IH), 4.05 (q, J = 7.1 Hz, 2H), 3.91 - 3.84 (m, 2H), 3.77 - 3.69 (m,-74 626.67 A: 4.45

2H), 3.48 - 3.40 (m, 2H), 3.35 (d, J = 27.1 Hz, 6H), 2.04 - 1.99 (m, IH), 1.53 (s, 9H), 1.51 (s, 6H), 1.18 (t, J = 7.1 Hz, 3H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, Chloroform-d) δ 7.84 (dt, J = 10.5, 2.7 Hz, IH), 7.77 - 7.72 (m, IH), 7.58 (d, J = 4.7 Hz, IH), 7.50 (td, J = 7.9, 3.0 Hz, IH), 7.43 (s, IH), 6.05 (brs, IH), 5.28-75 573.5 A: 3.65 (brs, IH), 3.97 (t, J = 5.6 Hz, IH), 3.91 (t, J = 5.8 Hz, IH),

3.77 - 3.68 (m, 3H), 3.56 (s, IH), 2.65 (s, IH), 2.59 (s, IH), 1.66 (s, 3H), 1.61 (s, 3H), 1.57 (s, 4H), 1.55 (s, 5H), 1.38 (s, 6H).

IH NMR (400 M Hz, Chloroform-d) δ 7.83 (ddd, J = 10.4, 3.7, 2.0 Hz, IH), 7.73 (dt, J = 8.5, 2.7 Hz, IH), 7.57 (d, J = 4.3 Hz, IH), 7.49 (td, J = 8.0, 2.4 Hz, IH), 7.41 (s, IH), 6.24 (brs, IH), 5.46 (brs, IH), 3.96 (t, J = 5.5 Hz, IH), 3.90 (dd, J-76 613.56 A: 4.00

= 7.3, 4.3 Hz, IH), 3.76 (t, J = 5.6 Hz, IH), 3.73 - 3.66 (m, 2H), 3.57 (s, IH), 2.66 (s, IH), 2.60 (s, IH), 2.04 - 1.93 (m, 2H), 1.78 - 1.68 (m, 2H), 1.65 (s, 3H), 1.60 (s, 3H), 1.58 - 1.42 (m, 15H).

IH NMR (400 M Hz, DMSO-d6) δ 8.22 - 8.10 (m, IH), 8.01 (dd, J = 8.5, 2.3 Hz, IH), 7.80 (d, J = 3.7 Hz, IH), 7.76 - 7.60-77 675.49 A: 4.20 (m, IH), 7.50 (d, J = 10.2 Hz, IH), 4.02 - 3.90 (m, IH), 3.90 - 3.79 (m, IH), 3.75 - 3.52 (m, 3H), 3.51 - 3.36 (m, IH), 3.28 (s, 3H), 2.16 - 1.96 (m, 2H), 1.67 - 1.33 (m, 19H).

IH NMR (400 M Hz, DMSO-d6) δ 12.22 (s, IH), 8.20 - 8.12 (m, IH), 8.06 - 7.99 (m, IH), 7.82 (s, IH), 7.75 - 7.66 (m,-78 597.34 A: 3.71 IH), 7.53 (s, IH), 3.91 - 3.84 (m, 2H), 3.76 - 3.65 (m, 2H),

3.48 - 3.40 (m, 2H), 3.40 - 3.33 (m, 4H), 2.00 - 1.94 (m, 2H), 1.53 (s, 9H), 1.51 (s, 6H), 1.17 - 1.11 (m, IH).

IH NMR (400 M Hz, DMSO-d6) δ 10.01 (s, IH), 7.39 (s, IH), 7.16 (s, IH), 6.59 - 6.52 (m, IH), 3.84 - 3.79 (m, 2H), 3.21-79 561.73 A: 3.72 (s, 3H), 2.80 - 2.62 (m, IH), 2.00 (s, IH), 1.85 - 1.58 (m, 7H),

1.58 - 1.42 (m, 18H), 1.40 - 1.27 (m, 2H), 0.98 (s, 3H), 0.95 (s, 3H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 11.96 (brs, IH), 8.17 (d, J = 8.4 Hz, 2H), 7.84 - 7.78 (m, 2H), 7.60 - 7.54 (m, 3H), 7.01-80 562.33 A: 3.85 (brs, 2H), 5.94 (d, J = 8.8 Hz, IH), 4.08 (t, J = 5.6 Hz, 2H),

3.94 (s, 2H), 3.65 (t, J = 5.5 Hz, 2H), 1.55 (s, 9H), 1.53 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 11.96 (brs, IH), 7.81 (d, J = 8.7 Hz, IH), 7.43 (s, IH), 7.17 (s, IH), 7.02 (brs, 2H), 5.93 (d, J = 8.8 Hz, IH), 4.05 (t, J = 5.5 Hz, 2H), 3.93 (s, 2H), 3.64-81 562.43 A: 4.02

(t, J = 5.5 Hz, 2H), 2.77 - 2.67 (m, IH), 1.85 - 1.66 (m, 4H), 1.52 (s, 6H), 1.48 (s, 10H), 1.35 (td, J = 13.2, 4.0 Hz, 3H), 0.99 (s, 3H), 0.97 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.36 (s, IH), 7.39 (s, IH), 7.17 (s, IH), 3.86 - 3.75 (m, IH), 3.30 - 3.13 (m, 3H), 2.77 --82 440.23 A: 2.77 2.67 (m, IH), 2.00 - 1.95 (m, IH), 1.85 - 1.40 (m, 10H), 1.56

(s, 3H), 1.47 (s, 9H), 1.43 (s, 3H), 1.40 - 1.28 (m, 2H), 0.98 (s, 3H), 0.96 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 10.06 (s, IH, br), 8.18 (dd, J = 11.0, 2.0 Hz, IH), 8.04 (dd, J = 8.4, 2.0 Hz, IH), 7.84 (s, IH), 7.77 - 7.68 (m, IH), 7.55 (s, IH), 7.37 (t, J = 7.9 Hz,-83 537.33 A: 3.90

IH), 5.99 - 5.94 (m, IH), 5.87 - 5.80 (m, IH), 4.12 - 3.99 (m, 2H), 3.84 (s, 2H), 3.54 (t, J = 5.5 Hz, 2H), 1.55 (s, 9H), 1.52 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 8.18 (dd, J = 11.0, 1.9 Hz, IH), 8.08 - 8.00 (m, IH), 7.83 (s, IH), 7.76 - 7.67 (m, IH),-84 564.33 A: 5.30 7.54 (s, IH), 7.29 (t, J = 8.0 Hz, IH), 5.87 (d, J = 8.1 Hz, IH),

5.82 (d, J = 8.0 Hz, IH), 4.06 - 3.98 (m, 2H), 3.85 (s, 2H), 3.62 - 3.54 (m, 2H), 2.97 (s, 6H), 1.54 (s, 9H), 1.52 (s, 6H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, Methanol-d4) δ 8.19 (d, J = 9.0 Hz, IH), 7.94 (dd, J = 10.7, 2.0 Hz, IH), 7.84 (dd, J = 8.5, 2.0 Hz, IH), 7.76 (s, IH), 7.59 (t, J = 8.0 Hz, IH), 7.43 (s, IH), 6.50 (d, J =-85 755.58 A: 3.77

9.1 Hz, IH), 5.67 (d, J = 7.2 Hz, IH), 4.18 (t, J = 5.7 Hz, 2H), 4.08 (s, 2H), 3.92 (d, J = 9.3 Hz, IH), 3.75 (t, J = 5.7 Hz, 2H), 3.63 - 3.44 (m, 3H), 2.68 (s, 3H), 1.62 (s, 6H), 1.59 (s, 9H).

IH NMR (400 M Hz, Chloroform-d) δ 7.84 (d, J = 10.4 Hz, IH), 7.74 (d, J = 8.4 Hz, IH), 7.58 (d, J = 1.4 Hz, IH), 7.50 (t, J = 7.6 Hz, IH), 7.43 (d, J = 8.5 Hz, IH), 4.00 (t, J = 5.5 Hz,-86 553.33 A: 4.15

IH), 3.94 (t, J = 5.7 Hz, IH), 3.82 - 3.72 (m, 3H), 3.56 (s, IH), 2.62 (s, IH), 2.57 (s, IH), 1.67 (s, 3H), 1.64 (s, 3H), 1.56 (s, 4H), 1.56 (s, 5H), 1.54 (s, 6H).

IH NMR (400 M Hz, Chloroform-d) δ 7.84 (d, J = 10.3 Hz, IH), 7.74 (d, J = 8.4 Hz, IH), 7.58 (s, IH), 7.53 - 7.47 (m, IH), 7.44 (d, J = 7.5 Hz, IH), 3.99 (t, J = 5.5 Hz, IH), 3.94 (t,-87 593.34 A: 4.52 J = 5.8 Hz, IH), 3.80 - 3.75 (m, 3H), 3.58 (s, IH), 2.61 (s,

IH), 2.57 (s, IH), 2.20 (t, J = 10.8 Hz, 2H), 1.81 - 1.68 (m, 5H), 1.66 (s, 3H), 1.63 (s, 3H), 1.56 (s, 4H), 1.55 (s, 5H), 1.46 - 1.36 (m, 2H), 1.28 - 1.15 (m, IH).

IH NMR (400 M Hz, DMSO-d6) δ 12.38 (s, IH), 8.17 (dd, J = 11.0, 2.1 Hz, IH), 8.04 (ddd, J = 8.5, 2.1, 0.7 Hz, IH), 7.94 (d, J = 8.4 Hz, IH), 7.83 (s, IH), 7.75 - 7.69 (m, IH), 7.55 (s,-88 595.15 A: 4.66

IH), 6.17 (d, J = 8.4 Hz, IH), 4.04 (t, J = 5.5 Hz, 2H), 3.87 (s, 3H), 3.67 (s, 2H), 3.58 (t, J = 5.5 Hz, 2H), 1.54 (s, 6H), 1.53 (s, 9H).

IH NMR (400 M Hz, DMSO-d6) δ 8.18 (dd, J = 11.0, 2.1 Hz, IH), 8.07 - 8.02 (m, IH), 7.84 (s, IH), 7.77 - 7.69 (m, IH),-89 536.14 A: 3.47 7.54 (s, IH), 7.18 (t, J = 7.9 Hz, IH), 5.73 (dd, J = 14.2, 7.8

Hz, 2H), 5.52 (s, 2H), 4.03 (t, J = 5.6 Hz, 2H), 3.51 (t, J = 5.6 Hz, 2H), 1.54 (s, 9H), 1.51 (s, 6H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min) -90 570.37 A: 4.32

IH NMR (400 M Hz, DMSO-d6) δ 8.21 - 8.13 (m, IH), 8.07 - 7.99 (m, IH), 7.81 (s, IH), 7.76 - 7.67 (m, IH), 7.45 (s, IH),-91 559.69 A: 5.15

3.69 (s, IH), 3.56 (s, 2H), 3.51 - 3.43 (m, 2H), 3.29 (s, IH), 1.93 (s, 2H), 1.59 - 1.48 (m, 15H), 1.43 (s, 9H).

IH NMR (400 M Hz, DMSO-d6) δ 8.20 - 8.14 (m, IH), 8.04 (t, J = 7.6 Hz, IH), 7.84 (s, 0.5H), 7.82 (s, 0.5H), 7.76 - 7.68 (m, IH), 7.55 (s, 0.5H), 7.50 (s, 0.5H), 3.95 - 3.89 (m, IH), 3.87 - 3.81 (m, IH), 3.69 - 3.61 (m, IH), 3.57 (s, IH), 3.51-92 636.61 A: 4.12

(s, IH), 3.41 - 3.35 (m, IH), 2.84 (s, IH), 2.80 (s, IH), 2.26 - 2.16 (m, 2H), 1.83 - 1.72 (m, 2H), 1.60 - 1.56 (m, IH), 1.54 (s, 5H), 1.51 (s, 4H), 1.50 (s, 3H), 1.40 (s, 4H), 1.38 - 1.18 (m, 4H).

IH NMR (400 M Hz, DMSO-d6) δ 8.20 - 8.13 (m, IH), 8.06 - 7.99 (m, IH), 7.84 (s, 0.5H), 7.82 (s, 0.5H), 7.75 - 7.67 (m, IH), 7.56 (s, 0.5H), 7.51 (s, 0.5H), 3.97 (t, J = 5.6 Hz, IH),-93 596.39 A: 3.72

3.88 (t, J = 5.8 Hz, IH), 3.72 (t, J = 5.6 Hz, IH), 3.66 (s, IH), 3.55 (s, IH), 3.41 (t, J = 5.7 Hz, IH), 2.93 (s, IH), 2.89 (s, IH), 1.54 (s, 7H), 1.51 (s, 5H), 1.45 (s, 6H), 1.43 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.19 - 8.13 (m, IH), 8.03 (d, J = 8.4 Hz, IH), 7.83 (s, IH), 7.71 (t, J = 8.1 Hz, IH), 7.53 (s, 0.5H), 7.51 (s, 0.5H), 3.97 - 3.84 (m, 2H), 3.62 (m, 2H),-94 610.39 A: 3.73

3.57 (s, IH), 3.47 (t, J = 5.7 Hz, IH), 2.25 - 2.18 (m, IH), 2.18 - 2.10 (m, IH), 2.02 - 1.93 (m, 2H), 1.52 (s, 4H), 1.52 (s, 5H), 1.50 (s, 3H), 1.47 (s, 3H), 1.39 (s, 3H), 1.38 (s, 3H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 8.21 - 8.13 (m, IH), 8.04 (t, J = 8.1 Hz, IH), 7.84 (s, 0.5H), 7.82 (s, 0.5H), 7.72 (q, J = 7.9 Hz, IH), 7.56 (s, 0.5H), 7.51 (s, 0.5H), 3.97 (t, J = 5.6 Hz, IH), 3.87 (t, J = 5.8 Hz, IH), 3.74 (t, J = 5.7 Hz, IH), 3.66 (s,-95 608.39 A: 3.81

IH), 3.53 (s, IH), 3.40 (t, J = 5.8 Hz, IH), 3.22 (s, IH), 3.16 (s, IH), 2.48 - 2.41 (m, 2H), 2.25 (q, J = 10.0, 9.2 Hz, 2H), 2.12 - 2.02 (m, IH), 2.01 - 1.90 (m, IH), 1.55 (s, 8H), 1.51 (s, 4H), 1.43 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.22 - 8.13 (m, IH), 8.08 --96 540.18 A: 3.01 8.00 (m, IH), 7.84 (s, IH), 7.77 - 7.67 (m, IH), 7.57 (s, IH),

3.93 (s, 4H), 3.81 (s, 2H), 1.55 - 1.50 (m, 15H).

IH NMR (400 M Hz, DMSO-d6) δ 13.14 (brs, IH), 8.24 (d, J = 8.6 Hz, IH), 7.94 (d, J = 8.7 Hz, IH), 7.54 (s, IH), 7.18 (s,-97 575.93 A: 4.58 IH), 4.29 - 4.23 (m, 2H), 4.07 - 3.98 (m, 2H), 2.77 - 2.65 (m,

4H), 1.84 (s, 6H), 1.83 - 1.65 (m, 4H), 1.47 (s, 11H), 1.40 - 1.29 (m, 2H), 0.98 (s, 3H), 0.96 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 7.45 (s, IH), 7.17 (s, IH), 3.95 - 3.85 (m, 5H), 3.78 (s, 2H), 2.76 - 2.66 (m, IH), 1.86 --98 522.48 A: 3.05

1.62 (m, 4H), 1.50 (s, 6H), 1.46 (s, 10H), 1.39 - 1.27 (m, 2H), 0.98 (s, 3H), 0.95 (s, 3H).

IH NMR (400 M Hz, DMSO-d6) δ 8.31 (s, IH), 8.21 - 8.14 (m, IH), 8.07 - 8.00 (m, IH), 7.83 (s, IH), 7.76 - 7.67 (m, IH), 7.52 (s, IH), 6.52 (s, IH, broad), 4.10 (s, IH), 3.90 --99 554.36 A: 3.21

3.81 (m, IH), 3.52 - 3.47 (m, IH), 2.11 - 2.05 (m, IH), 1.91 - 1.67 (m, 3H), 1.57 (s, 3H), 1.54 (s, 9H), 1.46 (s, 3H). Some peaks obscured by broad water peak. LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 8.22 - 8.14 (m, IH), 8.07 - 8.00 (m, IH), 7.83 (s, IH), 7.76 - 7.67 (m, IH), 7.57 - 7.52 (m, IH), 6.52 (s, IH, br), 4.63 - 4.56 (m, IH), 3.94 - 3.83 (m,-100 568.46 A: 3.45

IH), 3.46 - 3.34 (m, IH), 3.18 (s, 3H), 2.16 - 2.05 (m, IH), 2.05 - 1.95 (m, 2H), 1.72 - 1.62 (m, IH), 1.59 (s, 3H), 1.54 (s, 9H), 1.49 (s, 3H). Peaks obscured by broad water peak.

IH NMR (400 M Hz, DMSO-d6) δ 9.99 (s, IH, br), 8.21 - 8.13 (m, IH), 8.07 - 8.00 (m, IH), 7.83 (s, IH), 7.76 - 7.67 (m,-101 579.71 A: 3.95 IH), 7.52 (s, IH), 6.57 - 6.50 (m, IH), 3.86 - 3.78 (m, 2H),

3.38 - 3.27 (m, IH), 3.19 (s, 3H), 2.02 (s, IH), 1.83 - 1.73 (m, IH), 1.55 (d, J = 14.2 Hz, 14H), 1.49 - 1.44 (m, 3H).

IH NMR (400 M Hz, Methanol-d4) δ 8.07 (d, J = 9.0 Hz, IH), 8.00 - 7.92 (m, IH), 7.90 - 7.82 (m, IH), 7.77 (s, IH), 7.66 - 7.57 (m, IH), 7.41 (s, IH), 6.76 (d, J = 9.0 Hz, IH), 4.30 (q, J-102 621.72 B: 4.09

= 7.1 Hz, 2H), 4.12 (s, 2H), 3.96 - 3.88 (m, 2H), 3.89 - 3.81 (m, 2H), 2.68 (s, 3H), 2.26 - 2.18 (m, 2H), 1.65 - 1.60 (m, 15H), 1.37 (t, J = 7.1 Hz, 3H). -103 580.6 A: 4.02

IH NMR (400 M Hz, DMSO-d6) δ 12.03 (s, IH), 8.18 (d, J = 11.3 Hz, IH), 8.04 (d, J = 8.5 Hz, IH), 7.84 (s, IH), 7.73 (t, J-104 572.65 A: 3.74 = 8.1 Hz, IH), 7.53 (d, J = 1.2 Hz, IH), 6.30 (s, IH), 3.91 (t, J

= 5.6 Hz, 2H), 3.53 (d, J = 4.5 Hz, 4H), 1.54 (s, 9H), 1.49 (s, 6H), 1.37 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 12.62 (s, IH), 8.18 (dd, J = 10.9, 1.9 Hz, IH), 8.04 (d, J = 8.4 Hz, IH), 7.84 (s, IH), 7.73-105 573.6 A: 4.15 (t, J = 8.1 Hz, IH), 7.54 (s, IH), 3.89 (d, J = 6.8 Hz, 2H), 3.60

(dd, J = 11.1, 5.2 Hz, 2H), 3.50 (d, J = 18.1 Hz, 2H), 1.54 (s, 9H), 1.53 (s, 3H), 1.50 (s, 9H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, Chloroform-d) δ 7.87 - 7.80 (m, IH), 7.76 - 7.69 (m, IH), 7.59 (s, IH), 7.55 - 7.44 (m, 2H), 5.84-106 539.28 A: 3.49

(s, 2H), 4.12 - 3.95 (m, 4H), 3.87 (s, 2H, broad), 1.67 (s, 6H), 1.56 (s, 9H).

IH NMR (400 M Hz, Chloroform-d) δ 8.01 (s, IH), 7.48 (s, IH), 4.44 - 4.39 (m, IH), 4.06 - 3.98 (m, IH), 3.44 - 3.39 (m,-107 536.43 A: 2.85 IH), 3.12 (s, IH, v.broad), 2.96 - 2.91 (m, IH), 2.25 (s, IH),

2.10 - 1.74 (m, 7H), 1.71 - 1.46 (m, 16H), 1.39 (s, 3H), 1.03 (s, 3H), 0.97 (s, 3H).

IH NMR (400 M Hz, Chloroform-d) δ 7.92 - 7.78 (m, IH), 7.79 - 7.66 (m, IH), 7.57 (s, IH), 7.53 - 7.47 (m, IH), 7.44-108 554.33 A: 3.20

(s, IH), 4.05 - 4.00 (m, 2H), 3.93 (s, 2H), 3.88 - 3.83 (m, 2H), 2.25 - 2.20 (m, 2H), 1.68 (s, 6H), 1.56 (s, 9H).

IH NMR (400 M Hz, DMSO-d6) δ 12.29 (s, IH, br), 8.21 - 8.13 (m, IH), 8.07 - 8.00 (m, IH), 7.95 (d, J = 9.0 Hz, IH),-109 593.39 A: 4.52 7.82 (s, IH), 7.76 - 7.67 (m, IH), 7.46 (s, IH), 6.82 (d, J =

9.0 Hz, IH), 4.01 (s, 2H), 3.84 - 3.75 (m, 4H), 2.61 (s, 3H), 2.10 - 2.05 (m, 2H), 1.54 (s, 9H), 1.52 (s, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 12.63 (s, IH), 8.15 (d, J = 8.1 Hz, 2H), 7.78 (s, IH), 7.56 (d, J = 8.2 Hz, 2H), 7.52 (s,-110 556.38 A: 4.01

IH), 3.92 - 3.87 (m, IH), 3.63 - 3.55 (m, 2H), 3.52 (s, IH), 3.47 (s, IH), 3.31 - 3.24 (m, IH), 1.56 - 1.46 (m, 21H).

IH NMR (400 M Hz, DMSO-d6) δ 8.21 - 8.14 (m, IH), 8.07 - 8.00 (m, IH), 7.84 (s, IH), 7.77 - 7.68 (m, IH), 7.71 - 7.62-111 553.66 A: 3.61

(m, IH), 7.56 (s, IH), 3.95 (s, 4H, br), 3.81 (s, 2H, br), 3.18 (d, J = 4.6 Hz, 3H), 1.56 - 1.50 (m, 15H).

IH NMR (400 M Hz, DMSO-d6) δ 8.22 - 8.13 (m, IH), 8.07 - 8.00 (m, IH), 7.84 (s, IH), 7.76 - 7.67 (m, IH), 7.57 (s, IH),-112 567.81 A: 3.81

4.15 (s, 2H), 4.10 - 4.02 (m, 2H), 3.83 - 3.75 (m, 2H), 3.23 (s, 6H), 1.56 (s, 6H), 1.53 (s, 9H). LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

IH NMR (400 M Hz, DMSO-d6) δ 8.22 - 8.13 (m, IH), 8.08 - 8.00 (m, IH), 7.84 (s, IH), 7.77 - 7.67 (m, IH), 7.57 (s, IH),-113 581.81 A: 3.94 7.52 (d, J = 8.4 Hz, IH), 4.48 - 4.37 (m, IH), 4.06 - 3.91 (m,

4H), 3.85 (s, IH), 3.32 (s, 2H), 1.54 (s, 9H), 1.24 - 1.22 (m, 6H), 1.22 - 1.18 (m, 6H).

IH NMR (400 M Hz, DMSO-d6) δ 8.22 - 8.14 (m, IH), 8.08 - 8.01 (m, IH), 7.85 (s, IH), 7.77 - 7.68 (m, IH), 7.58 (s, IH),-114 639.83 A: 4.02

7.53 (s, IH), 4.29 - 3.85 (m, 6H), 3.62 (d, J = 1.4 Hz, 3H), 1.60 - 1.49 (m, 21H).

IH NMR (400 M Hz, DMSO-d6) δ 12.83 (s, IH), 8.21 - 8.13 (m, IH), 8.08 - 8.00 (m, IH), 7.84 (s, IH), 7.76 - 7.67 (m,-115 625.45 A: 3.66

IH), 7.58 (s, IH), 7.43 (s, IH), 4.22 - 3.79 (m, 6H), 1.62 - 1.46 (m, 21H).

IH NMR (400 M Hz, DMSO-d6) δ 12.62 (s, IH), 7.40 (s, IH), 7.16 (s, IH), 3.85 (t, J = 5.7 Hz, 2H), 3.57 (d, J = 10.6 Hz, 2H), 3.50 (s, IH), 3.45 (s, IH), 2.71 (dd, J = 14.1, 10.4 Hz,-116 556.48 A: 4.16

IH), 1.85 - 1.61 (m, 4H), 1.48 (s, 12H), 1.45 (s, 9H), 1.33 (td, J = 13.5, 4.2 Hz, 3H), 1.00 (s, IH), 0.98 (s, 3H), 0.95 (s, 3H).

LC-MS

Compound Method:

M+1 NMR

number Rt Time

(min)

1H NMR (400 M Hz, DMSO-d6) δ 12.80 (s, 1H), 8.21 (dd, J = 11.0, 2.1 Hz, 1H), 8.08 (dd, J = 8.5, 2.1 Hz, 1H), 7.87 (s, 1H), 7.75 (t, J = 8.1 Hz, 1H), 7.59 (s, 1H), 6.39 (s, 1H), 4.11 (t, J = 5.7 Hz, 2H), 3.95 (s, 2H), 3.66 (t, J = 5.5 Hz, 2H), 2.41 (s, 3H), 2.30 (s, 3H), 1.58 (s, 9H), 1.55 (s, 6H).

[3], 1H NMR (400 M Hz, DMSO-d6) δ 12.81 (s, 1H), 8.18 (dd, J = 11.0, 2.1 Hz, 1H), 8.04 (dd, J = 8.4, 2.1 Hz, 1H), 7.84

1-117 593.473 A: 3.81 (s, 1H), 7.72 (t, J = 8.1 Hz, 1H), 7.55 (s, 1H), 6.35 (s, 1H),

4.07 (t, J = 5.5 Hz, 2H), 3.92 (s, 2H), 3.63 (t, J = 5.6 Hz, 2H), 2.38 (s, 3H), 2.27 (s, 3H), 1.55 (s, 9H), 1.52 (s, 6H).

[2], 1H NMR (400 M Hz, DMSO-d6) δ 12.72 (s, 1H), 8.18 (dd, J = 11.1, 2.1 Hz, 1H), 8.04 (d, J = 8.5 Hz, 1H), 7.84 (s, 1H), 7.72 (t, J = 8.1 Hz, 1H), 7.55 (s, 1H), 6.36 (s, 1H), 4.07 (t, J = 5.6 Hz, 2H), 3.92 (s, 2H), 3.63 (t, J = 5.6 Hz, 2H), 2.38 (s, 3H), 2.27 (s, 3H), 1.55 (s, 9H), 1.52 (s, 6H).

1H NMR (400 M Hz, DMSO-d6) δ 7.44 (s, 1H), 7.17 (s, 1H), 6.34 (s, 1H), 4.05 (t, J = 5.6 Hz, 2H), 3.91 (s, 2H), 3.62 (t, J = 5.3 Hz, 2H), 2.72 (ddt, J = 12.6, 8.9, 2.9 Hz, 1H), 2.38 (s,

1-118 575.79 A: 3.85

3H), 2.27 (s, 3H), 1.88 - 1.66 (m, 4H), 1.52 (s, 7H), 1.48 (s, 10H), 1.35 (td, J = 13.2, 4.0 Hz, 3H), 0.99 (s, 3H), 0.97 (s, 3H).

Example 2. Activity Assays.

Example 2.1. Protocol for testing PAR-2 compounds in ANTAGONIST mode using the Ca FLIPRTETRA assay (384 well).

[0335] 23000 HT-29 cells (ATCC# HTB-38 ) are plated in 384-well black, transparent-flat bottom plates and incubated in 25 uL of l%FBS/McCoy's media ON at 37°C under an atmosphere of 5% C0₂. The following day, 25uL FLIPR calcium 5 Molecular Devices assay reagent (Cat: R-8186) is added. Plates are incubated at 37°C and r. t. for 45 min and 15 min, respectively. Compounds dilutions are prepared in 100% DMSO using a Biomek FX and diluted 20-fold in HBSS and then 5.5uL is added directly to the cells by the FLIP ^TETRA (10X dilution) using FLIPR Tetra Pipette tips, black, non-sterile, 384 Molecular Devices (Cat: 9000-0764).

[0336] A first read is performed to determine whether the test compounds on their own can activate Ca²⁺ responses.

[0337] The cells are incubated with the compounds for 30 min at r. t. and then read in ANTAGONIST mode in the same plate by stimulating with 6.2 uL of a fixed concentration of the activator: final concentrations: 8 μΜ SLIGKV, 1 U/mL Thrombin, 3.1 U/mL trypsin, or 0.6 μΜ UTP. Representative results are shown in Table 3, below.

Table 3: in vitro Activity Data

PAR-2_counterscreen HT29_TRYP HT29_KV PAR-2_counterscreen

Compound

ANT_FR384_HT29_TH Ι<Ζ₅₀ (μΜ) IC₅₀ (μΜ) ANT_FR384_HT29_UTP number

ROM Ι(Ζ₅₀ (μΜ) IC₅₀ (μΜ) -15 >2 0.002 0.001 >2 -16 >2 0.001 0.0007 >2 -17 >2 0.0009 0.0009 >2 -18 >2 0.004 0.004 >2 -19 >2 0.003 0.003 >2 -20 >2 0.0006 0.0007 >2 -21 >2 0.012 0.011 >2 -22 >2 0.0002 0.0002 >2 -23 >2 0.0002 0.0002 >2 -24 >2 0.002 0.003 >2 -25 >2 0.015 0.01 >2 -26 >2 0.003 0.004 >2 -27 >2 0.002 0.001 >2 -28 >2 0.012 0.005 >2 -29 >2 0.0001 0.0001 >2 -30 >2 0.002 0.002 >2 -31 >2 0.0004 0.0004 >2 -32 >2 0.006 0.004 >2 -33 >2 0.013 0.011 >2 -34 >2 0.0006 0.0005 >2 -35 >2 0.014 0.006 >2 -36 >2 0.0001 0.0001 >2 PAR-2_counterscreen HT29_TRYP HT29_KV PAR-2_counterscreen

Compound

ANT_FR384_HT29_TH Ι<Ζ₅₀ (μΜ) IC₅₀ (μΜ) ANT_FR384_HT29_UTP number

ROM Ι(Ζ₅₀ (μΜ) IC₅₀ (μΜ) -37 1.867 0.0002 0.0001 1.833 -38 >2 0.0004 0.0003 >2 -39 >2 0.0006 0.0006 >2 -40 >2 0.007 0.005 >2 -41 >2 0.0009 0.001 >2 -42 >2 0.0009 0.0009 >2 -43 >2 0.032 0.031 >2 -44 >2 0.004 0.006 >2 -45 >2 1.17 1.257 >2 -46 >2 0.003 0.004 >2 -47 >2 0.002 0.003 >2 -48 >2 0.0003 0.0003 >2 -49 >2 0.008 0.009 >2 -50 >2 0.0007 0.0008 >2 -51 >2 0.011 0.011 >2 -52 >2 0.0001 0.0001 2.033 -53 2 0.001 0.001 1.933 -54 >2 0.24 0.263 >2 -55 >2 0.004 0.004 >2 -56 >2 0.021 0.025 >2 -57 >2 0.042 0.047 >2 -58 >2 0.062 0.084 >2 PAR-2_counterscreen HT29_TRYP HT29_KV PAR-2_counterscreen

Compound

ANT_FR384_HT29_TH Ι<Ζ₅₀ (μΜ) IC₅₀ (μΜ) ANT_FR384_HT29_UTP number

ROM Ι(Ζ₅₀ (μΜ) IC₅₀ (μΜ) -59 >2 0.051 0.044 >2 -60 >2 0.09 0.063 >2 -61 >2 0.0003 0.0003 >2 -62 >2 0.0006 0.0005 >2 -63 >2 0.019 0.015 >2 -64 >2 0.001 0.0007 >2 -65 >2 0.005 0.003 >2 -66 >2 0.021 0.022 >2 -67 >2 0.0007 0.0005 >2 -68 >2 0.5 0.437 >2 -69 >2 0.002 0.002 >2 -70 >2 0.0007 0.0005 >2 -71 >2 0.387 0.577 >2 -72 >2 0.002 0.002 >2 -73 >2 0.0007 0.0005 >2 -74 >2 0.001 0.001 >2 -75 >2 0.003 0.002 >2 -76 >2 0.002 0.002 >2 -77 >2 0.0002 0.0003 >2 -78 >2 0.002 0.001 >2 -79 >2 0.003 0.001 >2 -80 >2 0.001 0.0009 >2 PAR-2_counterscreen HT29_TRYP HT29_KV PAR-2_counterscreen

Compound

ANT_FR384_HT29_TH Ι<Ζ₅₀ (μΜ) IC₅₀ (μΜ) ANT_FR384_HT29_UTP number

ROM Ι(Ζ₅₀ (μΜ) IC₅₀ (μΜ) -81 >2 0.002 0.001 >2 -82 -83 >2 0.005 0.005 >2 -84 >2 0.03 0.036 >2 -85 >2 0.056 0.033 >2 -86 >2 0.006 0.007 >2 -87 >2 0.016 0.009 >2 -88 >2 0.001 0.001 >2 -89 >2 0.013 0.009 >2 -90 >2 0.0008 0.0009 >2 -91 >2 0.14 0.12 >2 -92 >2 0.0008 0.0006 >2 -93 >2 0.003 0.003 >2 -94 >2 0.0009 0.0005 >2 -95 >2 0.002 0.002 >2 -96 >2 0.65 0.34 >2 -97 >2 0.001 0.0006 >2 -98 >2 0.83 0.44 >2 -99 >2 0.11 0.082 >2 -100 >2 0.013 0.023 >2 -101 >2 0.004 0.005 >2 -102 >2 >2 >2 PA -2_counterscreen HT29_TRYP HT29_KV PAR-2_counterscreen

Compound

ANT_FR384_HT29_TH ₅₀ (μΜ) IC₅₀ (μΜ) ANT_FR384_HT29_UTP number

ROM Ι(Ζ₅₀ (μΜ) IC₅₀ (μΜ)

1-103 >2 0.0007 0.0007 >2

1-104 >2 0.011 0.006 >2

1-105 >2 0.0003 0.0002 >2

1-106 >2 0.032 0.031 >2

1-107 >2 0.093 0.075 >2

1-108 >2 0.187 0.2 >2

1-109 >2 0.007 0.007 >2

1-110 >2 0.0003 0.0002 >2

1-111 >2 0.014 0.015 >2

1-112 >2 0.001 0.001 >2

1-113 >2 0.005 0.004 >2

1-114 >2 0.004 0.003 >2

1-115 >2 0.427 0.447 >2

1-116 1.967 0.0003 0.0002 >2

1-117 >2 0.00009 0.00009 >2

1-118 >2 0.0002 0.0002 >2

Example 2.2. Protocol for testing PAR-2 compounds: in vivo rat pharmacokinetics experiments.

[0338] Compounds delivered by intravenous route were formulated using a solution formulation consisting of 5% N-methylpyrrolidone 10% Vitamin E-TPGS and 85% water (v/v/) or a solution formulation consisting of 25% N-methylpyrrolidone 35% polyethylene glycol 400 and 40% water (v/v/). Intravenous formulations were administered by a bolus injection (over approximately 10 seconds) using a hypodermic needle attaches to a syringe (tail vein). The dose volume was varied from 1 to 2 mL/kg and the dose was varied from 0.5 to 1 mg/kg. Compounds delivered orally were formulated as a solution formulation consisting of 5% N-methylpyrrolidone 10% Vitamin E- TPGS and 85% water (v/v/) or as fine suspension using a cellulose derivative suspending agent with or without the addition of a wetting agent / surfactant. Milling of large particles was performed as needed to reduce particle size using focused electroacoustic milling, homogenization or low energy media milling. A spray-dried solid dispersion was prepared for key compounds using 1:1 combination with cellulose derivatives. Test compounds were administered orally by gavage using a gavage plastic needle attached to a syringe. The dose volume was varied from 5 to 10 mL/kg and the dose was varied from 1 to 600 mg/kg.

[0339] Rats were bled by venipuncture (jugular vein) and the samples were collected into tubes containing K₃EDTA. The tubes were placed on wet ice until processed. The samples were centrifuged for approximately 10 minutes (at approximately 4°C) within approximately 30 minutes of collection to prepare plasma or diluted in citrate buffer (1:3) and stored frozen (approximately -30°C). Thawed plasma or whole blood samples were extracted by protein precipitation using an organic solvent followed by centrifugation. The supernatants were separated by high-performance liquid chromatography coupled to mass spectrometry detection. A calibration curve was prepared using reference standard compounds spiked into either blank plasma or whole blood matrices and extracted similarly to unknown samples. Pharmacokinetic parameters were calculated using non-compartmental analysis. Representative results are shown in Table 4, below.

Table 4: Rat PK Data

Compound

Cl_p (mL/min/kg) T 1/2 (h) Vss F (%) number

1-62 7.54 1.27 0.382

1-61 3.93 4.10 0.869

1-48 27.10 0.593 1.14

1-38 11.10 1.55 1.12

1-37 50.00 0.44 0.639

1-36 39.90 0.227 0.302

1-31 26.70 0.513 0.855

1-23 35.50 0.563 1.39

1-22 26.70 1.91 2.36

1-20 2.58 8.56 1.61 14

1-17 19.10 1.76 2.39

1-11 4.18 6.06 1.72

Example 2.3. in vivo pharmacological evaluation of PA -2 pathway inhibitors: rat carrageenan- induced paw edema model

[0340] Male Sprague Dawley rats were housed in groups in a temperature controlled room and were acclimatized in the animal facility for at least three days prior to use. Experiments were performed during the light phase of the cycle. Animals have food and water ad libitum.

[0341] Peripheral inflammation was induced by intra-plantar administration of ΙΟΟμΙ of a 1% w/volume (in saline) carrageenan solution under isoflurane anesthesia. The degree of swelling was assessed using the plethysmometer at 2-3-4-5-6h after carrageenan injection. Test compound or vehicle (0.5%MC /0.1%SDS) was systemically administered at the indicated doses in a volume of 5mL/kg either 0.5-3 hour before carrageenan challenge. Representative results are shown in Table 5, below. Table 5: Inhibition of carrageenan induced paw edema

[0342] While a number of embodiments of this invention have been described, it is apparent that the basic examples may be altered to provide other embodiments that utilize the compounds, methods, and processes of this invention. Therefore, it will be appreciated that the scope of this invention is to be defined by the appended claims rather than by the specific embodiments that have been represented by way of example herein.

[0343] As used herein, all abbreviations, symbols and conventions are consistent with those used in the contemporary scientific literature. See, e.g., Janet S. Dodd, ed., The ACS Style Guide: A Manual for Authors and Editors, 2nd Ed., Washington, D.C.: American Chemical Society, 1997.

Claims

WHAT IS CLAI M ED IS: 1. A chemical entit which is compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

A is cyclohexyl in which each of R¹ and R² is 4-methyl, or

A is phenyl in which R¹ is at the 4-position and R² is at the 3-position;

n is 1 or 2;

E is -CH₂- or -C(O)-; and

Z is N-X or CH-X³;

X is R⁵, -C(0)R⁵ or -S(0)₂R⁵;

each instance of r independently is 0, 1 or 2;

each of R⁵ and R⁶ independently is -(V)_a-Y; wherein

V is Ci_6 aliphatic wherein up to three carbon units of said Ci_₆ aliphatic are optionally and

each instance of J independently is halogen, -C0₂H, -CN, haloCi_₄alkyl or Ci_₄alkyl, wherein up to one methylene unit of each of said Ci_₄alkyl and haloCi_₄alkyl is optionally replaced with -0-, -NR-, -S- or -C(O)-;

heteroaryl having 1-4 heteroatoms independently selected from oxygen, nitrogen and sulfur; or C_1-6 aliphatic, wherein up to three carbon units of said C_1-6 aliphatic are optionally and independently replaced with -0-, -N -, -S-, -C(0)- or -S(0)₂-; and wherein each of the phenyl, 5-6 membered heteroaryl and the C_1-6aliphatic is optionally and independently substituted with 1-4 substituents independently selected from the group consisting of halogen, -CN,

-OP(0)(OH)₂, -P(0)(R)(OH) and

each R independently is -H or C_H alkyl; and

a is 0 or 1;

provided that the compound of Formula I is not any of the following:

-210-

-211-

-212-

-213-

-216-

-218-

-219-

2. The chemical entity of claim 1, wherein the compound of Formula (I) is a compound of Formula (II):

3. The chemical entit of claim 2, wherein:

R¹ is -F, -CI, -CH₃ or -CF₃, and

2 is -H, -F, -CI, -CH₃ or -CF₃; and

- 222 -

The chemical entit of claim 3, where

The chemical entity of claim 1, wherein the compound of Formula (I) is a compound Formula III):

The chemical entit of claim 5, where

he chemical entity of claim 7, where

The chemical entity of claim 9, where

11. The chemical entity of claim 10, wherein:

E is -CH₂-.

12. The chemical entity of claim 10, wherein:

E is -C(O)-.

13. The chemical entity of claim 1, wherein the compound of Formula (I) is a compound of

Formula VI):

wherein:

X⁴ is N;

X⁵ is N or CH; and

each of R⁷ , R⁸ and R⁹ independently is selected from -H, halo, Ci_₄ alkyl, Ci_₄ haloalkyl, -OH, -O ^J6, -NH₂, -N HR^J6, -N(R^J6)₂, -C(0)R^J6, -C0₂H, -C(0)OR^J6, -C(0)NH₂, -C(0)NH R^J6, -C(0)N(R^J6)₂, -S(0)₂NH₂, -S(0)₂N HR^J6, -S(0)₂N(R^J6)₂, -C(0)N(R^J6)S(0)₂R^J6, -C(0)N(R^J6)S(0)₂N HR^J6,

provided that at least one of R⁷ , R⁸ and R⁹ is not -H, wherein each instance of R independently is Ci_₃ alkyl.

The chemical entit of claim 13, where

A is

wherein

R¹ is -F, -CI, -CH3 or -CF₃, and

R² is -H, -F, -CI, -CH3 or -CF₃;

R^7' is -H, halo, C_1-4 alkyl, -OH, -O ^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6, -CN or unsubstituted tetrazolyl;

R^8' is -H, halo, C_1-4 alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6, -CN or unsubstituted tetrazolyl; and

R^9' is -H, halo, d_-4 alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6,-CN or unsubstituted tetrazolyl;

provided that at least one of R⁷ , R⁸ and R⁹ is not -H. 15. The chemical entity of claim 14, wherein:

The chemical entity of claim 1, wherein the compound of Formula (I) is a compound Formula VI.A):

wherein:

X⁴ is N;

X⁵ is N or CH; and

each of R⁷, R⁸ and R⁹ independently is selected from halo, C_1-4 alkyl, C_1-4 haloalkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -N(R^J6)₂, -C(0)R^J6, -C0₂H, -C(0)OR^J6, -C(0)NH₂, -C(0)NHR^J6, -C(0)N(R^J6)₂, -S(0)₂NH₂, -S(0)₂NH ^J6, -S(0)₂N(R^J6)₂, -C(0)N(R^J6)S(0)₂R^J6, -C(0)N(R^J6)S(0)₂NHR^J6,

-C(0)N(R^J6)S(0)₂N(R^J6)₂, -CN and tetrazolyl, wherein said tetrazolyl is unsubstituted substituted with R^J6;

lerein each instance of R^J6 independently is C_1-3 alkyl.

The chemical entit of claim 16, wherein:

R¹ is -F, -CI, -CH3 or -CF₃, and

R² is -H, -F, -CI, -CH3 or -CF₃;

R⁷ is halo, C₁₄ alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6, -CN or unsubstituted tetrazolyl; R⁸ is halo, C₁₄ alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6, -CN or unsubstituted tetrazolyl; and

R⁹ is halo, C_1-4 alkyl, -OH, -OR^J6, -NH₂, -NHR^J6, -C0₂H, -C(0)OR^J6,-CN or unsubstituted tetrazolyl.

The chemical entity of claim 17, where

A chemical entity, which is a compound selected from

1-36

1-37

1-38

1-88

1-97

and pharmaceutically acceptable salts thereof.

20. A chemical entity, which is a compound having the structural formula:

or a pharmaceutically acceptable salt thereof.

21. A chemical entity, which is a compound having the structural formula:

or a pharmaceutically acceptable salt thereof.

22. A chemical entity, which is a compound having the structural formula:

a pharmaceutically acceptable salt thereof. A chemical entity, which is a compound having the structural fo

or a pharmaceutically acceptable salt thereof.

24. A pharmaceutical composition comprising the chemical entity of any one of claims 1-23 and a pharmaceutically acceptable carrier, adjuvant, or excipient.

25. A method for treating a PAR-2 mediated disease in a patient comprising administering to the patient the chemical entity of any one of claims 1-23.

26. A method for treating or preventing inflammation or nociception (pain) in a patient

comprising administering to the patient the chemical entity of any one of claims 1-23.

27. A method for treating inflammatory bowel disease, Crohn's disease, irritable bowel

syndrome, ulcerative colitis, asthma, rheumatoid arthritis, osteoarthritis, gingivitis, atopic dermatitis, psoriasis, systemic lupus erythematosus (SLE), scleroderma, interstitial lung disease, polymyositis, periodontitis, vasculitis, Netherton syndrome, atopic dermatitis, dermatomyositis, uveitis, Alzheimer's disease, Parkinson's disease, multiple sclerosis, inflammatory pain, post-operative incision pain, neuropathic pain, fracture pain, osteroporotic fracture pain, gout joint pain, fibrosis, cancer, diet-induced obesity, adipose inflammation or metabolic dysfunction correlating with PAR-2 expression in a patient comprising administering to the patient the chemical entity of any one of claims 1-23.

28. A method of inhibiting proteolytic activation of PAR-2 in a cell comprising administering to a patient or to a biological sample the chemical entity of any one of claims 1-23.

29. A method of inhibiting PAR-2 activity in a cell comprising administering to a patient or to a biological sample the chemical entity of any one of claims 1-23.